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PhytochemistryISSN 1819-3471
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OPEN ACCESS Research Journal of Phytochemistry
ISSN 1819-3471DOI: 10.3923/rjphyto.2017.90.110
Research ArticleAnti-diabetic Activity of Endophytic Fungi, Penicillium Species ofTabebuia argentea; in Silico and Experimental Analysis1Kumar Kalavathi Murugan, 2Chandrappa Chinna Poojari, 3Channabasava Ryavalad,4Ramachandra Yarappa Lakshmikantha, 5Padmalatha Rai Satwadi, 6Ravishankar Rai Vittal and1Govindappa Melappa
1Department of Biotechnology, Dayananda Sagar College of Engineering, Shavige Malleshwara Hills, Kumaraswamy Layout,560 078 Bengaluru, India2Department of Biotechnology, Shridevi Institute of Engineering and Technology, Sira Road, 572 106 Tumkur, Karnataka, India3Bioneeds India Private Limited, Devarahosahally, Sompura Hobli, Nelamangala Taluk, Bangalore Rural District 562111, Karnataka, India4Department of P.G. Studies and Research in Biotechnology and Bioinformatics, Kuvempu University, Jnana Sahyadri,Shankaraghatta 577 451, Shivamogga, Karnataka, India5Academics, School Of Life Sciences, Manipal University, Karnataka, India6Department of Studies in Microbiology, University of Mysore, Manasa Gangothri, 570006 Mysore, India
AbstractBackground and Objective: The plant and microbial phytochemicals possessing many biological activities with less toxic effects. Hence, presentresearch was aimed to identify phytochemicals in Penicillium species extract and their role in diabetic activity. Materials and Methods: Themethanolic extract of endophytic fungi Penicillium species of Tabebuia argentea was used to analyse phytochemical constituents by GasChromatography Mass Spectrometry (GC-MS). The same extract was used to evaluate the in vitro anti-diabetic activity. The phytochemicals profileobtained from GC-MS was used for in silico anti-diabetic activity against 21 different diabetic proteins/enzymes and ADMET(Absorption, Dissolution, Metabolism, Excretion and Toxicity). The analysis of variance was used to determine the significance of differencebetween treatment groups two-way (ANOVA) followed by SPSS2 (MRX version). Results: The methanol extract of Penicillium species consistedof 18 different phytochemicals and they inhibited the activity of "-amylase, $-glucosidase and dipeptidyl peptidase IV at maximum level. Out of18 phytochemicals, the octadecanoic acid methyl ester and 3 phthalates have shown more interaction with all the 21 diabetic proteins/enzymestested. The octadecanoic acid has shown more interaction with 1dhk, 1nu6, 2wy1, 4y14, 3i2m, 3k35, 4j5t and 5td4. The di-isobutyl isophthalate,dioctyl phthalate and bis-2-ethylhexyl phthalate have shown high interaction with 1m1j, 1ogs and 4acd. The overall observation of present studyshowed that octadecanoic acid is responsible for inducing anti-diabetic activity and the compound has the ability to interact with all the diabeticproteins and inactivate their activity. The in silico investigation clearly indicates how tested compounds interact with different diabeticproteins/enzymes, their role was identified and they were non-toxic and non-carcinogens. Conclusion: The Penicillium species represented potentbioactive compounds in their extract and are responsible for significant in vitro and in silico anti-diabetic activity.
Key words: Penicillium species, phytochemicals, anti-diabetic, molecular docking, ADMET
Received: January 17, 2017 Accepted: February 23, 2017 Published: March 15, 2017
Citation: Kumar Kalavathi Murugan, Chandrappa Chinna Poojari, Channabasava Ryavalad, Ramachandra Yarappa Lakshmikantha, Padmalatha Rai Satwadi,Ravishankar Rai Vittal and Govindappa Melappa, 2017. Anti-diabetic activity of endophytic fungi, Penicillium species of Tabebuia argentea; in silico andexperimental analysis. Res. J. Phytochem., 11: 90-110.
Corresponding Author: Govindappa Melappa, Department of Biotechnology, Dayananda Sagar College of Engineering, 560078 Bengaluru, IndiaTel: +01-7338601980 Fax: +91-80-26660789
Copyright: © 2017 Kumar Kalavathi Murugan et al. This is an open access article distributed under the terms of the creative commons attribution License,which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.
Competing Interest: The authors have declared that no competing interest exists.
Data Availability: All relevant data are within the paper and its supporting information files.
Res. J. Phytochem., 11 (2): 90-110, 2017
INTRODUCTION
World is having problems in disease management inrelated to pathogens resistance, usage of same drugs andcost. The scientists are working on finding available drugsin cheaper cost or finding new drugs at less cost. A search fornewer and more effective agents to deal with diseaseproblems is now under way and endophytes are a novelsource of potentially useful medicinal compounds.Endophytes comprise a large but little-explored share offungal diversity1,2.
The endophytes may provide protection and survivalconditions to their host plant by producing a plethora ofsubstances which, once isolated and characterized, may alsohave potential for use in industry, agriculture and medicine3.At present, endophytes are producing biologically importantbioactive compounds using to manage many infectious andnon-infectious diseases.
Endophytic fungi are of biotechnological interest due totheir potential as a source of secondary metabolites that havebeen proven useful for novel drug discovery4.
Endophytic fungi have been shown to produceseveral pharmacologically important compounds such asantimycotics steroid 22-triene-3b-ol5, anticancer cajanol6,podophyllotoxin and kaempferol7, anti-inflammatoryergoflavin8, antioxidant lectin9, insecticidal heptelidic acid10,immunosuppressive sydoxanthone A, B11 and cytotoxicradicicol12.
Plants used in traditional medicine have played a veryimportant role in the search for new bioactive strains ofendophytic fungi, as it is possible that their beneficialcharacteristics are a result of the metabolites produced bytheir endophytic community13,14.
Tabebuia argentea (Bignoniaceae) is an extensive andyellow blossoming tree and have turned out to be a richwellspring of numerous natural mixes, particularly, of phenolicand polyphenolic nature. The plant is able to produceanticancer agent, lapachol, it has the ability to interfere withthe bioactivities of enzymes known as, topoisomerases, agroup of enzymes that are critical for DNA replication in cells15.The antitumor activity of Lapachol may be due to itsinteraction with nucleic acids and the interaction of thenaphthoquinone moiety between base pairs of the DNA helixoccurs with subsequent inhibition of DNA replication and RNA synthesis16. Other biological activities of Lapachol areantimetastatic activity17, anti-microbial and antifungal18,antiviral19, anti-inflammatory20, antiparasitic16, leishmanicidal21
and molluscicidal activity22. Only three reports are available inthe identification of Lapachol producing endophytes ofTabebuia argentea from our lab research22-24.
Some research works believed to producepharmacologically important bioactive compounds, in thiscontext, the aims of the present study were to characterize thephytochemical profile of fungal endophyte, Penicilliumspecies associated with Tabebuia argentea and to detectanti-diabetic activities and in silico prediction.
MATERIALS AND METHODS
Collection and extraction of phytochemicals fromendophytic fungi, Penicillium species: The endophytic fungi,Penicillium species of Tabebuia argentea were collected fromstock culture unit of Department of Biotechnology,Shridevi Institute of Engineering and Technology, Tumakuru,Karnataka, Bengaluru in September, 2016 and grown in250 mL Erlenmeyer flask containing 100 mL of roseBengal-yeast extract-sucrose broth for 2 weeks at 26+2EC withperiodical shaking at 150 rpm. After the incubation period, theculture was separated from the broth and was extracted usingmethanol as organic solvent. Extraction was done using themycelial mat for the metabolites with methanol. Added theequal volume of the solvent to the filtrate, mixed well for10 min and kept for 5 min till the two clear immiscible layersformed. The upper layer of the solvent containing theextracted compounds was separated using separating funnel.Evaporated the solvent and the resultant compound was driedin rotator vacuum evaporator to yield the crude metabolites25.Then, the extract was dissolved with dimethyl sulphoxideat 1 mg mLG1 of concentration and kept at 4EC.
Phytochemical analysis: The preliminary phytochemicalanalysis of the crude extracts of Penicillium species was doneto know alkaloids, flavonoids, tannins, phenols, saponins,terpenoids and carbohydrates using standard methods25,26.
Detection of bioactive compounds by GC-MS analysis: Themethanol crude extract was subjected to GC-MS analysis toidentify the bioactive compounds. The GS-MS analysis of thecrude extract was carried out in a Shimadzu GC-MS-QP 2010Plus fitted with RTX-5 (60 m×0-25 mm×0.25 µm) capillarycolumn in IISc, Bengaluru. The instrument was set to an initialtemperature of 70EC and maintained at this temperature for2 min. At the end of this period, the oven temperature wasrose up to 2800EC, at the rate of an increase of 50EC minG1 andmaintained for 9 min. An injection port as 1 mL miG1. Theionization voltage was 70 eV. The sample was injected as 10:1.Mass spectral scan range was set at 45-450 (m/z). Theidentification of bioactive compounds present in the extractswas performed by comparing the mass spectra with data fromNIST05 (National Institute of Standards and Technology, US)
91
Res. J. Phytochem., 11 (2): 90-110, 2017
Fig. 1: GC-MS analysis showing different phytochemicals identified based on retention time in methanol extract of endophyticfungi Penicillium species
library. The name, molecular weight and structure of thecomponents of the test material were ascertained based onretention time.
Anti-diabetic activity"-glucosidase activity: A 36 µL of phosphate buffer solution,30 mL sample solution with various concentrations (10, 25, 50,100 and 150 µg mLG1) and 17 µL of 4-nitrophenyl-"-D-glycopyranoside (PNPG) substrate as the concentration of5 mM were put in 37EC for 5 min. After 5 min, 17 µL of"-glucosidase solution 0.15 U mLG1 was added to each well toobtain a total volume of 100 mL. The mixture was incubatedfor 15 min, the reaction was spotted by adding 100 µL ofsodium carbonate 200 mM. Absorbance was measured at405 nm using a microplate reader. Each test was repeatedthrice27. The calculation was done based on Elya et al.28.
"-amylase assay: A 250 mL of 500 µg mLG1 extract, 250 µL ofstarch 2.0% (w.v) and 250 µL of 1 U mLG1 "-amylase solutionwas homogeneously mixed into a test tube. After incubatedat 20EC for 3 min, 500 µL of color reagent (dinitrosalicylic acid)was added to stop the enzymatic reaction. The mixture waskept into boiled water and 250 µL of 1 U mLG1 "-amylase wasadded immediately. The mixture was heated up to 15 min.Further, the solution was removed from the heating processand cooked at room temperature (-26+20EC) for 3 min. A4500 µL aqua dest was added to obtain a total volume of6000 µL. The solution was homogenized using a vortex.
The "-amylase activity was determined at 540 nm usingspectrophotometry to measure product absorbance (maltose)which reduces DNS. The produced absorbance was comparedwith a blank. Percent inhibition was calculated using theequation of Elya et al.28.
Dipeptidyl peptidase IV assay: A 25 µL extract was added to50 µL Dipeptidyl peptidase (DPP-IV) (500 µg mLG1). Themixture was incubated at 37EC for 5 min. A 100 µLGly-Pro-P-Nitroanilide (GPPN) (2 mM) was added to the wellscontaining extract and enzyme. Incubation was contained for15 min. The reaction was terminated by adding 25 µLglacial acetic acid (25%). The absorbance was measured at λ = 405 nm29.
In silico antidiabetic activityBioactive compound preparation: Most of the 3D(3 Dimensions) structures of drug molecules identified in themethanol extract of endophytic fungi, Penicillium specieswere downloaded from PubChem Compound section ofNational Center for Biotechnology Information (NCBI)30.Ligands during this process also being checked for Torsioncount to detect currently active bonds with default settings.Importantly, amide bonds were checked and treated asnon-rotatable. Ligands were also utilized to merge non-polarhydrogens. The 2D structures of 18 ligands are illustrated inTable 1 and Fig. 1. The 3D structures of these 18 ligands wereelucidated.
92
Time (min)
Are
a (%
) 8500000
8000000
7500000
7000000
6500000
6000000
5500000
5000000
4500000
4000000
3500000
3000000
2500000
2000000
1500000
1000000
500000
0 6 8 10 12 14 16 18 20 22 24 26 28 30 32
Res. J. Phytochem., 11 (2): 90-110, 2017
93
Table 1: Id
entif
ied ph
ytoc
hem
icals i
n Pe
nicillium
spec
ies e
xtract and
their s
ynon
ymou
s, iden
tified ba
sed on
rete
ntion tim
e in G
C-MS
Peak
No.
Rete
ntion tim
e (m
in)
Extrac
ted ionic pe
aks
Iden
tified co
mpo
und na
me
Syno
nym
s1
16.681
191.1, 57, 206
.1, 1
92.1, 4
1Ph
enol, 2
,4-b
is (1,1-d
imet
hyleth
yl)-
1.Ph
enol, 2
,4-d
i-ter
t-bu
tyl-
2.2,4-
Di-t
ert-bu
tylphe
nol
3.2,4-
di-t-B
utylph
enol
4.1-
Hyd
roxy
-2,4-d
i-ter
t-bu
tylben
zene
5.2,4-
Bis (
1,1-
dim
ethy
leth
yl) p
heno
l6.
