Functional Properties of Lupinus angustifolius Seed …downloads.hindawi.com/journals/jfq/2017/8675814.pdfSeed Protein Isolates AntonioHilarioLara-Rivera, 1 PedroGarcía-Alamilla,

Research ArticleFunctional Properties of Lupinus angustifoliusSeed Protein Isolates

Antonio Hilario Lara-Rivera1 Pedro Garciacutea-Alamilla2 Laura Mercedes Lagunes-Gaacutelvez2

Ramoacuten Rodriacuteguez Macias1 Pedro M Garciacutea Loacutepez1 and Juan Francisco Zamora Natera1

1Centro Universitario de Ciencias Biologicas y Agropecuarias Departamento de Botanica y Zoologıa Universidad de Guadalajara45110 Zapopan JAL Mexico2Division Academica de Ciencias Agropecuarias Universidad Juarez Autonoma de Tabasco Carretera Villahermosa-Teapa Km 25Ra La Huasteca 2da Secc 86298 Centro TAB Mexico

Correspondence should be addressed to Pedro Garcıa-Alamilla pedrogarciaaujatmx

Received 13 June 2017 Revised 7 October 2017 Accepted 13 November 2017 Published 7 December 2017

Academic Editor Susana Fiszman

Copyright copy 2017 Antonio Hilario Lara-Rivera et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Protein isolates prepared by alkaline solubilization followed by isoelectric precipitation and freeze-drying from six varieties ofLupinus angustifolius (Haags Blaue Sonate Probor Borlu Boregine and Boruta) grown in Mexico were evaluated for functionalproperties nitrogen solubility water-holding capacity (WHC) oil holding capacity (OHC) emulsion activity index (EAI) emulsionstability index (ESI) foaming capacity (FC) foam stability (FS) and gellingminimumconcentration (GMC)Thenitrogen solubilityvalues WHC OHC and FC did not show significant differences between the protein isolates The solubility of the isolates wasminimal at pH of 40 and 50 while the regions of maximum solubility were found at pH of 20 and 100 There were significantdifferences in EAI and ESI depending on the varieties used The isolates of the Boregine and Borlu varieties showed the highestEAI with 293 and 283m2 gminus1 respectively while the lowest index was recorded in the isolate obtained from the Sonate variety(246m2 gminus1) Like solubility these indices also increased at both extremes of pH evaluated both properties were minimal in theisoelectric pH range (40 to 50)

1 Introduction

Legume seeds represent an important and economical sourceof protein for human and animal feeding [1] Its amino acidcomposition is generally adequate however some speciescontain deficient amounts of some amino acids [2] Soybeansworldwide represent themain source of protein neverthelessthe food industry is in the search for new sources of vegetableproteins of good nutritional quality and functional charac-teristics superior or equal to soybean proteins [1 2] Amongthe species most likely to compete with soybean proteins arelegumes of the genus Lupinus [3 4] Historical antecedentsindicate that different cultures of the Mediterranean andSouth America practiced for several years the cultivationof some species of the genus Lupinus to include them intheir diet like Lupinus albus L luteus L angustifolius andL mutabilis [5 6] Due to its high protein content and to

the new varieties with low alkaloid content the cultivation ofthese legumes has spread to other regions of the world for usein animal and human nutrition [7ndash9] In addition Lupinusare more adaptable to regions with low fertility soils andmild or very cold winters compared to soybean cultivation[10] The seeds of these species represent a functional foodingredient because their proteins are related to the preventionof cardiovascular diseases and it had been found that theirproteins (conglutins) reduce blood glucose in hyperglycemicrats [11 12] In this way obtaining protein isolates of Lupinusby isoelectric precipitation is an alternative for the commer-cial production of food ingredients At present the foodindustry has shown a growing interest in protein isolatesdue to their functional properties such as water absorptionoil absorption emulsification foam formation and gelationThese properties are essential to determine the possible usesof these as food ingredients Functional properties provide

HindawiJournal of Food QualityVolume 2017 Article ID 8675814 8 pageshttpsdoiorg10115520178675814

