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Supplementary information-
A strategic approach for direct recovery and stabilization of Fusarium sp. ICT SAC1
cutinase from solid state fermented broth by carrier free cross-linked enzyme
aggregates
Sandeep A. Chaudhari1, and Rekha S. Singhal1 *
1Food Engineering and Technology Department, Institute of Chemical Technology, Matunga,
Mumbai-400019, India
*Corresponding authors:
Prof. Rekha S. Singhal
Total page: 06
Total figures: 07
1
1. FTIR
The FTIR data were recorded with 2.0 cm-1 resolution, within analytical range of 4000 -
400 cm-1, at room temperature (28 ± 2 °C) and total 100 spectral scan were collected. Three
replica spectra were collected in order to assess its reproducibility. Bands were identified by
comparison with earlier published assignments.
Fig S1. pH optima and stability of free and cut-CLEAs under different pH conditions.
6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.00
20
40
60
80
100
120Free Cutinase cut-CLEAs
pH
Rel
ativ
e A
ctiv
ity (%
)
Fig S2. Temperature optima of free and cut-CLEAs at different temperatures.
20 25 30 35 40 45 50 550
20
40
60
80
100
120
Temperature (°C)
Rel
ativ
e A
ctiv
ity (%
)
2
Fig S3. Thermal studies of free cutinase [F] and cut-CLEA[C]; A) thermal stability; B) TG
and DTG curves (––) TG; (- - - -) DTG.
Free CutinaseCut-CLEA
Temperature ( � C)
Wei
ght l
oss (
%)
Der
ivat
ive w
eigh
t los
s (%
)
3
Fig S4. Correlation of log P, dielectric constant and dipole moment of solvents with
cutinase activity.
Fig. S4A
5.6 4.57 3.5 3.15 0.63 0.58 0.34 0.32 -1.3 -0.23
0.28 -0.3 -0.76
0102030405060708090
100
Free Cutinase cut-CLEA
log P values of solvents tested
Rel
ativ
e A
ctiv
ity (%
)
Fig. S4B
46.68 32
.724
.621
.0120
.3319
.9217
.93 17.8 8.1 3.4 2.2
5 2 1.90
102030405060708090
100
Free cutinase cut-CLEA
Dielctric constant (ε) values of solvents tested
Rel
ativ
e A
ctiv
ity (%
)
4
Fig. S4C
46.7 2.88 1.76 1.7 1.69 1.68 1.66 1.66 1.64 1.6 0.45 0.07 00
102030405060708090
100
Free Cutinase cut-CLEA
Dipole moment of solvent
Rel
ativ
e A
ctiv
ity (%
)
Fig S5. Lineweaver-Burk plot (1/S vs. 1/V) of free cutinase and cut-CLEA.
0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 1.20
0.01
0.02
0.03
0.04
0.05
0.06
0.07
0.08
f(x) = 0.0581869745256428 x − 0.0143893884253717R² = 0.946389481357465
f(x) = 0.0821690478871904 x − 0.0186016820007986R² = 0.984385561872146
f(x) = 0.046575576187062 x − 0.011646347399791R² = 0.944382513013829
Free cutinase Linear (Free cutinase)BSA cut-CLEA Linear (BSA cut-CLEA)
1/S (mM)
1/V
(µm
ol/m
in)
5