2,4-
Bis (
tert-b
utyl) p
heno
l7.
2,4-
tert-b
utylph
enol
8.2,4-
bis (
1,1-
dim
ethy
leth
yl) p
heno
l2
19.469
121, 91, 41, 93, 77
12-A
zabicy
clo [9.2.2] p
entade
ca-1
(13),
1.12
-aza
bicy
clo[
9.2.2]pe
ntad
eca-1(13
),11,14
-trie
n-13
-am
ine
11,14-
trien-
13-ylam
ine
320
.601
164, 208
, 165
, 190
, 78
1H-2
-Ben
zopy
ran-
1-on
e, 3,4-d
ihyd
ro-
1.(3R)
-8-H
ydro
xy-6
-met
hoxy
-3-m
ethy
l-3,4-d
ihyd
ro-1
H-is
ochr
omen
-1-o
ne8-
hydr
oxy-6-
met
hoxy
-3-m
ethy
l-, (R
)-2.
(3R)
-8-H
ydro
xy-6
-mét
hoxy
-3-m
éthy
l-3,4-d
ihyd
ro-1
H-is
ochr
omén
-1-o
ne
3.1H
-2-B
enzo
pyran-
1-on
e, 3,4-d
ihyd
ro-8
-hyd
roxy
-6-m
etho
xy-3
-met
hyl-, (3
R)-
4.1H
-2-B
enzo
pyran-
1-on
e, 3,4-d
ihyd
ro-8
-hyd
roxy
-6-m
etho
xy-3
-met
hyl-, (R
)-5.
Isoc
oum
arin, 3
,4-d
ihyd
ro-8
-hyd
roxy
-6-m
etho
xy-3
-met
hyl-, (R
)-(-)-
6.(3R)
-8-H
ydro
xy-6
-met
hoxy
-3-m
ethy
l-3,4-d
ihyd
ro-1
H-is
ochr
omen
-1-o
ne
7.(R
)-(-)-
6-m
etho
xym
ellein
8.(R
)-6-m
etho
xym
ellein
9.2,4-
Dihyd
ro-8
-hyd
roxy
-6-m
etho
xy-3
-met
hyl-1
H-2
-ben
zopy
ran-
1-on
e4
20.744
148.9, 57, 41, 150
, 104
1,2-
Benz
ened
icarbo
xylic
acid,
1.Ph
thalic acid, diis
obut
yl ester
bis (
2-m
ethy
lpro
pyl) es
ter
2.Diis
obut
yl pht
halate
; Hex
aplas M
/1B
3.Isob
utyl pht
halate
; Palatinol IC
4.Diis
obut
yles
ter k
yseliny fta
love
5.1,2-
Benz
ened
icarbo
xylic
acid, di (2-
met
hylpro
pyl) es
ter
6.1,2-
Benz
ened
icarbo
xylic
acid, 1,2-b
is (2-m
ethy
lpro
pyl) es
ter
7.Bis (
2-m
ethy
lpro
pyl) ph
thalate
8.Isob
utyl-o
-pht
halate
9.di-2
-met
hylpro
pyl p
htha
late
521
.358
74, 8
7, 43, 55, 41
Hex
adec
anoic ac
id, m
ethy
l ester
1.Pa
lmitic ac
id, m
ethy
l ester
2.n-
Hex
adec
anoic ac
id m
ethy
l ester
3.Met
hyl h
exad
ecan
oate
4.Met
hyl n
-hex
adec
anoa
te5.
Met
hyl p
alm
itate
6.Ac
ide he
xade
cano
ique
met
hyl e
ster
622
.387
163, 148
, 70, 181
, 77
Phth
alic acid, m
ethy
l octyl ester
1.1,2-
Benz
ened
icarbo
xylic
acid m
ethy
l octyl ester
2.
1,2-
Benz
ened
icarbo
xylic
acid, octyl m
ethy
l ester
3.Met
hyl o
ctyl pht
halate
723
.027
67, 8
1, 55, 95, 41
10,13-
Octad
ecad
ieno
ic acid, m
ethy
l ester
1.Met
hyl (10
E,13
E)-1
0,13
octad
ecad
ieno
ate
2.Met
hyl (10
E,13
E)-o
ctad
eca-10
,13-
dien
oate
823
.312
74, 8
7, 43, 55, 41
Octad
ecan
oic ac
id, m
ethy
l ester
1.Met
hyl s
tearate
2.Met
hyl o
ctad
ecan
oate
3.Octad
ecan
oic ac
id, m
ethy
l ester
4.St
earic
acid, m
ethy
l ester
5.St
earic
acid m
ethy
l ester
6.Met
hyl n
-octad
ecan
oate
7.n-
Octad
ecan
oic ac
id m
ethy
l ester
Res. J. Phytochem., 11 (2): 90-110, 2017
94
Table 1: Con
tinue
Peak
No.
Rete
ntion tim
e (m
in)
Extrac
ted ionic pe
aks
Iden
tified co
mpo
und na
me
Syno
nym
s9
24.476
87, 1
48.9, 4
1, 45, 43
Unk
nown co
mpo
und
Unk
nown
1025
.925
287.1, 302
.1, 2
09,
Unk
nown co
mpo
und
Unk
nown
105, 165
1126
.054
287.1, 302
.1, 2
09,
Phen
ol, 2
,4-b
is(1-
phen
ylet
hyl)-
1.2,4-
Bis (
1-ph
enylet
hyl) ph
enol
105, 165
2.2,4-
Bis ("-m
ethy
lben
zyl) ph
enol
3.Ph
enol, 2
,4-b
is(1-
phen
ylet
hyl)-
4.
2,4-
Bis (
1-ph
enylet
hyl) ph
enol
1226
.443
148.9, 57, 166
.9,
Bis (
2-et
hylhex
yl) p
htha
late
1.Ph
thalic acid, Bis
(2-e
thylhe
xyl) es
ter
71, 4
32.
Bis (
2-et
hylhex
yl) 1
,2-b
enze
nedica
rbox
ylate
3.Di (et
hylhex
yl) p
htha
late
4.Di (2-
ethy
lhex
yl) p
htha
late
5.Dioctyl pht
halate
6.Octyl pht
halate
; 2-Eth
ylhe
xyl p
htha
late
7.Ph
thalic acid di (2
-eth
ylhe
xyl) es
ter
8.di-is
o-Octyl pht
halate
9.Di (2-
ethy
lhex
yl) o
-pht
halate
10.
Di-s
ec-o
ctyl pht
halate
11.
Di (2-
ethy
lhex
yl) o
rtho
phth
alate
12.
Bis (
2-et
hylhex
yl) o
-pht
halate
13.
1,2-
Benz
ened
icarbo
xylic
acid, Bis
(2-e
thylhe
xyl) es
terB
is (2-e
thylhe
xyl)
este
r pht
halic
acid
14.
1,2-
Benz
ened
icarbo
xylic
acid, 1,2-B
is (2-e
thylhe
xyl) es
ter
1326
.514
287.1, 302
.1, 1
05,
Phen
ol, 2
,4-B
is (1-p
heny
leth
yl)-
1.2,4-
Bis (
1-ph
enylet
hyl) ph
enol
288.1, 209
2.2,4-
Bis ("-m
ethy
lben
zyl) ph
enol
3.Ph
enol, 2
,4-B
is (1-p
heny
leth
yl)-
4.2,4-
Bis (
1-ph
enylet
hyl)p
heno
l 14
26.604
95, 8
1, 91, 55, 107
Phen
ol, 2
,6-B
is (1,1-d
imet
hyleth
yl)
1.
Mes
itol,
-4-[(
4-hy
drox
y-3,5-
dim
ethy
lphe
nyl)
2.alph
a.4-
(3,5-d
i-ter
t-bu
tyl-4
-hyd
roxy
phen
yl)-,
met
hyl]-
3.4-
[(3,5-d
itert-b
utyl-4
-hyd
roxy
phen
yl) m
ethy
l]-2,6-
dim
ethy
lphe
nol
1526
.863
149, 167
, 57.1,
1,2-
Benz
ened
icarbo
xylic
acid,
1.Di-iso
octyl p
htha
late
; Hex
aplas M
/O71
.1, 4
3.1
diiso
octyl e
ster
2.Iso-
octyl p
htha
late
3.Flex
ol plasticizer
diop
4.Ph
thalic acid, Bis
(6-m
ethy
lhep
tyl) es
ter
5.Ph
thalic acid, diis
ooctyl ester
6.Bis (
6-m
ethy
lhep
tyl) ph
thalate
1627
.335
55, 9
5, 81, 41, 43
Nap
htha
lene
, dec
ahyd
ro-1
,8a-dim
ethy
l-1.
4.be
ta.H
,5.alpha
.-Ere
mop
hilane
7-(1-m
ethy
leth
yl)-, [1
R-(1.alpha
.,4a.be
ta.,
2.10
.alpha
.-Ere
mop
hilane
7.be
ta.,8
a.alph
a.)]-
3.7-
Isop
ropy
l-1,8a-dim
ethy
ldec
ahyd
rona
phth
alen
e4.
1,8a
-dim
ethy
l-7-(p
ropa
n-2-
yl) d
ecah
ydro
naph
thalen
e5.
1,8a
-dim
ethy
l-7-p
ropa
n-2-
yl-2
,3,4,4a,5,6,7,8-
octahy
dro-
1H-n
apht
halene
6.Nap
htha
lene
, dec
ahyd
ro-1
,8a-dim
ethy
l-7-(1
-met
hyleth
yl)-
7. [1
R-(1.alpha
.,4a.be
ta.,7
.bet
a.,8a.alph
a.)]-
1728
.066
95, 1
94.1, 8
1, 149
, 55
5-diet
hylam
ino-
2-nitros
ophe
nol
1.5-
(Dieth
ylam
ino)
-2-n
itros
ophe
nol
1828
.519
149, 43, 55, 122
, 95
6-Isop
rope
nyl-4
,8a-dim
ethy
l-4a,5,6,7,8,
1.6-
Isop
rope
nyl-4
,8a-dim
ethy
l-4a,5,6,7,8,8a
-hex
ahyd
ro-2
(1H)-n
apht
haleno
ne8a
-hex
ahyd
ro-1
H-n
apht
halen-
2-on
e2.
4,8a
-dim
ethy
l-6-p
rop-
1-en
-2-yl-1
,4a,5,6,7,8-
hexa
hydr
onap
htha
len-
2-on
e
Res. J. Phytochem., 11 (2): 90-110, 2017
Table 2: List of enzymes selected for docking studies PDB name Name1dhk Structure of porcine pancreatic alpha-amylase1hny The structure of human pancreatic alpha-amylase at 1.8 angstroms resolution1m1j Crystal structure of native chicken fibrinogen with two different bound ligands1nu6 Crystal structure of human Dipeptidyl Peptidase IV (DPP-IV)1ogs Human acid-beta-glucosidase1v4t Crystal structure of human glucokinase1xu7 Crystal Structure of the Interface Open Conformation of Tetrameric 11b-HSD11y7v X-ray structure of human acid-beta-glucosidase covalently bound to conduritol B epoxide2jfe The crystal structure of human cytosolic beta-glucosidase2oox Crystal structure of the adenylate sensor from AMP-activated protein kinase complexed with AMP2p8s Human dipeptidyl peptidase IV/CD26 in complex 2zj3 Isomerase domain of human glucose:fructose-6-phosphate amidotransferase3ctt Crystal complex of N-terminal Human Maltase-Glucoamylase with Casuarine3k35 Crystal Structure of Human SIRT63l2m X-ray Crystallographic Analysis of Pig Pancreatic Alpha-Amylase with Alpha-cyclodextrin3no4 The crystal structure of the alpha-glucosidase (family 31) from Ruminococcus obeum ATCC 291743wy1 Crystal structure of alpha-glucosidase4acd GSK3b in complex with inhibitor4y14 Structure of protein tyrosine phosphatase 1B
Selection of receptors: The receptors were chosen in light oftheir capacity in the pathway of diabetes. The 3D structure of1DHK, 1HNY, 1M1J, 1NU6, 1OGS, 1V4T, 1XU7, 1Y7V, 2JFE, 200X,2ZJ3, 3CTT, 3K35, 3L2M, 3NO4, 3W37, 3WY1, 4ACD, 4J5T, 4Y14and 5TD4. The receptors selected for present study haveappeared in Table 2. The 3D structures of these receptors wereaccessible in their local shape in PDB database. The 3Ddirections of the receptors were obtained from PDBdatabase. To verify the capacity of the model in reproducingexperimental observation with a new ligand, all thesestructures were analyzed again at the binding site.
Docking simulations: The iGEMDOCKv2.1 was employed forbinding affinity measurement between selected ligands andtargeted proteins of diabetes. The content of configure filewas determined as position of receptor file and ligand file.