2 Journal of Food Quality

information on how a particular ingredient would behave ina food matrix play a prominent role during the preparationprocessing and storage of food and in turn correlate withsensory properties These properties could be considered asa result of different conformational changes or interactionsbetween food components such as interactions betweenproteins proteins and polysaccharides lipids phenolic com-pounds or phytic acid [13] Furthermore they constitute thefunctional base of diverse products mainly of low proteincontent and high fat content On the other hand legumesare source of beneficial compounds that have a protectiveeffect on the development of various diseases [14] Thereforethe objective of the present research work was to determinethe functional properties of protein isolates of six varietiesobtained from Lupinus angustifolius cultivated in the state ofJalisco Mexico

2 Material and Methods

21 Vegetable Materials Seeds of six varieties of L angus-tifolius (Haags Blaue Sonate Probor Borlu Boregine andBoruta) were used which were provided by the Germancompany Saatzucht Steinach GmbH dedicated to geneticimprovement These seeds were sown and cultivated duringthe fallwinter period of 20142015 under irrigated conditionson agricultural soils of the University Center of Biologicaland Agricultural Sciences of the University of Guadalajara inZapopan Jalisco MexicoThis site is located at the geograph-ical coordinates 20∘4410158404710158401015840N and 103∘3010158404310158401015840W at an altitudeof 1523m It is characterized by a humid temperate climatewith rains in summer with precipitations ranging from 700to 1400mm per year and an annual average temperature of120 to 180∘C with the presence of frost After conventionalland preparation with a moldboard plow and a disk harrowseeding was performed manually under an experimentalrandomized block design with four replicates

22 Proximate Composition and Preparation of Protein Iso-lates After harvesting on the order of 50 kg of seeds onebatch of 1 kg each variety was separately ground with a knifemill (IKA) until flour with a particle size of 2mm wasobtained Every variety was subjected to triplicate a proximalchemical analysis according to the techniques described inAOAC (2000) moisture content (92509) crude fat (92030)crude protein (97909) ash (92303) crude fiber (96209)and carbohydrate content [15] The flour was subjected toa degreasing process by Soxhlet extraction using hexane assolvent [16] The defatted sample (100 g) was used to obtainthe protein isolates

The protein isolate was obtained using the isoelectricprecipitation method [17] A dispersion of 100 g Lminus1 inNa2SO3(0025) was prepared and adjusted to pH of 90

using NaOH (1N) Immediately it was stirred for one hourat room temperature (25∘C) and centrifuged at 10976timesg(HERMLELabortechnikGmbHZ 326Germany) for 10minThe supernatant was acidified to pH of 45 using HCl (1 N)This was subjected to centrifugation at 10976timesg for 10minat 4∘C recovering the precipitate A washing process withdistilled water was applied to the precipitate and NaOH

(1N) was used to bring it to a pH of 7 The precipitate waslyophilized and stored at 4∘C The protein isolates obtainedfrom the seeds of the six varieties were analyzed in triplicatefor all functional properties

23 Functional Properties

231 Solubility Thesolubility of the protein isolates was eval-uated according to the methodology proposed by Ogunwoluet al [18] and Liu et al [19] with slight modifications Eachisolate was prepared in a 01 wv suspension in 10mL ofdistilled water The pH was adjusted in a range of 20ndash100using NaOH (01 N) or HCl (01 N) The suspension wasstirred for 30min on an orbital shaker (Thermo Fisher Sci-entific Multipurpose rotator 2346 China) Subsequently itwas centrifuged for 10min at 10976timesg (HERMLELabortech-nik GmbH Z 326 Germany) The protein content in thesupernatant was determined by the Bradford method Thestandard curve was made from bovine serum albumin (BSA)and the absorbance of the samples was measured at 595 nmon a spectrophotometer (Thermo Fisher Scientific G10S Uv-Vis China)The solubility of the protein was calculated by thefollowing equation

Solubility =protein in supernatant (g)protein in sample (g)

times 100 (1)