ADME TEST: ADME/Toxicity parameters compliance wasevaluated by screening through ADMET-SAR, a commercialtool. The ADMET-SAR is system pharmacology or systemchemical biology and toxicology platform designed for theassessment of would be therapeutic indications, off targeteffects and potential toxic end points of natural products. Inthe studied work, this database/tool was used to predict andevaluate the human metabolism compliance, toxicity riskassessment and mode of action by using standardexperimental data.
Statistical analysis: Analysis of variance two- way (ANOVA) ofSPSS2 (Statistical Package for the Social Sciences) (MRXversion) was used to determine the significance of difference
between treatment groups (<0.05). Means between treatmentgroups were compared for significance using Duncan’s newMultiple Range post-test22.
RESULTS AND DISCUSSION
From qualitative phytochemical analysis of Penicilliumspecies methanol extract exhibited potent bioactivecompounds. The Penicillium species have shown thebioactive phytochemicals such as phenols, flavonoids,terpenoids, tannins, carbohydrates, alkaloids and saponins.Similar results were reported by Sharma et al.31 fromPestalotiopsis neglecta and Bhardwaj et al.25 from Penicilliumfrequentans.
The partially purified crude extract of Penicillium specieswas subjected to GC-MS analysis. Total 18 compounds wereidentified based on retention time and area percentage,molecular formula and weight were identified (Table 1, Fig. 1). The highest amount of 1,2-Benzenedicarboxylic acid, diisooctyl ester was noticed in GC-MS as a high peak. Theendophytic fungi, Colletotrichum gloeosporioides ofPhlogacanthus thyrsiflorus have yielded the phenol,2,4-bis(1,1-Dimethyl ethyl), 1-Hexadecane, 1-Hexadecanol,hexadecanoic acid, octadecanoic acid methyl ester and1-nonadecane26. Bis(2-ethylhexyl) phthalate, Pentanoic acid,Melamine, 4H-Pyran-4-one, 2,3-Dihydro-3,5-dihydroxy-6-methyl-, Dodecane, Nonadecane, 5-Hydroxymethylfurfural,1,2,3-Propanetriol, 1-Acetate, Heptose, Triacetin, 2,3-Dihydroxypropanal, 1-Cycloheptene, D-Allose, Pentadecane,1,5-Anhydrohexitol, 3-Deoxy-D-mannoic lactone, Tetradecane,Heneicosane, 4-Oxo-, 1,2-Benzenedicarboxylic acid and
95
Res. J. Phytochem., 11 (2): 90-110, 2017
70
60
50
40
30
20
10
0
Inhi
bitio
n (%
)
-amylase -glucosidase DPPIVEnzyme
Fig. 2: Enzymes inhibition assay of Penicillium species extractThe percent inhibition of "-amylase, "-glucosidase and DPP IV byPenicillium species methanol extract. The values followed by Mean+SEMremained significantly different at p<0.05
Bis (2-ethylhexyl) phthalate31. Papitha et al.32 have identifiedthe similar bioactive compounds from the plant, Tinosporacordifolia. Many microbes have shown that the secondarymetabolites have the ability to bind with active sites andenzymes, receptors and proteins. Phthalic acid, methyl-octyl ester and Bis (2-ethylhexyl) phthalate were identifiedin the extract and have reported as antimicrobialagents26,31,32. Authors have found Phenol, 2,4-bis(1-phenylethyl)-26 and Phenol, 2,6-bis(1,1-dimethylethyl)-4-[(4-hydroxy-3,5-dimethylphenyl)methyl]-33 from endophytic fungi, Colletotrichum gloeosporioides and Fusarium solani ofdifferent plants. The Naphthalene, decahydro-1,8a-dimethyl-7-(1-methylethyl)-, R-(1. alpha.,4a. beta.,7. beta.,8a. alpha.)]- wereidentified from Phoma herbarum34 and Aquilaria sinensis35.The octadecanoic acid methyl ester was identified from fungalendophytes of Ocimum sanctum and exhibiting manybiological activities36. The results confirm that Penicilliumspecies produce important secondary metabolites and whichare exhibiting many biological activities. Comparing to earlierreports, the study supports the evidence that bioactivecompounds produced by fungal endophytes may not beinvolved in the host-endophytic relationship but may alsohave industrial applications. The endophytic fungal species areexploiting for their important bioactive compounds and theyhaving novel medical applications. The results of presentstudy confirms that endophytic fungal species are able toproduce biologically important medicinal bioactivecompounds. Hence, further studies are required to explorethe secondary metabolites of Tabebuia argentea and itsendophytic fungal species and they can be used formanagement of different diseases.
In vitro antidiabetic activity, the extract of Penicilliumspecies has potentially inhibited the activity of "-amylase,"-glucosidase and dipeptidyl peptidase IV listed in Fig. 2. Theobtained result clearly indicates that, the inhibition of
enzymes is concentration dependent. The "-glucosidase wasinhibited more by bioactive compounds of endophytic fungalspecies and the result was less than positive control standarddrug acarbose. Similar results were reported by many scientistusing endophytic extracts against "-glucosidase37-39. The sameextract was inhibited the activity of "-glucosidase atmaximum level compared to standard drug acarbose. Theextract inhibited the activity of DPP-IV and it was lower thanstandard drug diprotin as a positive control. No reports onendophytic fungal extracts showing the inhibitory action ofdipeptidyl peptidase IV. The present study is the first study oninhibition of dipeptidyl peptidase IV using fungal extract.There are some reports say that, the plant extracts have theability to inhibit the activity of dipeptidyl peptidase IV40-42.
The results concluded that, the endophytic fungal extracthas shown potent antidiabetic activity by in silico assay.Molecular docking was performed on 21 different diabetictarget proteins and with all 18 endophytic bioactivecompounds using iGEMDock2.1. The binding interactions ofthese ligands with target proteins were selected on the basisof binding energy or total energy, VDW and hydrogenbonding interaction. These values along with the hydrogenbond forming residues are presented in Table 3. From theanalysis, the pancreatic "-amylase has shown more interactionwith octadecanoic acid methyl ester followed by Di-isooctylphthalate, Bis-2 ethylhexyl phthalate, Phenol 2,6, bis-2hydroxy-5-methyl Benzenedicarboxylic acid. The octadecanoicacid methyl ester binds with the amino acids of humanpancreatic "-amylase followed by Diisooctyl phthalate, 2,4-Bis(1-phenylethyl)phenol, Phenol 2,6, bis-2 hydroxy-5-methyl.Native chicken fibrinogen amino acids more interactwith Diisobutyl isophthalate followed by Methoxymellein,5-diethylamino, Methyl palmitate. Out of 18, 7 bioactivecompounds able to inhibits the Human Dipeptidyl PeptidaseIV by interacting with different amino acids. The octadecanoicacid methyl ester is able to interact with DPP-IV enzymefollowed Diisooctyl phthalate, Phenol 2,6, bis-2 hydroxy-5-methyl, Dimethyl phenol, Bis-2ethylexyl phthalate,Benzenedicarboxylic acid, 2,4-Bis(1-phenylethyl)phenol(Table 3) (Fig. 3).
The Diisooctyl phthalate have showed highest bindingaffinity with human $-glucosidase followed by octadecanoicacid methyl ester, Phenol 2,6-bis-2 hydroxy-5-methyl,Benzenedicarboxylic acid, Diisobutyl isophthalate, Dimethylphenol, Methoxymellein and Bis-2ethylhexyl phthalate. Theoctadecanoic acid methyl ester is able to interact with humanglucokinase enzyme with more binding energy followed byPhenol 2,6-bis-2 hydroxy-5-methyl, Dimethyl phenol,Diisooctyl phthalate and Methyl palmitate.
96
Res. J. Phytochem., 11 (2): 90-110, 2017
97
Table 3: In
silic
o ant
i-diabe
tic activity
of P
enicillium
spec
ies p
hyto
chem
icals
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
2,4-Bis (1-phenylethyl)phenol
1dhk
-80.58
-73.58
-7.00
Gln5, gln5, gln5, th
r6, g
ln7, se
r81h
ny-8
6.38
-86.38
0.00
tyr6
7, ty
r67, glu18
1, ty
r182
, his1
851m
1j-8
0.53
-70.06
-10.47
Pro6
9, se
r70, gly98
, arg
137, ty
r68, ty
r68, ly
s72, gln73
, gln73
1nu6
-86.47
-79.79
-6.69
Glu69
9, gln76
1, his7
54, h
is757
, his7
57, g
lu69
9, asp
729, asp
729, gln73
11o
gs-7
7.03
-70.47
-6.56
Ala1
, pro
3, ala1, arg
2, pro
3, se
r25, phe
26, a
sp27
1v4t
-79.57
-76.57
-2.50
Asp3
63, p
he31
6, his3
17, h
is317
, glu31
9, arg
358, arg
358, pro
359
1xu7
-93.89
-78.21
-15.68
Gly41
, ser
43, s
er43
, lys
44, lys
44, a
la65
, arg
66, h
is120
, his1
20, ile12
11y
7v-8
1.49
-75.60
-5.89
Gln36
2, his3
65, a
rg28
5, le
u314
, asp
315, phe
316, ala31
8, pro
319, gln36
2, his3
652jfe
-78.81
-68.43
-10.37
Arg3
12, p
he22
5, val22
7, his2
50, p
he33
4, tr
p345
2oox
-86.20
-78.15
-8.04
Phe5
29, a
sp25
0, his4
53, a
rg45
7, arg
459, cys
528, phe
529
2zj3
-75.01
-71.51
-3.50
Gly50
8, glu50
4, glu50
4, le
u507
, arg
511, his6
38, s
er63
93c
tt-9
1.30
-85.30
-6.00
Gly25
8, asp
495, asn
100, arg
108, asp
163, le
u463
, tyr49
4, asp
495
3k35
-89.43
-77.64
-11.79
Asp6
1, his6
6, gly67
, thr
55, p
ro60
, asp
61, h
is66, his6
6, ly
s79, ty
r255
3l2m
-82.32
-76.32
-6.00
Gln5, gln5, gln5, gln5, th
r7, s
er8
3no4
-88.10
-85.80
-2.31
Phe3
30, p
ro34
0, gln34
4, ly
s346
, gln34
4, ly
s346
3w37
-79.98
-74.00
-5.98
Gly28
8, arg
103, arg
103, asp
191, ty
r521
, asn
522
3wy1
-83.91
-78.86
-5.05
Ala3
49, h
is348
, his3
48, lys
352, asn
443, asn
443, his5
15, h
is515
, phe
516
4acd
-75.70
-73.20
-2.50
Pro2
94, s
er66
, phe
67, p
he67
, leu
88, g
ly20
24j5t
-81.31
-77.22
-4.09
Asp6
90, a
sp69
0, asp
690, asn
754, asn
757, asn
757, asn
758, ly
s761
4y14
-78.59
-73.66
-4.93
His2
08, g
ln78
, arg
79, a
rg79
, ser
80, p
ro20
6, pro
210
5td4
-90.04
-75.92
-14.12
His3
05, a
la30
7, gly30
8, gly30
9, arg
303, gly30
4, his3
05, g
ly30
6, gly30
8, gly30
9, phe
348
Di methyl
1dhk
-62.81
-55.81
-7.00
Arg4
21, thr
11, p
ro33
2, arg
398, asp
402
1hny
-65.49
-62.08
-3.41
Lys1
78, tyr67
, tyr67
, glu18
1, ty
r182
, his1
851m
1j-7
4.10
-65.60
-8.50
Thr2
1, th
r22, th
r22, pro
20, thr
21, thr
211n
u6-6
1.87
-54.87
-7.00
Arg3
56, a
rg38
2, arg
356, arg
358, se
r360
, ile37
41o
gs-7
0.13
-67.63
-2.50
Asp2
4, arg
2, pro
3, asp
24, s
er25
, phe
26, m
et49
1v4t
-60.65
-54.03
-6.62
Arg3
27, lys
296, ty
r297
, gly29
9, glu30
0, gly32
81x
u7-7
1.17
-64.17
-7.00
Arg2
52, g
ln21
, arg
198, val20
8, se
r209
, ser
209, ile2
10, a
rg25
2, glu25
41y
7v-6
8.00
-65.50
-2.50
Asp2
4, arg
2, pro
3, asp
24, s
er25
, phe
262jfe
69.09
-62.09
-7.00
His1
20, trp
425,ph
e121
, glu16
5, phe
225, glu16
5, phe
225, ty
r309
, trp
345, tr
p417
, glu42
4, tr
p425
2oox
-73.59
-70.09
-3.50
Ala2
76, p
he57
4, asn
269, asn
269, val27
4, phe
296, glu92
, glu92
2zj3
-69.14
-54.82
14.32
Ser4
20, g
ln42
1, se
r422
, ser
422, th
r375
, gln42
1, gln42
13c
tt-6
9.98
-69.98
0.00
Val184
, pro
206, le
u540
, leu
540, tr
p552
3k35
-64.62
-53.57
-11.05
Asp6
1, his6
6, gly67
, pro
60, h
is66, his6
63l2m
-63.12
-59.83
-3.30
Gly11
2, ile4
9, val51
, thr
52, a
la10
8, gly11
23n
o4-7
0.47
-63.47
-7.00
Tyr5
58, g
lu55
9, ty
r558
, glu55
9, arg
565, arg
565, ty
r566
3w37
-74.13
-67.07
-7.06
Gly70
0, ty
r659
, arg
699, asn
758, gly79
1, glu79
2, glu79
23w
y1-6
1.02
-57.52
-3.50
Arg4
56, a
rg45
7, arg
457, his4
59, p
ro46
0, phe
463, phe
463
4acd
-70.32
-66.82
-3.50
Gly26
2, arg
223, se
r261
, gln26
5, arg
223, gln26
54j5t
-71.94
-68.06
-3.89
Tyr6
6, pro
59, p
ro59
, tyr66
, tyr76
8, his7
854y
14-6
3.83
-53.55
-10.28
Glu75
, glu76
, ala77
, met
74, g
lu76
, val24
9, glu25
25t
d4-9
7.29
-54.63
-12.66
Arg1
95, a
sp19
7, his2
99, a
sn30
0, tr
p58, ty
r62, asp
197, asn
300
Res. J. Phytochem., 11 (2): 90-110, 2017
98
Table 3: Con
tinue
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
5-diethylamino
1dhk
-76.41
-44.63
-31.78
Arg1
95, a
sp19
7, glu23
3, his2
99, a
sp30
0, ty
r62, le
u162
, asp
197, glu23
3, asp
300
1hny
-70.90
-51.18
-19.71
Gly30
9, ala31
0, ile3
12, thr
314, arg
303, gly30
4, ala31
0, ile3
121m
1j-8
4.95
-64.89
-20.06
Cys8
1, cys
46, thr
22, s
er48
, gly49
, cys
46, p
ro47
, ser
48, thr
221n
u6-7
5.89
-65.90
-9.98
Pro4
75, v
al55
8, se
r511
, lys
512, ly
s512
, ile52
9, phe
559, arg
560
1ogs
-83.81
-72.13
-11.67
Thr3
0, ty
r40, phe
31, p
he31
, tyr40
, phe
426, le
u493
1v4t
-83.10
-65.81
-17.29
Phe2
3, arg
377, his3
80, p
he23
, gln24
, gln24
, ser
373, th
r376
, thr
376, his3
801x
u7-8
7.06
-62.06
-25.00
Asn1
27, leu
128, phe
129, his1
30, a
sp13
2, ala18
1, se
r125
, asn
127, asn
127, le
u128
, his1
30, h
is135
1y7v
-76.97
-68.72
-8.25
Asp2
4, arg
2, pro
3, asp
24, s
er25
, phe
26, a
rg49
, tyr41
82jfe
-71.39
-42.50
-28.89
Gln17
, his1
20, g
lu37
3, tr
p417
, glu42
4, tr
p425
, phe
121, tr
p345
, trp
417, tr
p425
2zj3
-85.47
-59.43
-26.04
Cys3
73, s
er42
0, gln42
1, se
r422
, ser
422, th
r425
, thr
428, se
r420
, gln42
1, gln42
13c
tt-8
0.28
-54.57
-25.71
Trp3
91, a
sn39
3, asn
393, asp
414, asn
417, val48
7, tr
p391
, gln48
8, his4
89, h
is489
3k35
-85.74
-69.64
-16.10
Gln11
1, his1
31, p
he62
, gln11
1, gln11
1, his1
313l2m
-84.08
-71.69
-12.39
Tyr1
55, a
sp15
9, cys
160, ly
s142
, thr
143, asp
153, ty
r155
, tyr15
5, gln15
6, gln15
63n
o4-7
8.97
-67.83
-11.14
Arg5
8, le
u435
, asn
57, leu
430, le
u430
, asn
431, ly
s433
3w37
-73.27
-57.05
-16.22
Trp2
29, a
sp23
2, asn
496, se
r505
, lys
506, ala23
1, ile2
33, lys
506, ly
s506
3wy1
-76.55
-57.56
-18.99
Asp4
8, arg
456, arg
457, arg
456, arg
457, arg
457, phe
463
4acd
-80.50
-65.63
-14.87
Val208
, glu21
1, asn
213, le
u207
, arg
209, arg
209, gly21
0, asp
233, ty
r234
4j5t
-73.19
-43.32
-29.87
Leu5
3, arg
54, h
is55, phe
56, a
sp61
, phe
56, th5
8, arg
209
4y14
-70.28
-51.19
-19.09
Gly93
, glu13
6, asp
137, asp
137, ile1
34, p
he13
5, phe
135
5td4
-72.48
-50.56
-21.92
Asp2
36, s
er24
4, se
r244
, ser
245, phe
286, val28
7, val28
7Methoxymellein
1dhk
-70.02
-58.22
-11.80
Gly33
4, arg
398, pro
332, gly33
4, arg
398, asp
402
1hny
-79.91
-56.49
-23.41
Ser3
, gln7, gly9, asp
402, pro
4, th
r6, g
ln8, arg
10, a
sp40
21m
1j-9
1.86
-78.96
-12.90
Cys4
6, cys
81, p
ro47
, ser
48, g
ly49
, ser
48, s
er48
, gly49
, cys
811n
u6-8
2.97
-66.11
-16.86
Arg1
25, a
rg12
5, asp
709, asn
710, gly74
1, tr
p124
, arg
124, arg
125, glu20
5, asn
710, asp
739, his7
40,
his7
401o
gs-8
6.90
-72.70
-14.19
Ser3
8, his4
51, s
er45
5, his4
95, p
he31
, phe
31, leu
493, le
u493
, trp
494, his4
951v
4t-7
8.09
-73.17
-4.92
Arg4
22, g
ln24
, gln24
, glu37
2, se
r373
, thr
376
1xu7
-92.10
-75.24
-16.86
Met
93, g
lu94
, thr
122, arg
66, g
lu94
, his1
20, ile12
1, th
r122
, val14
21y
7v-7
9.36
-74.43
-4.92
Ser2
6, pro
3, asp
24, s
er25
, phe
26, m
et49
, glu50
2jfe
-70.56
-61.30
-9.26
Asn3
19, g
lu32
3, gln31
7, glu32
3, gln32
8, gln32
8, asp
329, asp
329, ile3
322o
ox-8
8.79
-71.30
-17.49
Gly24
4, th
r270
, gln27
1, ala95
, thr
270, th
r270
, leu
272, gly27
3, val27
4, phe
296
2zj3
-77.73
-64.47
-13.26
Gln42
1, se
r422
, ser
376, se
r420
, gln42
13c
tt-8
1.85
-63.66
-18.19
Asp6
67, h
is669
, gln67
0, ly
s715
, val66
8, val66
8, his6
69, g
ln67
03k
35-8
5.48
-63.07
-22.41
Asp6
1, phe
62, a
rg63
, gly64
, thr
213, ala51
, asp
61, a
rg63
, ser
214, gln24
03l2m
-79.56
-64.13
-15.43
Tyr6
2, his1
01, a
sp19
7, tr
p58, tr
p59, ty
r62, ty
r62, val16
3, asp
197
3no4
-92.75
-69.81
-22.94
Asp5
44, a
rg56
5, asp
544, ty
r558
, glu55
9, arg
565, ty
r565
, tyr56
6, ty
r566
, tyr56
63w
37-8
5.41
-68.78
-16.63
Asp2
32, a
sn49
6, ala23
1, ile2
33, a
sn49
6, se
r505
, lys
506, ly
s506
3wy1
-91.41
-61.83
-19.58
His3
48, lys
352, gln43
8, th
r445
, thr
445, le
u348
, leu
433, asp
441, ala44
44a
cd-7
4.75
-58.76
-15.99
Arg2
09, g
ly21
0, glu21
1, val20
8, arg
209, arg
209, gly21
0, asp
233, asp
233, asp
233, ty
r234
4j5t
-71.56
-64.20
-7.35
Asn1
29, a
sp20
2, ly
s203
, val20
5, val20
5, tr
p206
, glu43
54y
14-8
1.03
-69.47
-11.56
Arg7
9, his2
08, g
ly20
9, pro
210, arg
79, a
rg79
, ser
80, leu
204, se
r205
, pro
206, pro
206, po2
105t
d4-7
5.7
-62.13
-13.57
Lys2
00, g
lu23
3, ile2
35, tyr15
1, le
u162
, ala19
8, ly
s200
, his2
01, g
lu23
3, ile2
35, ile23
5
Res. J. Phytochem., 11 (2): 90-110, 2017
99
Table 3: Con
tinue
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
16 1dhk
-67.61
-67.61
0As
n393
, trp
396, ly
s457
, his4
911h
ny-5
7.44
-57.44
0Pr
o4, thr
6, th
r6, g
ln8, arg
10, thr
11, a
sp40
21m
1j-7
9.49
-79.49
0Th
r21, th
r22, th
r83, th
r21, th
r22
1nu6
-64.28
-64.28
-0Gly42
4, pro
426, ly
s523
, lys
523, gln58
6, gln58
61o
gs-7
1.54
-71.54
0Al
a1, a
rg2, pro
3, se
r25, phe
26, m
et49
1v4t
-59.71
-59.71
0As
p78, arg
85, a
rg85
, val86
, met
87, h
is105
1xu7
-75.97
-75.97
0Al
a65, arg
66, h
is120
, ile12
1, th
r122
, val14
21y
7v-6
9.79
-69.79
0Ar
g2, p
ro3, se
r25, phe
26, p
he26
, met
492jfe
-60.32
-60.32
0Al
a246
, his2
56, ile35
2, phe
334
2oox
-73.22
-73.22
0Glu92
, ala26
8, asn
269, asn
269, th
r270
, val27
4, phe
296
2zj3
-61.63
-61.63
0Gly61
3, arg
614, arg
614, pro
615, ly
s631
, lys
631, arg
632
3ctt
-69.41
-69.41
0Ar
g29, ty
r46, val77
, phe
78, lu1
60, p
he16
53k
35-6
2.23
-62.23
0Ar
g63, arg
63, g
ly21
2, th
r213
, ser
214
3l2m
-66.12
-66.12
0Tr
p58, tr
p58, ty
r62, ty
r62, val16
33n
o4-7
1.56
-71.56
0Al
a237
, pro
278, phe
289, asn
309, asn
381
3w37
-62.60
-62.60
0Ar
g773
, arg
773, se
r774
, ser
774, gln83
9, arg
840, tr
p841
3wy1
-65.64
-65.64
0As
p510
, leu
511, pro
512, th
r517
, ala51
8, phe
534
4acd
-60.82
-60.82
0His1
73, h
is173
, ser
236, th
r330
4j5t
-59.38