232 Water-Holding Capacity (WHC) The WHC was eval-uated by the method of Chau et al [20] with slight modi-fications Samples of protein isolates (05 g) were dispersedin 5mL of distilled water and stirred for 24 h After thistime the centrifugation for 30min at 1474 g (HERMLELabortechnik GmbH Z 326 Germany) was carried out Thevolume of the supernatant wasmeasured after centrifugationThe difference between the initial volume of water and thevolume recovered after centrifugation determines the WHCexpressed as milliliters of water by one gram of sample(mL gminus1)

TheWHC was determined using the following equation

WHC (mLgminus1) = 119881119894 minus 119881119904119882119898

(2)

where119881119894is the initial volume of distilled water (mL)119881

119904is the

volume of the supernatant (mL) and119882119898is the weight of the

sample (g)

233 Oil Holding Capacity (OHC) The OHC was evaluatedby the method of Chau et al [20] with slight modifications05 g of the protein isolates that were obtained was added5mL of canola oil contained in a 50ml centrifuge tubewhich was stirred for 30min by magnetic stirring Then it iscentrifuged at 1474 g (HERMLE Labortechnik GmbH Z 326Germany) for 30 minutes After centrifugation the volumeof the supernatant is measured The difference between theinitial volume of oil and the volume recovered correspondsto the OHC expressed in mL gminus1 of protein isolates

Journal of Food Quality 3

Table 1 Chemical composition of seed flour of varieties of Lupinus angustifolius cultivated in Mexico ( dry basis)

Variety Moisture Protein Ash Fat Crude fiber CarbohydratesHaags Blaue 359b 3233b 350a 440b 805c 4813b

Boregine 365b 2923c 334a 388c 822c 5168a

Borlu 438a 3342b 313a 417b 1006b 4484b

Probor 384b 3661a 325a 420b 986ab 4224b

Sonate 379b 3250b 351a 529a 1249a 4242a

Boruta 368b 2840c 343a 531a 1512a 4406a

Different letters in rows indicate significant differences (119875 lt 005)

The OHC was determined by the following equation

OHC (mLgminus1) = 119881119894 minus 119881119904119882119898

(3)

where 119881119894is the initial volume (mL) 119881

119904is the volume of the

supernatant (mL) and119882119898is the weight of the sample (g)

234 Emulsion Activity Index (EAI) and Emulsion StabilityIndex (ESI) EAI and ESI were determined using the methoddescribed byZheng et al [21] with slightmodifications whichconsisted of mixing 10mg of the protein isolates with 10mLof distilled water together with 333mL of canola oil ThepH of the dispersion was adjusted in a range of 20ndash100by the addition of HCl (01 N) or NaOH (01 N) accordingto the case Subsequently the mixtures were homogenizedon a vortex stirrer (IKA Mixing Model MS1) for 1min at2500 rpm Then a 5mL aliquot of each emulsion formedwas mixed with 5mL of 01 sodium dodecylsulfate Thesamples were analyzed on a spectrophotometer (ThermoFisher Scientific G10S Uv-Vis China) and the absorbancewas read at 500 nm

The EAI was determined by the following equation

EAI (m2gminus1) = 2 times 2303 times 1198600 times DF119888 times 0 times 1 minus 120579

(4)

where 1198600is the zero minute absorbance DF is the dilution

factor (100) 119888 is the initial protein concentration (01 g100mLminus1) 0 is optical path (001m) and 120579 is the fractionof oil (vv) used to form the emulsion (025) After readingthe absorbances in zero time the emulsions were allowed tostand at room temperature (25∘C) for 10 minutes (119860

10) The

absorbance of emulsions was obtained again and the emul-sion stability index (ESI) was calculated with the followingequation

ESI (min) =1198600

1198600minus 11986010

times 10 (5)