-59.38
0Ly
s203
, gly43
3, glu43
54y
14-6
8.11
-68.11
0Ty
r46, phe
182, arg
221, gln26
25t
d4-5
9.60
-59.60
0Ty
r2, leu
211, le
u211
, lys
227, pro
228, ile2
3018 1d
hk-6
0.12
-54.25
-5.86
Asn3
, leu
237, ala26
0, ly
s261
, asp
21, a
sp21
1hny
-61.74
-58.24
-3.50
Lys1
78, tyr67
, tyr67
, val12
9, glu18
11m
1j-6
7.82
-65.78
-2.04
Gly49
, pro
82, thr
83, c
ys46
, pro
47, s
er48
, thr
221n
u6-6
2.15
-53.65
-8.50
Arg5
96, a
sp67
8, ile3
19, g
ln32
0, asn
321, arg
596, gly57
21o
gs-7
5.11
-64.61
-10.50
Arg2
85, trp
312, tr
p312
, tyr31
3, ty
r313
, phe
316, le
u317
1v4t
-68.03
-56.85
-11.19
Gly29
9, th
r332
, thr
332, ly
s296
, tyr29
7, glu30
0, glu30
0, arg
327, gly32
8, glu33
11x
u7-6
3.10
-56.10
-7.00
Arg2
52, g
ln21
, asn
207, val20
8, se
r209
, arg
252
1y7v
-62.94
-59.09
-3.85
Pro3
, ser
25, p
he26
, met
49, g
lu50
2jfe
-60.63
-60.63
0.00
Phe1
21, tyr30
9, glu37
3, tr
p417
, glu42
4, tr
p425
2oox
-73.54
-60.57
-12.97
Thr2
70, g
ln27
1, le
u272
, asn
269, asn
269, val27
4, phe
296
2zj3
-67.80
-53.62
-14.18
Ser4
20, g
ln42
1, se
r422
, ser
422, gln42
1, gln42
13c
tt-7
2.40
-68.09
-4.32
Met
567, ala28
5, phe
522, phe
535, ala53
6, ala53
73k
35-7
4.24
-60.10
-14.14
Asp6
1, arg
63, g
ly64
, ala51
, phe
62, p
he62
, arg
63, thr
213, se
r214
3l2m
-65.05
-65.05
0.00
Ile45
3, ile4
65, ile47
9, glu48
4, glu48
4, asp
485, asp
485, ile4
883n
o4-6
8.24
-64.74
3.50
Gln54
5, le
u569
, phe
606, phe
606
3w37
-66.99
-60.99
-6.00
Tyr6
59, g
ly70
0, ty
r39, arg
699, arg
699, asn
758
3wy1
-65.17
-63.73
-1.44
Asp5
10, leu
511, pro
512, phe
516, ala51
8, ala52
2, phe
534
4acd
-65.92
-61.63
-4.29
Gly26
2, arg
223, se
r261
, arg
223
4j5t
-61.42
-54.67
-6.75
Tyr6
6, gly38
3, pro
59, p
ro59
, phe
384, ty
r768
, glu78
44y
14-7
1.78
-60.11
-11.67
Cys2
15, a
rg22
1, arg
221, ty
r46, asp
181, phe
182, se
r216
, ala21
7, arg
221
5td4
-59.71
-54.17
-5.55
Asn5
, pro
4, th
r6, g
ln7, gln8, gln8, arg
10
Res. J. Phytochem., 11 (2): 90-110, 2017
100
Table 3: Con
tinue
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
Eremophilane
1dhk
-67.38
-67.38
0Ly
s35, asn
393, tr
p396
, lys
457
1hny
-57.95
-57.95
0Pr
o4, thr
6, th
r6, g
ln8, th
r11, phe
335, asp
402
1m1j
-68.61
-66.11
-2.5
Thr2
1, pro
20, thr
21, thr
83, p
ro20
1nu6
-68.44
-59.97
-8.47
Arg5
96, a
sp67
8, asn
321, arg
596, gly57
21o
gs-7
1.20
-71.20
0Pr
o3, s
er25
, phe
26, a
rg48
, met
49, g
lu50
1v4t
-58.91
-58.91
0As
p78, arg
85, v
al86
, met
87, h
is105
1xu7
-65.72
-65.72
0Gly41
, lys
44, lys
44, a
sn11
9, his1
20, ile12
11y
7v-7
2.24
-72.24
0Pr
o3, s
er25
, phe
26, m
et49
, glu50
2jfe
-60.38
-60.38
0Ph
e179
, phe
225, tr
p345
, glu42
4, asn
426, gln42
72o
ox-7
5.54
-75.54
0As
n269
, asn
269, glu92
, gly92
2zj3
-59.46
-59.46
0Va
l609
, arg
614, arg
614, pro
615, th
r630
, lys
631, ly
s631
, arg
632
3ctt
-70.28
-70.28
0Al
a285
, leu
286, le
u286
, arg
520, his6
45, lys
776, ly
s776
, asp
777
3k35
-72.70
-72.70
0Al
a51, phe
62, a
rg63
, gln11
1, his1
313l2m
-65.08
-65.08
0Tr
p396
, val45
7, ly
s457
, lys
457, his4
91, g
lu49
33n
o4-7
5.39
-75.39
0Pr
o278
, phe
289, asn
306, phe
378, asn
381
3w37
-61.09
-61.09
0Ly
s421
, lys
421, m
et51
3, his5
14, tyr51
53w
y1-6
4.03
-64.03
0As
p510
, leu
511, pro
512, val51
3, phe
516, th
r517
, ala51
8, phe
534
4acd
-65.52
-65.52
0Ar
g223
, gln26
5, arg
223
4j5t
-56.99
-56.99
0Se
r317
, ile31
8, asn
757, asn
757, ty
r760
, lys
761, ly
s761
, glu76
44y
14-6
0.07
-60.07
0Ar
g79, se
r80, se
r80, le
u204
, ser
209, pro
210, pro
210
5td4
-57.55
-57.55
0Tr
p58, tr
p59, tr
p59, ty
r62, ty
r62, asn
300
Benzenedicarboxylic acid
1dhk
-87.56
-73.29
-14.27
Ser2
70, s
er26
, ser
26, trp
269, se
r270
, ser
270, ly
s13, se
r25, se
r26, gln31
1hny
-78.32
-71.32
-7.00
Gln63
, trp
59, tyr62
, gly10
4, ala10
6, le
u165
1m1j
-77.74
-69.95
-7.80
Ser4
8, val79
, trp
34, s
er48
, ser
481n
u6-8
8.83
-84.16
-4.67
Arg3
56, g
lu34
7, arg
356, arg
356, arg
358, se
r360
, ile37
4, ile3
75, ile37
5, se
r376
1ogs
-92.60
-88.09
-4.52
Cys4
, arg
2, pro
3, se
r25, phe
26, a
rg48
, met
49, g
lu50
1v4t
-82.61
-60.96
-21.65
His3
17, h
is317
, ser
360, asp
363, phe
316, his3
17, p
ro35
9, se
360
1xu7
-95.43
-83.56
-11.87
Tyr2
57, a
rg26
9, asn
270, glu25
4, glu25
5, ty
r257
, arg
269, arg
269, asn
270, arg
273, ly
s274
1y7v
-93.40
-86.11
-7.28
Cys4
, arg
2, pro
3, cys
4, pro
6, se
r25, phe
26, g
ly46
, arg
48, a
arg4
8, m
et49
2jfe
-80.27
-76.77
-3.50
Arg3
12, g
lu16
5, phe
225, arg
312, phe
334, tr
p345
2oox
-77.01
-70.03
-6.98
Asp2
50, a
sp25
0, val27
, leu
28, p
ro29
, val16
0, arg
165
2zj3
-78.96
-70.76
-8.20
Ser4
54, h
is462
, arg
342, gly35
4, cys
459, gly46
0, val46
13c
tt-7
7.53
-75.73
-1.80
Ala2
85, a
la50
9, glu51
0, arg
520, phe
535, phe
535, ly
s770
3k35
-78.99
-67.02
-11.97
Trp1
88, trp
186, le
u184
, asp
185, tr
p186
, trp
186
3l2m
-79.05
-73.14
-5.91
Trp3
88, thr
376, th
r377
, trp
388, tr
p388
, gln39
03n
o4-8
6.80
-74.14
-12.66
Ser4
99, s
er49
9, asn
502, se
r499
, asn
502, arg
506, pro
580, gly58
2, ala61
83w
37-8
7.45
-75.78
-11.67
Arg8
40, s
er77
4, th
r775
, gln83
9, arg
840, arg
840, tr
p841
, trp
841
3wy1
-93.65
-77.45
-16.50
His3
48, lys
352, th
r445
, his3
48, g
ln43
8, asn
443, ala44
44a
cd-8
1.76
-77.19
-4.57
Arg2
09, v
al20
8, arg
209, arg
209, gly21
0, glu21
1, asp
233, asp
233, ty
r234
4j5t
-87.48
-70.48
-17.46
Gln30
8, th
r494
, asn
495, arg
304, m
et46
3, th
r494
, asn
495, asn
495
4y14
-86.98
-78.73
-8.25
Arg7
9, se
r80, arg
79, a
rg79
, ser
80, s
er20
5, pro
206, pro
206
5td4
-79.32
-72.56
-6.76
Gln63
, trp
59, tyr62
, gly10
4, ala10
6, le
u165
Res. J. Phytochem., 11 (2): 90-110, 2017
101
Table 3: Con
tinue
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
Bis-2ethylhexyl phthalate
1dhk
-92.46
-83.94
-8.53
Ser2
70, trp
269, tr
p269
, ser
270, th
r23, val24
, ser
25, s
er26
, ser
261h
ny-8
0.56
-80.56
0.00
Tyr6
7, glu18
1, glu18
1, asn
184, asn
184, his1
85, h
is185
, his2
151m
1j-8
3.71
-80.86
-2.85
Lys1
38, g
ln13
4, arg
137, ly
s138
, tyr68
, tyr68
, lys
72, g
ln73
, gln73
1nu6
-89.57
-89.57
0.00
Glu45
2, arg
453, arg
453, pro
475, gly47
6, gly47
6, se
r511
, lys
512, ly
s512
, phe
559
1ogs
-85.12
-76.81
-8.31
Ala1
, asp
27, p
ro3, cys
4, arg
48, m
et49
, glu50
, leu
51, leu
511v
4t-7
7.52
-62.98
-14.54
Asp2
17, h
is218
, his2
18, g
ln21
9, his2
18, h
is218
, gln21
9, arg
403
1xu7
-84.26
-70.97
-13.29
Arg2
52, g
ln21
, asn
162, asn
162, asn
207, asn
207, arg
252
1y7v
-84.09
-76.45
-7.64
Ala1
, ala1, arg
2, pro
3, pro
3, phe
26, p
he26
, asp
27, a
rg26
22jfe
-80.75
-72.56
-8.20
Arg4
45, v
al51
, val51
, tyr42
9, ty
r429
, phe
440, pro
443, pro
443, arg
445
2oox
-80.30
-80.30
0.00
Leu2
72, p
he42
, lys
44, lys
99, lys
99, leu
272, ly
s44
2zj3
-86.64
-81.98
-4.67
His4
62, thr
448, th
r448
, val44
9, his4
62, a
sn46
4, gly46
6, pro
467, pro
467
3ctt
-87.24
-84.23
-3.01
Phe1
09, a
rg10
8, arg
108, phe
109, phe
109, glu11
0, asp
495
3k35
-100
.24
-98.37
-1.87
Phe6
2, his1
31, leu
184, le
u184
, ile21
73l2m
-83.64
-76.19
-7.46
Trp5
9, asp
356, tr
p59, gln63
, val16
3, val35
4, tr
p357
3no4
-85.27
-80.77
-4.50
Leu6
00, g
lu46
, arg
63, a
rg56
3, arg
563, asp
597, ala59
9, le
u600
, leu
600, asn
601
3w37
-74.11
-74.11
0.00
Glu30
1, th
r681
, thr
681, arg
699, his7
55, g
lu75
6, gly75
7, val78
73w
y1-9
3.07
-85.72
-7.35
Pro2
30, lys
225, le
u227
, ala22
9, pro
230, glu23
1, asn
301, ty
r389
, arg
400
4acd
-101
.80
-93.48
-8.31
Asn3
01, g
ly22
8, ala22
0, pro
230, glu23
1, phe
297, asn
301, m
et30
2, asp
333, asp
333, val33
44j5t
-96.06
-84.50
-11.56
Asn4
71, h
is804
, his8
05, a
sn47
1, asn
471, arg
801, his8
03, h
is804
, his8
05, h
is806
, his8
06, h
is807
4y14
-80.31
-85.33
-4.98
Ser8
0, ly
s73, glu75
, ser
80, g
ln10
2, pro
206, his2
08, g
ly20
9, pro
210, pro
210
5td4
-82.53
-81.66
-0.88
Gln30
2, arg
303, gly30
4, arg
346, phe
348, asn
352, asp
353
Diisobutyl isophathalate
1dhk
-82.76
-74.75
-8.01
Val135
, ser
197, asn
65, a
sn65
, phe
66, g
lu13
4, glu13
41h
ny-7
2.19
-58.01
-14.19
Ser1
32, a
sp13
5, ly
s172
, pro
130, ty
r131
, tyr13
1, tr
p134
, asp
135, ly
s172
, tyr17
41m
1j-9
4.06
-90.24
-3.82
Ser4
8, se
r48, se
r48, gly49
, val79
, pro
47, s
er48
, ser
48, g
ly49
1nu6
-84.06
-73.06
-11.00
Ser349
, thr
351, as
o588
, his5
92, m
et34
8, se
r349
, thr351
, ser37
6, se
r376
, glu37
8, glu37
9, gly38
0, as
p588
1ogs
-89.69
-82.50
-7.19
Cys4
, arg
2, pro
3, cys
4, asp
24, s
er25
, phe
26, a
rg48
, glu50
1v4t
-82.94
-71.57
-11.37
Cys2
30, g
ly25
8, gln28
7, gly22
9, asn
231, glu25
6, glu25
6, gln28
7, glu29
01x
u7-8
2.55
-68.30
-14.25
Lys1
74, tyr25
7, asn
270, arg
273, ly
s174
, glu17
4, glu25
5, asn
270, arg
273, ly
s274
1y7v
-86.40
-82.90
-3.50
Cys4
, pro
3, cys
4, phe
26, a
rg48
, met
49, g
lu50
, glu50
2jfe
-80.20
-67.20
-13.00
Arg4
32, a
sp43
9, phe
440, glu44
1, gln47
, asp
380, ty
r429
, phe
440, glu44
12o
ox-9
9.07
-89.91
-9.16
Thr270
, gln27
1, ala9
5, va
l570
, phe
574, as
n269
, gln27
1, le
u272
, gly27
3, va
l274
, leu2
74, le
u275
, phe
296
2zj3
-70.76
-59.01
-11.75
Thr4
25, a
la42
6, asp
427, val67
7, cys
426, asp
427, val67
7, glu68
03c
tt-7
7.66
-68.16
-9.50
Asp6
49, a
rg65
3, ty
r636
, arg
649, asp
649, arg
653, pro
676, glu76
73k
35-9
5.59
80.94
-14.64
Arg6
3, arg
63, p
he62
, phe
62, g
ln11
1, hos
131, ile2
173l2m
-76.03
-67.54
-8.50
Arg3
89, a
rg39
2, th
r376
, thr
377, tr
p388
, trp
388, arg
389, arg
392
3no4
-91.75
-79.89
-11.85
Tyr5
25, p
he52
6, ty
r141
, asn
439, asn
439, asp
523, phe
531, gly54
93w
37-8
3.27
-78.98
-4.29
Trp8
41, a
rg77
3, se
r774
, thr
775, gln83
9, gln83
9, arg
840, arg
840, tr
p841
, val84
2, val84
23w
y1-8
8.18
-81.50
-6.68
Arg4
00, tyr65
, phe
166, gly22
8, ala22
9, asp
333, asp
333, ty
r389
, phe
397, arg
400
4acd
-93.31
-86.94
-6.37
Pro2
12, a
sn21
3, le
u207
, val20
8, arg
209, gly21
0, glu21
1, asp
233, asp
233, ty
r234
4j5t
-82.16
-60.60
-21.56
Arg7
99, a
rg80
1, se
r802
, ser
802, his8
03, a
rg72
7, arg
727, arg
799, phe
800, arg
801, se
r802
, his8
034y
14-7
7.97
-65.52
-8.56
Arg3
3, se
r146
, glu14
7, asp
148, asp
148, val15
5, val15
5, ly
s197
5td4
-82.58
-74.79
-7.78
Asp3
53, a
sp35
6, arg
303, gly30
4, phe
348, asn
252, asn
252, asp
353
Res. J. Phytochem., 11 (2): 90-110, 2017
102
Table 3: Con
tinue
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
Diisooctyl phthalate
1dhk
-110
.85
-103
.50
-7.33
Ser2
70, s
er26
, trp
269, tr
p269
, ser
270, gly30
9, th
r23, se
r25, gln25
, asn
351h
ny-9
0.84
-87.34
-3.50
Lys2
00, tyr15
1, le
u162
, lys
200, his2
01, g
lu23
3, ile2
35, ile23
5, glu24
01m
1j-8
8.54
-76.09
-12.45
Asn2
66, g
ly26
8, arg
419, asn
266, asn
266, phe
267, gly26
8, ty
r382
, lys
400, gly40
31n
u6-1
03.65
-97.92
-5.73
Asp5
88, g
lu34
7, m
et34
8, se
r349
, thr
351, ile3
75, s
er37
6, se
r376
, asp
588
1ogs
-106
.31
-100
.60
-5.73
Pro3
, cys
4, arg
2, pro
3, cys
4, asp
24, p
he26
, arg
48, m
et49
, glu50
1v4t
-87.15
-78.83
-8.32
Val101
, lys
102, gly94
, glu95
, ser
100, val10
1, ly
s102
, lys
458, ly
s459
, cys
461
1xu7
-91.13
-83.62
-7.62
Asn2
70, lys
174, glu25
5, ty
r257
, arg
269, arg
269, asn
270, pro
271, arg
273
1y7v
-87.52
-84.96
-2.56
His3
65, trp
312, pro
319, his3
65, thr
369, arg
463
2jfe
-75.18
-70.18
-5.18
Gln42
7, m
et17
8, tr
p345
, glu42
4, asn
426, asn
426, gln42
72o
ox-8
4.69
-70.56
-14.12
Asn2
30, a
rg26
0, m
et20
0, asn
230, gly25
4, le
u257
, leu
257, le
u258
, arg
260, gly26
62z
j3-8
3.86
-78.70
-5.16
His5
96, a
rg51
1, arg
594, his5
96, tyr59
8, se
r639
, val64
0, asp
641
3ctt
-82.00
-79.81
-2.19
Ser6
64, a
la69
3, ty
r694
, tyr69
4, lts7
15, lys
715, val71
63k
35-1
03.84
-99.57
-5.27
His1
31, p
he62
, arg
63, h
is131
, met
155, le
u184
, asp
185, tr
p186
, trp
186, ile2
173l2m
-99.40
-90.39
-9.01
Lys2
00, h
is201
, trp
59, tyr15
1, val16
3, ly
s200
, his2
01, g
lu23
3, ile2
35, g
lu24
03n
o4-9
0.52
-84.02
-6.50
Arg1
53, g
ly15
2, arg
153, ty
r155
, arg
412, asp
522
3w37
-98.38
-89.21
-9.17
Arg1
02, a
rg10
2, tr
p104
, glu10
5, ile1
06, p
ro10
7, pro
107, arg
113, gly51
63w
y1-1
01.62
-86.61
-15.01
Ala5
14, h
is515
, phe
516, le
u511
, pro
512, pro
512, val51
3, ala51
4, his5
15, a
la51
84a
cd-8
5.92
-81.06
-4.86
Thr2
32, v
al20
8, arg
209, gly21
0, pro
212, pro
212, th
r232
, asp
233, asn
287
4j5t
-92.88
-85.88
-7.00
His8
03, ile73
4, phe
800, arg
801, his8
03, h
is804
, his8
06, h
is807
4y14
-90.64
-70.18
-20.45
Arg2
4, arg
254, gln26
2, ty
r20, arg
24, a
la27
, arg
254, gly25
95t
d4-9
7.31
-81.07
-16.24
Lys3
22, a
rg38
9, gln39
0, ly
s322
, arg
343, tr
p388
, trp
388, arg
389, glu48
4Dimethyl phenol
1dhk
-58.96
-53.96
-5.00
Ser2
70, v
al24
, trp
269, se
r25, gln31
1hny
-62.16
-56.16
-6.00
Val129
, lys
178, ty
r67, ty
r67, glu18
1, glu18
1, ty
r182
, tyr18
2, his1
851m
1j-7
0.66
-62.16
-8.50
Cys4
6, cys
81, thr
21, thr
22, p
ro82
, thr
831n
u6-9
2.88
-89.37
-3.51
Thr3
65, thr
365, le
u366
, asn
369, ty
r388
, thr
411, se
r412
1ogs
-86.13
-80.82
-5.31
Leu5
1, cys
4, phe
26, a
rg48
, glu50
, leu
51, leu
51, a
rg21
11v
4t-9
0.13
-84.63
-5.50
Arg4
22, g
ln22
, gln24
, leu
25, g
ln26
, glu27
, arg
369, glu37
2, th
r376
, his3
801x
u7-6
5.53
-56.03
-9.50
Thr9
2, m
et93
, glu94
, arg
66, h
is120
, ile12
11y
7v-6
4.34
-55.51
-8.83
Arg4
96, g
ln49
7, asp
453, asp
453, gly45
4, his4
95, g
ln49
7, gln49
72jfe
-95.45
-90.85
-4.60
Gln31
7, asn
319, ly
s321
, glu32
3, ile3
26, g
ln32
8, asn
329, asn
329
2oox
-71.94
-59.10
-12.85
Asp2
45, tyr24
6, se
r247
, asn
269, asn
269
2zj3
-60.86
-47.32
-13.55
Gln42
1, se
r422
, ser
422, th
r425
, gln42
1, gln42
13c
tt-6
9.62
-56.55
-13.07
Arg5
20, thr
775, th
r778
, leu
286, le
u286
, arg
520, his6
45, lys
776, ly
s776
, asp
777
3k35
-71.66
-63.39
-8.27
Asp6
1, gly64
, gln24
0, asp
61, p
he62
, arg
63, thr
213, se
r214
, ser
214
3l2m
-62.12
-52.92
-9.20
Ser1
99, lys
200, ly
s200
, his2
01, g
lu23
3, ile2
353n
o4-6
8.57
-58.49
-10.07
Asn5
61, a
sn61
, glu54
3, tr
p425
, gln56
0, gln56
0, arg
58, a
rg58
, gly59
3w37
-63.25
-56.54
-6.71
Gly70
0, ty
r659
, arg
699, gly79
1, glu79
23w
y1-6
5.32
-52.84
-12.49
Asn2
41, a
sn24
1, asn
495, gly49
8, asp
439, asp
539, le
u240
, asn
495
4acd
-62.04
-50.19
-11.85
Phe2
93, g
ln29
5, arg
92, a
rg27
8, ly
s292
,lys2
92, p
he29
3, phe
293
4j5t
-55.87
-49.64
-6.23
Gln35
8, glu36
1, glu36
1, ile3
62, ile36
2, gly55
64y
14-6
6.05
-63.55
-2.50
Tyr4
6, ty
r46, asp
181, phe
182, ala21
7, arg
221
5td4
-61.84
-49.47
-12.37
Arg3
98, a
rg42
1, arg
398, asp
402
Res. J. Phytochem., 11 (2): 90-110, 2017
103
Table 3: Con
tinue
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
Methyl palmitate
1dhk
-81.60
-81.60
0.00
Ala2
60, lys
261, tr
p269
, ser
25, s
er26
, gln31
1hny
-67.45
-58.22
-9.22
Arg3
92, trp
388, tr
p388
, arg
389
1m1j
-90.30
-86.80
-3.50
Gly84
, gly49
, cys
81, thr
83, p
ro47
, ser
481n
u6-7
0.74
-61.74
-9.07
Asp1
92, tyr19
5, gln12
3, asn
151, tr
p168
, glu19
1, ty
r195
1ogs
-82.90
-80.40
-2.50
Tyr4
0, cys
4, phe
26, a
sp27
, tyr40
, arg
48, g
lu50
, leu
511v
4t-8
5.13
-75.38
-9.75
Gly25
8, ala25
9, gln28
7, gly22
7, gly22
9, cys
230, asn
231, glu25
6, gln28
7, glu29
0, gly41
01x
u7-7
3.80
-65.42
-8.38
Ser1
70, lys
187, se
r169
, tyr17
7, le
u215
, gly21
6, ty
r280
1y7v
-84.54
-78.62
-5.92
Asp2
7, ty
r40, cys
4, phe
26, a
sp27
, arg
48, g
lu59
, leu
512jfe
-77.06
-64.06
-13.00
Arg4
32, a
sp43
9, phe
440, glu44
1, gln47
, gln47
, tyr42
9, phe
440, phe
440, glu44
12o
ox-8
2.81
-79.45
-3.36
His4
53, g
lu92
, asp
98, a
sp98
, lys
992z
j3-6
6.99
-56.21
-10.78
Thr4
46, thr
446, val46
1, gly44
4, se
r454
, ser
454, gly46
0, val46
13c
tt-7
4.10
-69.22
-4.87
Ile52
3, ala28
5, phe
522, ala53
6, ala53
7, m
et56
73k
35-7
0.97
-63.76
-7.20
Asp6
1, gln24
0, asp
61, p
he62
, arg
63, thr
213, se
r214
, ser
214
3l2m
-64.80
-59.15
-5.65
Lys1
42,ly
s142
, asp
153, ty
r155
, tyr15
5, gln15
6, gln15
63n
o4-7
2.24
-63.37
-8.87
Tyr2
00, g
ln20
2, th
r208
, tyr20
0, phe
204, ly
s205
, asp
206
3w37
-64.92
-55.76
-9.17
Val842
, ser
774, arg
840, arg
840, tr
p841
3wy1
-63.10
-50.29
-12.81
Lys3
98, a
sp40
1, gly40
2, glu37
7, val38
0, ly
s398
, lys
398
4acd
-72.60
-58.38
-14.22
Gly26
2, ty
r216
, cys
218, arg
223, arg
223, se
r261
, arg
223
4j5t
-68.01
-60.20
-7.80
Tyr2
9, se
r414
, tyr29
, gly37
1, phe
373, glu37
4, glu41
74y
14-7
2.33
-58.82
-13.51
Gly26
2, ty
r216
, cys
218, arg
223, arg
223, se
r261
, arg
223
5td4
-71.66
-61.15
-10.51
Thr3
76, a
rg39
2, tr
p388
, trp
388, arg
389
Methyl stearate
1dhk
-67.60
-67.60
0.00
Asn3
50, g
lu35
2, val35
4, his7
3, ag7
4, gln13
11h
ny-6
9.62
-62.88
-6.74
Asp1
35, lys
172, ty
r118
, asp
125, ty
r131
, asp
135, ty
r174
1m1j
-87.36
-81.21
-6.15
Gly84
, cys
81, p
ro82
, thr
83, lys
32, p
ro47
, ser
481n
u6-8
1.26
-74.76
-6.50
Arg1
25, a
rg12
5, glu20
5, tr
p627
, trp
629, his7
40, h
is740
1ogs
-78.33
-72.17
-6.16
Tyr3
13, a
la31
8, le
u341
, tyr28
4, ty
r244
, gln28
4, ty
r313
, tyr31
3, phe
316, le
u317
1v4t
-83.86
-74.38
-9.48
Gly25
8, ala25
9, gln28
7, gly22
7, gly22
9, cys
230, asn
230, asn
231, glu25
6, glu26
0, gly41
01x
u7-7
9.38
-75.88
-3.50
Lys2
74, a
rg26
9, asn
270, pro
271, pro
271, ly
s274
, arg
269, asn
270, pro
271, ly
s272
1y7v
-84.87
-78.98
-5.89
Asp2
7, ty
r40, cys
4, phe
26, p
he26