235 Foaming Capacity (FC) and Foam Stability (FS) FCand FS were determined following the method describedby Cheung and Chau [22] 05 g of protein isolates wasmixed with 25mL of distilled water adjusted to pH 7 by theaddition of HCl (01 N) or NaOH (01 N) They were thenshaken in a Oster (Sunbeam Mexicana SA de CV ModelBLSTEP7808W-013 Mexico) at the ldquolowestrdquo speed (mode 1

of 10) for 5min The mixture was transferred to a graduatedcylinder (50mL) After 30 seconds the initial foam volumewasmeasured as well as after 5 10 15 30 60 and 120minutes

The foaming capacity was determined by the followingequation

FC () = VF3010158401015840

VF0times 100 (6)

where VF3010158401015840 is the volume of foam measured after 30seconds of rest after the sample has been shaken VF0 is thevolume of foam measured after agitation

The FS was determined using the following equation

FS () = VF11990511990910158401015840

VF0times 100 (7)

where VF11990511990910158401015840 is the foam volume after standing for 5 10 1530 60 and 120min

236 Gelling Minimal Concentration (GMC) The GMC ofthe protein isolates was determined according to the methoddescribed by Cheung and Chau [22] Suspensions of theprotein isolates (50mL) were prepared at a concentration of4ndash20 (wv) in distilled water and the pH was adjusted to 7by addingHCl (01 N) orNaOH (01 N)The suspensionswerepoured in 50mL falcon tubes which were placed in a vesselwith water at 100∘C for 1 h Immediately thereafter they weremaintained at 4∘C for 2 h After the cooling time the tubeswere inverted to determine the degree of displacement of thesuspensions through the walls of the tube The absence ofdisplacement at the different concentrations was consideredas the GMC in the isolates

24 Statistical Analysis The results obtained were subjectedto analyses of variance (ANOVA) using the StatgraphicsPlus software (v41) Tukeyrsquos test was used to determine thesignificance of differences between treatments with 119875 lt 005taken to indicate a significant difference

3 Results and Discussion

31 Chemical Composition of Lupine Seed Flour and ProteinConcentration of Isolates Table 1 shows the results of thechemical analysis of the different flours of lupine seeds(Haags Blaue Sonate Probor Borlu Boregine and Boruta)The results showed that the main components of the seeds


are carbohydrates and proteins (2840ndash3661) of whichthe high content is similar to that of soybean seeds [23]Significant differences (Table 1) were found between Proborand the other varieties which showed the highest proteincontent (3661) In relation to the ash content no significantdifferences (119875 gt 005) were found These values were lowerthan those reported for L luteus and similar to that of Langustifolius [24] The results of the crude fat determinationshowed lower values than those reported for soybean [25]The varieties Haags Blaue (440) Boregine (388) andBoruta (531) showed significant difference (119875 lt 005)among themselves The results show that the fiber contentvaried from 805 to 1512 in the flours of lupine seeds whichare similar to those of other legumes such as peas beans andlentils (136ndash289) [26ndash28] Regarding the carbohydratescontent the values found ranged from 4224 to 5168similar to those reported to peas beans and lentils [26] Themoisture values of the varieties of the present work wereinferior to other legumes such as common bean (10) [28]and higher to the Pardina lentil (4) [29]

The final concentrations of proteins of Haags BlaueBoregine Borlu Probor Sonate and Boruta obtained were8933 8923 8942 9061 8950 and 8840 respec-tively It has been observed that both the purity and theamount of protein recovered are affected by the characteris-tics of the seeds extraction time and temperature and therelation between the flour with the solvent and the pH atwhich protein precipitation takes place [30]