, arg
48, g
lu50
, leu
512jfe
-71.78
-71.78
0.00
Val168
, met
172, phe
225, ala24
6, phe
249, his2
50, ile33
2, tr
p345
2oox
-83.07
-80.42
-2.65
His4
53, g
lu92
, asp
98, lys
992z
j3-6
8.87
-68.87
0.00
Leu6
05, a
rg61
4, arg
614, pro
615, th
r630
, arg
632
3ctt
-71.84
-66.63
-5.21
Ilr52
3, ala50
9, arg
520, phe
555, phe
555, ly
s776
3k35
-70.42
-62.63
-7.79
Gln14
5, asp
194, gln14
5, th
r182
, ser
189, asp
194
3l2m
-70.69
-58.66
-12.03
Arg3
92, thr
376, th
r377
, trp
388, tr
p388
3no4
-85.64
-74.54
-11.09
Arg6
3, arg
459, phe
491, arg
63, a
rg11
73w
37-7
8.11
-75.21
-2.90
Arg7
73, a
rg77
3, se
r774
, thr
775, gln83
9, arg
840, tr
p841
, val84
23w
y1-9
5.25
-81.48
-13.77
His1
05, g
ln17
0, asp
202, phe
147, phe
166, ala22
9, asp
333, ty
r389
, arg
400
4acd
-79.58
-74.34
-5.25
Ser2
36, s
er23
6, le
u207
, leu
207, val20
8, arg
209, gly21
0, glu21
1, pro
212, asp
233, th
r235
4j5t
-66.89
-56.00
-10.89
Arg2
09, p
he58
, arg
209, phe
384, ly
s439
4y14
-70.47
-63.54
-6.93
Arg7
9, arg
79, g
ln10
2, gln10
2, pro
206, pro
206, gly20
95t
d4-7
1.86
-68.36
-3.50
Arg2
0, arg
20, tyr52
, arg
72, s
er73
, gly74
, glu78
, ser
112, se
r113
, thr
114
Res. J. Phytochem., 11 (2): 90-110, 2017
104
Table 3: Con
tinue
PDBs
Bind
ing en
ergy
VDW
H-b
ond
Inte
racting am
ino ac
ids
Octadeconic acid methyl ester
1dhk
-117
.35
-93.33
-24.02
Ser5
5, asn
362, asn
362, se
r55, tr
p44, arg
346, tr
p367
, ile35
8, gly35
9, pro
361, asn
362, asp
381
1hny
-102
.76
-83.72
-19.04
Ser3
, asn
5, gln8, th
r84, asn
5, th
r6, g
ln7, gln7, th
r84, asn
88, a
sn22
0, tr
p221
1m1j
-104
.91
-82.36
-22.55
Ser4
47, trp
448, gly1, his2
, his2
, arg
3, tr
p389
, thr
435, th
r435
, asp
436,trp4
48, g
ly1, his2
, his2
, arg
3, arg
3, pro
4, pro
41n
u6-1
16.96
-98.72
-18.25
Tyr3
72, tyr38
6, se
r412
, tyr37
2, ty
r386
, asp
413, asp
413, ty
r414
, leu
436, se
r437
, leu
445, asn
487
1ogs
-102
.49
-86.73
-15.77
Ala4
38, s
er43
9, se
r465
, ser
465, gln44
0, ly
s441
, lys
441, asn
442, asn
442, asp
443, se
r464
, lys
466
1v4t
-105
.32
-82.83
-22.49
Arg6
3, asn
247, glu24
8, le
u47, glu48
, tyr61
, glu67
, glu24
5, glu24
8, glu24
81x
u7-1
07.04
-60.77
-46.27
Gln21
, ser
202, val20
6, asn
207, val20
8, ile2
10, g
ln25
3, glu25
4, asn
24, g
ln21
, arg
198, se
r202
1y7v
-109
.91
-92.24
-17.67
Arg1
31, tyr13
5, th
r138
, asn
146, his4
95, thr
134, asp
137, asp
137, th
r138
, thr
138, pro
139
2jfe
-104
.37
-90.77
-13.60
Gln12
5, asn
196, ly
s199
, asp
129, se
r135
, glu13
6, pro
181, his1
952o
ox-1
05.58
-67.15
-38.42
Trp4
52, leu
241, th
r245
, gln25
1, arg
33, leu
52, a
sn53
, arg
449, tr
p452
, asn
244, asn
244, arg
332z
j3-1
14.57
-78.06
-36.51
Cys3
73, thr
375, se
r376
, gln42
1, se
r422
, ser
473, ala67
4, ly
s675
, leu
404, gln42
1, asp
427, ty
r434
, ser
676, val67
73c
tt-1
06.02
-74.94
-31.08
His1
15, a
rg28
3, arg
283, his6
45, s
er64
6, ala78
0, gln11
7, gln11
7, phe
119, se
r120
, asn
122, asn
122
3k35
-111
.68
-90.75
-20.94
Arg1
24, a
rg19
3, arg
124, gln14
5, ty
r146
, arg
148, arg
162, th
r165
, glu28
1, gln41
, thr
292
3l2m
-113
.67
-101
.80
-11.81
Gly11
2, th
r113
, val50
, thr
52, trp
59, g
ly10
6, ala10
8, val16
33n
o4-1
07.29
-84.87
-22.42
His3
39, ala32
1, lys3
24, g
lu32
5, gly32
8, lys3
32, ala33
1, his3
39, g
ly33
6, his3
39, a
sp27
4, al
a321
, lys3
24, ly
s324
, glu32
5, glu32
53w
37-9
8.61
-69.18
-29.44
Ala2
02, g
ln20
3, ala20
5, his2
06, g
ln21
9, his2
22, thr
739, gln20
3, his2
06,gln21
9, gly69
0, ty
r740
3wy1
-118
.62
-87.37
-31.25
Asp3
44, h
is515
, phe
516, ala52
9, gln53
1, gln43
9, ala34
3, asp
346, ala34
9, ly
s352
, phe
516, ty
r530
, gln63
1, asp
440
4acd
-101
.61
-78.14
-23.47
Pro3
46, v
al34
8, th
r356
, his3
61, thr
356, phe
360, phe
360, pro
372, ala38
24j5t
-113
.49
87.00
-26.49
Glu46
3, glu47
0, gln61
7, asp
724, arg
727, se
r802
, his8
03, h
lu46
3, ly
s669
, lys
669, arg
727, ty
r728
, pro
731, his8
034y
14-1
16.56
-86.31
-30.24
Arg1
05, a
rg10
5, glu17
0, se
r201
, ser
203, his2
08, a
rg10
5, glu16
1, gln16
7, glu16
7, th
r168
, glu17
0, glu17
0, le
u172
, his2
085t
d4-1
17.70
-96.94
-20.76
gly3
6, ly
s322
, trp
388, arg
389, gln39
0, ly
s35, Lys
35, a
sp37
7, tr
p388
, arg
389, ile3
96, a
sp45
6Phenol 2,6-bis-2 hydroxy-5-methyl
1dhk
-88.03
-80.36
-7.67
Asn3
5, le
u237
, lys
257, ala26
0, ly
s261
, trp
269, se
r311
, gly20
, asp
21, thr
23, g
ln31
1hny
-84.58
-65.04
-19.54
Arg3
43, c
ys37
8, asp
381, tr
p382
, thr
377, gly37
9, asp
381, tr
p382
, trp
388
1m1j
-87.64
-77.54
-10.10
Arg5
1, asp
66, v
al79
, ser
48, a
rg51
, asp
66, s
er48
, ser
48, v
al79
1nu6
-101
.11
-87.42
-13.69
Met
348, asp
588, se
r349
, thr
351, se
r376
, ser
376, glu37
8, asp
588, asp
588, ly
s589
, his5
921o
gs-9
9.39
-83.63
-15.75
Ala1
, asp
27, a
tg47
, ala1, arg
2, pro
3, pro
3, cys
4, se
r25, phe
26, a
sp27
, arg
48, m
et49
, glu50
1v4t
-91.98
-83.05
-8.93
Leu2
5, his3
80, g
ln24
, gln24
, leu
25, g
ln26
, glu27
, glu27
, ser
373, th
r376
, thr
376, his3
801x
u7-9
7.11
-85.75
-11.86
Thr1
24, s
er12
6, se
r125
, asn
127, asn
127, asp
132, his1
34, h
is135
, val18
01y
7v-9
7.69
-84.21
-13.47
Ala1
, asp
27, a
rg48
, arg
2, pro
3, pro
3, cys
4, se
r25, phe
26, a
sp27
, met
49, g
lu50
2jfe
-85.31
-70.89
-14.42
Arg2
03, h
is206
, lys
199, arg
203, arg
203, his2
06, m
et29
6, m
et29
62o
ox-8
9.24
-80.42
-8.81
Arg3
3, arg
449, tr
p452
, asn
244, asn
244, th
r245
, arg
33, a
sn53
2zj3
-84.72
-81.61
-3.11
Arg5
11, leu
515, glu51
8, ly
s621
, glu62
2, his6
38, h
is638
, ser
639
3ctt
-99.52
-81.38
-18.14
Ser4
7, se
r139
, pro
158, ty
r46, ly
s48, pro
137, pro
137, se
r155
, gly15
7, pro
158
3k35
-88.16
-78.15
-10.01
Val256
, asp
257, gly52
, thr
55, thr
55, tyr25
5, ty
r255
, val25
6, asp
257
3l2m
-92.32
-87.14
-5.18
Asn5
3, ile4
9, val51
, thr
52, a
sn53
, ala10
7, gly11
0, gly11
23n
o4-8
9.84
-85.97
-3.87
Tyr5
58, tyr55
8, tr
p425
, trp
425, se
r426
, tyr55
8, glu79
, trp
425, tr
p425
, ser
426, ty
r558
3w37
-91.28
-86.40
-4.89
Arg8
53, a
sp78
5, se
r819
, ser
819, gly82
0, le
u851
, leu
851, ly
s852
, arg
853, phe
908
3wy1
-89.26
-81.92
-7.33
Asn4
47, a
sp34
6, pro
437, his4
38, a
sp44
0, asp
441, pro
442, pro
442, asn
447
4acd
-86.78
-77.15
-9.62
Ser6
6, le
u88, se
r66, phe
67, g
ly68
, val87
, val87
, leu
88, leu
88, a
sn95
4j5t
-89.36
-82.92
-6.44
Glu40
2, glu46
3, arg
467, asp
724, arg
727, arg
727, ty
r728
, his8
034y
14-9
1.49
-83.99
-7.50
Tyr4
6, asp
48, g
ln26
2, ty
r46, asp
48, v
al49
, asp
181, phe
182, asg
221, gln26
25t
d4-8
8.83
-80.33
-8.50
Tyr1
51, h
is305
, trp
58, tyr15
1, ly
s200
, his2
01, g
lu23
3, ile2
35, a
sn30
0, his3
05, g
ly30
6
Res. J. Phytochem., 11 (2): 90-110, 2017
Fig. 3(a-r): Octadecanoic acid methyl ester showing interaction with diabetic enzymes and their binding energy, (a) 1dhk(-117.35), (b) 1hny (-102.76), (c) 1nu6 (-116.96), (d) 1v4t (-102.49), (e) 1xu7 (-104.07), (f) 1y7v (-100.91), (g) 2jfe (-104.37),(h) 200x (-105.58), (i) 2zj3 (-114.57), (j) 3ctt (-106.02), (k) 3k35 (-111.68), (l) 3l2m (-113.67), (m) 3no4 (-107.29), (n) 3w37(-98.61), (o) 3wy1 (-118.62), (p) 4y14 (-116.56), (q) 4j5t (-113.49) and (r) 5td4 (-117.7)
105
(a) (b) (c)
(d) (e) (f)
(g) (h) (i)
(j) (k) (l)
(m) (n) (o)
(p) (q) (r)
Res. J. Phytochem., 11 (2): 90-110, 2017
The Human 11beta-hydroxysteroid dehydrogenasetype I have shown more interaction with octadecanoic acidmethyl ester followed by Phenol 2,6-bis-2 hydroxy-5-methyl,Benzenedicarboxylic acid, 2,4-Bis (1-phenylethyl) phenol,Methoxymellein, Diisooctyl phthalate, 5-diethylamino(Table 3, Fig. 3).
The octadecanoic acid methyl ester have shown moreinteraction with human acid-beta-glucosidase followed byPhenol 2,6-bis-2 hydroxy-5-methyl, Benzenedicarboxylic acid,Diisooctyl phthalate, Diisobutyl isophthalate, Methylpalmitate. Human cytosolic $-glucosidase firmly interact withoctadecanoic acid methyl ester with high energy followed byPhenol 2,6-bis-2 hydroxy-5-methyl. The AMP activated proteinkinase was more interact with octadecanoic acid methyl esterfollowed by Diisobutyl isophthalate, Phenol 2,6-bis-2 hydroxy-5-methyl, Methoxymellein, 2,4-Bis (1-phenylethyl) phenol andDiisooctyl phthalate. The fructose-6-phosphateamidotransferase was inhibited by octadecanoic acid methylester at highest binding energy followed by Bis-2ethylhexylphthalate, 5-diethylamino, Phenol 2,6-bis-2 hydroxy-5-methyl.Statin HMG-coa reductase enzyme was inhibited byoctadecanoic acid methyl ester, Phenol 2,6-bis-2 hydroxy-5-methyl at high binding energy followed by 2,4-Bis(1-phenylethyl) phenol, Bis-2ethylhexyl phthalate(Table 3, Fig. 3).