321 Solubility of Protein The solubility of protein isolatesprovides important information about their possible tech-nological applications [31] According to Horax et al [32]the solubility of the protein at different pH values can serveas a useful indicator of the efficiency or performance ofprotein isolates in food and also of the degree of proteindenaturation due to heat or chemical treatment Figure 1shows the variations of the solubility percentage of the proteinin a range frompH20 to 100 in which a similar behavior canbe observed in all isolatesThe results at pH 4 and 5 showed asharp decrease in the solubility for all six isolates of Lupinus asreportedChau et al [33] At pH 2 about 98-99 of the proteinisolates were soluble and about 98-99were soluble at pH of9 and 10 respectively with no significant differences betweenthem meanwhile the minimum solubility percentages (1-2) were recorded at pH of 40 and 50 as the isoelectricpoint of proteins At this pH the electrostatic intermolecularrepulsion and its ionic hydration at this point are minimalwhich causes precipitation of the proteins The solubilitypattern recorded in this study is similar to that reportedin isolates obtained from L albus and L spp where theminimum solubility value of the protein was less than 1at pH 40 [34] However El-Adawy et al [35] reported adifferent behavior in Lupinus termis and Lupinus albus sincethe minimum solubility value of the proteins (14 to 18) wasfound in a range of pH of 40ndash50 On the other hand Lqariet al [36] evaluated the solubility of the protein expressedas a percentage of soluble nitrogen at pH values 38ndash50 in a

ab

abbccd

ab

abc

ab

ab ab ab

abc

bc

ab ab abab

Haags BlaueBoregineBorlu

Probor

ns

SonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

100

Solu

bilit

y (

)

a

a

b

b bb

aacd

bbaa

a a acb

a

a

c

d e

b

ab ab bc bcbc

abab

Figure 1 Effect of pH on the solubility of six varieties of Lupinusangustifolius protein isolates Different letters indicate significantdifferences among varieties at119875 lt 005 ns indicates nonsignificance(119875 gt 005)

L angustifolius isolate the solubility of the protein was min-imal (225) at pH 43 In other legumes such as pea (Pisumsativum) bean (Vicia faba) and soybean (Glycine max) thelowest solubility percentage of the protein (1ndash10) was foundin pH values of 40ndash50 [37] In a similar investigation ofthe present study but with different varieties of chickpeaKaur and Singh [38] and Withana-Gamage et al [39] foundthat protein solubility was minimal (2ndash6) in a pH rangeof 40ndash45 The results showed good solubility in both theacidic and alkaline pH regions which was an importantcharacteristic in food formulations

322Water-HoldingCapacity (WHC) andOilHoldingCapac-ity (OHC) Table 2 shows the WHC and OHC of proteinisolates of Lupinus Themaximum values of water absorptionwere recorded in the isolates obtained from the Haags Blaueand Boruta varieties with 298 and 295mL gminus1 respectivelyHaags Blaue Probor Sonate and Boruta did not showsignificant differences (119875 gt 005) In general theWHCvaluesobtained in this study are considered high and are higher thanthose recorded in protein isolates of L campestris L albusL splendens L termis and L angustifolius [34 36 40] andsimilar (Mean of 246mL gminus1) to those found in commercialsoy protein isolates (Supro 670)This property is related to theability of proteins to hydrate [18] and it is important in foodsystems due to their effects on the taste and texture of food[41] The water retention is a function of isolate obtentionmethod where alkaline conditions have an influence in theprotein denaturation its solubility and the lower WHC [42]

The OHC did not show significant differences (119875 gt 005)between the isolates (Table 2) The mean value of OHC inthe six isolates was 263mL gminus1 which is higher than that


Table 2 Functional properties of protein isolates obtained from seeds of varieties of Lupinus angustifolius cultivated in Mexico

Variety Haags Blaue Boregine Borlu Probor Sonate BorutaSolubility () 031a 033a 031a 034a 031a 032a

WHC (mL gminus1) 298a 284b 281b 291a 293a 295a

OHC (mL gminus1) 265a 264a 261a 262a 263a 265a

EAI (m2 gminus1) 271b 293a 283a 275b 264b 272b

ESI (min) 147a 124b 119c 149a 131b 129b

FC () 1163a 1166a 1165a 1167a 1164a 1168a

FS () 928ab 934a 919c 926b 924b 939a

GMC () 1100b 1000c 1100b 1200a 1100b 1100b

Different letters in the same row indicate significant differences (119875 lt 005) WHC water holding capacity OHC oil holding capacity EAI emulsion activityindex ESI emulsion stability index FC foaming capacity FS foam stability index GMC gelling minimum concentration

recorded in an isolate of L campestris (17mL gminus1) but lowerthan that obtained in isolates of L albus and L splendenswith285 and 463mL gminus1 respectively [34 40] Lqari et al [36]in other leguminous isolates such as peas (Pisum sativum)bean (Vicia faba) and soybean (Glycine max) also reportedOHC values lower (12 16 and 11mL gminus1 protein) than thoserecorded in the isolates obtained and characterized in thisstudy

In general as reported by Carvalho et al [42] the OHCobtained in this study shows that Lupinus protein isolatespossess well balanced proportions of externally orientedhydrophilic and hydrophobic groups being able to reducethe interfacial tension in an emulsion system they are veryimportant in such as applications as in processing meat basesproducts The values of WHC and OHC for these proteinisolates that can be considered high in relation to othersprotein isolates used as ingredients in the preparation of coldmeats especially for sausages are determining properties todevelop a food of acceptable quality [18]

323 Emulsion Activity Index (EAI) and Emulsion StabilityIndex (ESI) Significant differences (119875 lt 005) were observedin both indexes according to the L angustifolius varieties used(Table 2) The isolates obtained from the Boregine and Borluvarieties showed the highest EAI with 293 and 283m2 gminus1respectively while the lowest was recorded in the isolateobtained from the Sonate variety (264m2 gminus1) while in theESI the highest values were 149 and 147min in the Proborand Haags Blaue isolates respectively and while the lowerindex was found in the protein isolate obtained from theBorlu variety (119min) (119875 lt 005)

The profiles of emulsifying activities (EAI and ESI)against pH are shown in Figures 2 and 3 The profiles ofEAI and ESI are similar to solubility these properties alsoincreased at both pH extremes evaluated and were minimalin the isoelectric pH range (40 to 50) The profile of theemulsion properties as a function of pH for the differentprotein isolates studied was similar to that presented bythe Indian and soybean protein isolates [18] as well as Lcampestris isolates [40]The slight differences of the emulsionproperties in the studied varieties can be due to the differentlevel of solubilized proteins which can influence the EAI andESI by modifying the balance of Van der Waals attractive

ns

ns

ns


ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

ab ab ab

ab ab abbc

ab a

a

c

c

cab bca c

abb

cc d d aaa

b b b bb

b bb

b

EAI (

Ｇ2Ａminus1)

Figure 2 Effect of pH on the emulsion activity index (EAI) of sixvarieties of Lupinus angustifolius protein isolates Different lettersindicate significant differences among varieties at 119875 lt 005 nsindicates nonsignificance (119875 gt 005)

and repulsive electrostatic forces a more soluble proteinproduces a rapid migration to the oil-water interface thusfavoring the formation and stability of emulsions [43] Ingeneral the profiles of EAI (Figure 2) and ESI (Figure 3)presented variations similar to those reported in other isolatesor concentrates which are defined as a V shape patternindicating changes of the hydrophilic-lipophilic balance ofthe proteins in the pH gradient from 2 to 10 The resultconfirms the relationship between emulsifying and solubilityproperties of proteins However the results shown in thisstudy and similar ones should be validated with other factorsthat may influence the emulsification properties amongthem droplet size net charge interfacial tension viscosityand protein conformation [42 44] Temperature is also a keyfactor affecting emulsifying properties of proteins Duringthe present study the conditions used were 25∘C while other



ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

abab

abab

ab

ab

ab bc

bc bc

decd

c cc

c

c

e

bb

b

bbbbb

bbbb

ab abab

ab bc cdd b

b

b

a a

a

a

a a a aa a

a

a

a

a

ESI (

min

)

Figure 3 Effect of pH on the emulsion stability index (ESI) of sixvarieties of Lupinus angustifolius protein isolates Different lettersindicate significant differences among varieties at 119875 lt 005

methodologies for the determination of emulsification report80∘C [45]