SIRT6 family member, NAD(+)-dependent proteindeacetylases is able to control genomic stability andtranscriptional control of glucose metabolism. Theoctadecanoic acid methyl ester inhibited this enzyme byshowing highest binding energy followed by Diisooctylphthalate, Bis-2ethylhexyl phthalate, Diisobutyl isophthalate,2,4-Bis (1-phenylethyl) phenol, Phenol 2,6-bis-2 hydroxy-5-methyl, 5-diethylamino, Methoxymellein. The pig pancreatic"-amylase was greatly inhibited by octadecanoic acid methylester followed by Diisooctyl phthalate and Phenol 2,6-bis-2hydroxy-5-methyl. The more interaction was betweencreatinine amidohydrolase with octadecanoic acid methylester by their binding energy followed by Methoxymellein,Diisobutyl isophthalate, Diisooctyl phthalate, Phenol 2,6-bis-2 hydroxy-5-methyl, 2,4-Bis(1-phenylethyl)phenol,Benzenedicarboxylic acid, Methyl Stearate, Bis-2ethylexylphthalate (Table 3, Fig. 3).
Sugar beet "-glucosidase was inhibited by octadecanoicacid methyl ester at a maximum level compared withDiisooctyl phthalate, Phenol 2,6-bis-2 hydroxy-5-methyl,Benzenedicarboxylic acid, Methoxymellein. "-glucosidase wasgreatly inhibited its activity by octadecanoic acid methyl
ester followed by Diisooctyl phthalate, Methyl Stearate,Benzenedicarboxylic acid, Bis-2ethylhexyl phthalate,Methoxymellein, Phenol 2,6-bis-2 hydroxy-5-methyl,Diisobutyl isophthalate. The glycogen synthase kinase-3 betawas showed more binding energy when it interact withBis-2ethylhexyl phthalate, octadecanoic acid methyl ester,Diisobutyl isophthalate, Phenol 2,6-bis-2 hydroxy-5-methyl,Diisooctyl phthalate (Table 3, Fig. 3).
Octadecanoic acid methyl ester conjugate proteintyrosine phosphatase 1B exhibits more interaction followed byPhenol 2,6-bis-2 hydroxy-5-methyl, Diisooctyl phthalate,Benzenedicarboxylic acid, octadecanoic acid methyl esterhave showed highest interaction with processing "-Glucosidase I with high binding energy followed by, Bis-2ethylhexyl phthalate, Diisooctyl phthalate, Phenol 2,6-bis-2 hydroxy-5-methyl, Benzenedicarboxylic acid. The human pancreatic "-amylase have firmly interact with octadecanoicacid methyl ester with more binding energy followedDiisooctyl phthalate, Di methyl, 2,4-Bis (1-phenylethyl)phenol, Phenol 2,6-bis-2 hydroxy-5-methyl (Table 3, Fig. 3).
From in silico anti-diabetic activity, the octadecanoicacid methyl ester able to interact with all most all diabeticproteins/enzymes with high biding energy, whereas thediisobutyl isophthalate have conjugated with 1m1j, iso-octylisophthalate on 1ogs and bis-2-ethylhexyl phthalate on 4acdwith more binding energy. The phthalates have expressed asanti-diabetic potentials in vitro and in silico screening43-45
(Table 3, Fig. 4).The octadecanoic acid methyl ester is responsible for
inducing antidiabetic activity reported by Iqbal et al.46 andHashim et al.47 in vitro and in vivo conditions, respectively.Sasikala and Meenak48, Rajkumar et al.49 have reported thatoctadecanoic acid methyl ester was able to inhibit diabeticenzymes in in silico study. In vitro and in silico anti-diabeticactivity of octadecanoic acid methyl ester was carried out byRaajshree and Chitra50. The present study clearly showed thepresence of important phytochemicals which inducedantidiabetic activity in vitro and in vivo conditions.
From online ADME test reveals that, octadecanoic acidmethyl ester, dimethyl phthalate, Diisooctyl phthalate andbis-ethynyl phthalate are non-toxic AMETS test andnon-carcinogens. The results confirm their degradablecharacters except for the octadecanoic acid methyl ester allare readily biodegradable. The online ADMET test for all thecompounds was carried out but the data represented only forabove mentioned four compounds are represented inTable 4.
106
Res. J. Phytochem., 11 (2): 90-110, 2017
107
Table 4: A
DMET
Pre
dicted
pro
file of
the po
tent
phy
toch
emicals o
f Pen
icillium
spec
ies
Octod
econ
ic acid m
ethy
l ester
Diis
obut
yl pth
thalate
Diis
ooctyl pth
thalate
Bis-2-
ethy
lhex
yl pth
thalate
---------------------------------------------
------------------------------------------------
--------------------------------------------------
------------------------------------------------
Prop
erty
Value
Prob
ability
Value
Prob
ability
Value
Prob
ability
Value
Prob
ability
Bloo
d br
ain ba
rrier
BBB-
0.71
09BB
B+0.93
08BB
B+0.92
72BB
B+0.93
83Hum
an In
testinal abs
orpt
ion
HIA
-0.75
86HIA
+0.97
45HIA
+0.95
47HIA
+0.97
97Ca
co-2
-per
mea
ble
CaCo
2-0.77
37Ca
Co2+
0.71
51Ca
Co2+
0.68
10Ca
Co2+
0.70
03P-
glyc
opro
tein-sub
strate
Subs
trate
0.73
89Non
-sub
strate
0.64
65Non
-sub
strate
0.51
17Su
bstrate
0.50
00P-
glyc
opro
tein-in
hibito
r INon
-inhibito
r0.63
58Non
-inhibito
r0.76
84Non
-inhibito
r0.73
75Non
-inhibito
r0.71
74Inhibito
r0.54
34Non
-inhibito
r0.81
62Non
-inhibito
r0.69
07Non
-inhibito
r0.54
66Re
nal o
rgan
ic cation tran
spor
ter
Non
-inhibito
r0.70
64Non
-inhibito
r0.89
14Non
-inhibito
r0.81
89Non
-inhibito
r0.84
46Distribution
Subc
ellular loc
alization
Mito
chon
dria
0.85
77Mito
chon
dria
0.88
97Mito
chon
dria
0.91
07Mito
chon
dria
0.86
62Metabolism
CYP4
50 2C9
subs
trate
Non
-sub
strate
0.77
89Non
-sub
strate
0.82
06Non
-sub
strate
0.83
28Non
-sub
strate
0.84
79CY
P450
2D6 su
bstrate
Non
-sub
strate
0.85
47Non
-sub
strate
0.88
20Non
-sub
strate
0.87
27Non
-sub
strate
0.86
55CY
P450
3A4
subs
trate
Subs
trate
0.52
22Non
-sub
strate
0.60
97Non
-sub
strate
0.50
00Non
-sub
strate
0.58
81CY
P450
1A2
subs
trate
Non
-inhibito
r0.91
91Non
-inhibito
r0.56
66Non
-inhibito
r0.73
92Non
-inhibito
r0.65
39CY
P450
2C9
inhibito
rNon
-inhibito
r0.92
35Non
-inhibito
r0.66
04Non
-inhibito
r0.75
91Non
-inhibito
r0.74
48CY
P450
2D6 inhibito
rNon
-inhibito
r0.92
70Non
-inhibito
r0.92
18Non
-inhibito
r0.89
86Non
-inhibito
r0.84
32CY
P450
2C1
9 inhibito
rNon
-inhibito
r0.87
88Non
-inhibito
r0.79
06Non
-inhibito
r0.72
83Non
-inhibito
r0.63
10CY
P450
3A4
inhibito
rNon
-inhibito
r0.96
59Non
-inhibito
r0.89
83Non
-inhibito
r0.79
13Non
-inhibito
r0.83
09CY
P inhibito
ry pro
misc
uity
Low CYP
inhibito
ry0.94
02Lo
w CYP
inhibito
ry0.70
03Lo
w CYP
inhibito
ry0.81
25Lo
w CYP
Inhibito
ry0.67
09Excretion-toxicity
Hum
an eth
er-a-g
o-Re
late
dW
eak inhibito
r0.88
36W
eak inhibito
ry0.97
38W
eak inhibito
ry0.90
78W
eak inhibito
ry0.90
63ge
ne in
hibitio
nInhibito
r0.52
77Non
-inhibito
r0.96
21Non
-inhibito
r0.80
10Non
-inhibito
r0.77
55AM
ES te
stNon
- AMES
toxic
0.87
54Non
AMES
toxic
0.91
32Non
AMES
toxic
0.91
32Non
AMES
tox
ic0.93
22Ca
rcinog
ens
Non
-carcino
gens
0.97
35Non
-carcino
gens
0.53
91Non
-carcino
gens
0.74
11Non
-carcino
gens
0.71
16Fish
toxicity
High FH
MT
0.80
01High FH
MT
0.98
62High FH
MT
0.99
62High FH
MT
0.99
59Te
trah
umen
a py
rifor
mis
toxicity
High TP
T0.99
57High TP
T0.96
69High TP
T0.99
99High TP
T0.99
99Hon
ey bee
toxicity
High HBT
0.65
29High HBT
0.65
73High HBT
0.56
83High HBT
0.58
16Biod
egrada
tion
Not
read
y0.80
15Re
ady
0.63
48Re
ady
0.53
83Re
ady
0.63
68biod
egrada
ble
biod
egrada
ble
biod
egrada
ble
biod
egrada
ble
Acut
e or
al to
xicity
III0.53
55IV
0.78
36IV
0.78
63IV
0.71
76Ca
rcinog
enicity
(Thr
ee class)
Non
-req
uire
d 0.73
85Non
-req
uire
d0.54
20W
arning
0.50
66W
arning
0.54
34Ab
sorp
tion
Aque
ous s
olub
ility
-1.766
0Lo
gS-4
.597
3Lo
gS-6
.577
6Lo
gS-6
.280
7Lo
gSCa
Co2 Pe
rmab
ility
0.44
624
LogP
app, cm
secG
11.25
21Lo
gPap
p, cm
secG
11.03
64Lo
gPap
p, cm
secG
11.01
28Lo
gPap
p, cm
secG
1
Distribution, metabolism, excretion, toxicity
Rat a
cute
toxicity
2.09
69LD
50, m
ol kgG
11.29
91LD
50, m
ol kgG
11.19
79LD
50, m
ol kgG
11.08
38LD
50, m
ol kgG
1
Fish
toxicity
1.92
40pL
C50, m
g LG
10.31
53pL
C50, m
g LG
1-0
.075
9pL
C50, m
g LG
10.29
41pL
C50, m
g LG
1
Tetrah
ymen
a py
rifor
mis
toxicity
0.84
98pIGC5
0, µg LG
11.02
47pIGC5
0, µg LG
12.11
00pIGC5
0, µg LG
12.10
76pIGC5
0, µg LG
1
Res. J. Phytochem., 11 (2): 90-110, 2017
Fig. 4: Diisobutyl isophthalate, diisooctyl phthalate and bis-2-ethylhexyl phthalate and their conjugated with (a) 1m1j, (b) 1ogsand (c) 4acd of diabetic enzymes with high binding energy
CONCLUSION AND FUTURE RECOMMENDATIONS
Present day, the endophytes are being utilized as a sourceof novel drug compounds for the betterment of humanhealth. The Penicillium species of Tabebuia argentea haveshown medicinally important phytochemicals by GC-MSanalysis. Anti-diabetic, endophytic fungal extract haveinhibited the "-amylase, "-glucosidase, DPP IV activitystrongly. The octadecanoic acid methyl ester and phthalatesare responsible for inhibition of 21 diabetic proteins/ enzymesactively and they exhibited more binding energy. The presentoutcomes would provide alternate methods of naturalproduct drug discovery which could be reliable, economicaland environmentally safe. Using of these fungi, we canproduce a high amount of bioactive compounds withinshort duration in laboratory conditions. In the study usedalmost all proteins or enzymes for in silico assay to know theiractivity on different enzymes and literature reveals thatnobody has tried all these selected proteins for in silicoanti-diabetic activity. Hence, further in vivo studies aresuggested to investigate to isolate and identify purecompounds which are responsible for diabetic activity fromPenicillium species.
SIGNIFICANCE STATEMENTS
The endophytic fungi, Penicillium species of Tabebuiaargentea methanol extract yielded 18 different bioactivecompounds. The same extract significantly reduced theactivity of "-amylase, "-glucosidase and dipeptidylpeptidase IV enzymes in in vitro experiments. The moleculardocking studies help to know inhibitory activity and bindingmode of endophytic fungal phytochemicals with anti-diabetictarget proteins. The octadecanoic acid methyl ester, dimethylphthalate, di-iso-octyl phthalate and bis-ethylhexyl phthalatehave showed the highest binding affinity and good hydrogenbond interactions with active site residues.
ACKNOWLEDGMENTS
The authors are grateful to Visvesvaraya TechnologicalUniversity (VTU), Belagavi, Karnataka, India for providingfinancial support (Ref No. VTU/Aca./2010-11/A-9/11339 dated7 December 2010) with grant number VTU-RG:11339/2010-11for this investigation. We would also like to thanks Dr S Lokesh,DOS in Applied Botany and Biotechnology, University ofMysore, Mysore, India for assistance in identifying endophyticfungi.
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