324 Foaming Capacity (FC) and Foam Stability (FS) Theprotein isolates showed similar FC values between 1163 and1168 (Table 2) without significant differences (119875 gt 005)These results are similar to soybean (116) [22] but higherthan those reported in other beans [29 43 46 47] The highvalues of this property can be attributed to flexible proteinmolecules which reduce surface tension and diffuse morequickly at the air-water interface encapsulating air particlesand thus favoring the formation of foam [48] With respectto FS significant differences (119875 lt 005) were recorded amongthe six isolates evaluated (Table 2) with the highest value inthe Boruta variety (939) and lowest value in Borlu variety(919) The results for FS were similar to those reportedfor L angustifolius protein isolates [36] and higher thanthose reported for soy protein isolates [39] When the foamstability values do not differ greatly from each other it maybe due to the ability to form a cohesive viscoelastic film viaintermolecular interactions [49] According to the study ofAdebowale et al [49] the better stability of the foams mightbe attributed to formation of stable molecular layers in theacidic pH range However within food systems foams arecomplex systems including different phases such as amixtureof gases subdivided at solids liquids and multicomponentsolutions of water polymers and surfactants which makenecessary studies in a wide range of pH values [50]

325 Gelling Minimum Concentration (GMC) The proteinsare efficient gelling agents A gel can be defined as anintermediate state between solid and liquid Several factorsinfluence gel formation such as the protein size molecular

weight amino acid composition pH and interaction withother components since large molecules form extensivenetworks by cross-linking in three dimensions and also by theflexibility and ability of the proteins to undergo denaturationThe results obtained forGMCshow that the protein isolates ofthe different lupine varieties need a minimum concentrationof protein that ranged from 10 (Boregine) up to 12(Probor) in order to form a stable gel (Table 2) The resultsare similar to those found in the literature for L angustifoliusprotein isolates [36] in common varieties [29 47] The GMCof Boregine was slightly better than soybean protein isolate(10) However it is observed that lentil [29] and lima beans[47] show higher gelling capacity (7) while other types ofbeans [43] have a less gelling capacity (15)These variationsbetween the different legumes seem to be related to thecontent of globulins which have high ease of forming gels[50 51] Hence protein isolates of Lupinus are particularlyuseful in preparing comminuted meat emulsion as reportedby Uvarova and Barrera-Arellano [44]

4 Conclusions

The chemical composition of L angustifolius flour showed asignificant difference among the different varieties evaluatedwith protein contents of 2840 in Boruta and 3661 inProbor Functional properties such as nitrogen solubility andthe indexes of emulsion activity and emulsion stability wereaffected by the pH of the medium In general all these wereminimal at values near the isoelectric point of the proteins(pH 4ndash6) and reached their maximum values at acidic (2)and alkaline pH (10) The maximum solubility of the proteinisolates (99) was observed at pH values of 20 and 100 with-out significant differences between varieties Water-holdingcapacity showed significant differences between varietieswhile oil holding capacity and foam formation were similarin all varieties

Conflicts of Interest

The authors declare that there are no conflicts of interestregarding the publication of this paper

Acknowledgments

The authors acknowledge the National Council of Scienceand Technology of Mexico that granted Scholarship 589981to Antonio Hilario Lara-Rivera

References

[1] H Schumacher H M Paulsen A E Gau et al ldquoSeed proteinamino acid composition of important local grain legumes Lupi-nus angustifolius L Lupinus luteus L Pisum sativum L andVicia faba Lrdquo Plant Breeding vol 130 no 2 pp 156ndash164 2011

[2] M Saastamoinen M Eurola and V Hietaniemi ldquoThe chemicalquality of some legumes peas fava beans blue and white lupinsand soybeans cultivated in Finlandrdquo Journal of AgriculturalScience and Technology vol 3 no 3 pp 92ndash100 2013

[3] L Lopez-Bellido and M Fuente ldquoLupin crop as an alternativesource of proteinrdquo Advances in Agronomy vol 40 pp 239ndash2951986


[4] C I Lizarazo A-M Lampi J Liu T Sontag-Strohm VPiironen and F L Stoddard ldquoNutritive quality and protein pro-duction from grain legumes in a boreal climaterdquo Journal of theScience of Food and Agriculture vol 95 no 10 pp 2053ndash20642015

[5] G D Hill ldquoThe comparison of nutritive value of lupin seedrdquoNutrition Abstracts and Reviews vol 47 pp 511ndash529 1977

[6] D S Petterson ldquoThe use of lupins in feeding systemsmdashreviewrdquoAsian-Australasian Journal of Animal Sciences vol 13 no 6 pp861ndash882 2000

[7] A Sujak A Kotlarz andW Strobel ldquoCompositional and nutri-tional evaluation of several lupin seedsrdquo Food Chemistry vol98 no 4 pp 711ndash719 2006

[8] L Yeheyis C Kijora E Van Santen M Wink and K J PetersldquoSweet annual lupins (Lupinus spp) their adaptability and pro-ductivity in different agro-ecological zones of Ethiopiardquo Journalof Animal Science Advances vol 2 no 2 pp 201ndash215 2012

[9] G G Fontanari J M Martins M Kobelnik et al ldquoThermalstudies on protein isolates of white lupin seeds (Lupinus albus)rdquoJournal of Thermal Analysis and Calorimetry vol 108 no 1 pp141ndash148 2012

[10] M Duranti A Consonni C Magni F Sessa and A ScarafonildquoThemajor proteins of lupin seed Characterisation and molec-ular properties for use as functional and nutraceutical ingredi-entsrdquo Trends in Food Science amp Technology vol 19 no 12 pp624ndash633 2008

[11] B Vargas-Guerrero P M Garcıa-Lopez A L Martınez-AyalaJ A Domınguez-Rosales and C M Gurrola-Dıaz ldquoAdmin-istration of Lupinus albus Gamma Conglutin (C120574) to n5 STZRats Augmented Ins-1 Gene Expression and Pancreatic InsulinContentrdquo Plant Foods for Human Nutrition vol 69 no 3 pp241ndash247 2014

[12] C Patane E Iacoponi and S A Raccuia ldquoPhysico-chemicalcharacteristics water absorption soaking and cooking proper-ties of some Sicilian populations of chickpea (Cicer arietinumL)rdquo International Journal of Food Sciences and Nutrition vol55 no 7 pp 547ndash554 2004

[13] N G Vera ldquoUtilizacion de los derivados de cereales y legumi-nosas en la elaboracion de productos carnicosrdquoNacameh vol 1no 1 pp 110ndash117 2007

[14] E GarcıaModificaciones al sistema de clasificacion climatica deKoppen 1998

[15] AOAC Official Methods of Analysis Association of OfficialAnalytical Chemists Washington DC USA 2000

[16] J Vioque V R Sanchez J Pedroche Y M del Mar and FMillan ldquoObtencion y aplicaciones de concentrados y aisladosproteicosrdquo Grasas y Aceites vol 52 no 2 pp 127ndash131 2001

[17] C Martınez-Villaluenga G Urbano J M Porres J Frias andC Vidal-Valverde ldquoImprovement in food intake and nutritiveutilization of protein from Lupinus albus var multolupa proteinisolates supplemented with ascorbic acidrdquo Food Chemistry vol103 no 3 pp 944ndash951 2007

[18] S O Ogunwolu F O Henshaw H-P Mock A Santros andS O Awonorin ldquoFunctional properties of protein concentratesand isolates produced from cashew (Anacardium occidentale L)nutrdquo Food Chemistry vol 115 no 3 pp 852ndash858 2009

[19] X Liu L Zhang S Wei S Chen X Ou and Q Lu ldquoOveroxi-dized polyimidazolegraphene oxide copolymer modified elec-trode for the simultaneous determination of ascorbic aciddopamine uric acid guanine and adeninerdquo Biosensors andBioelectronics vol 57 pp 232ndash238 2014

[20] C-F Chau Y-L Huang andM-H Lee ldquoIn vitro hypoglycemiceffects of different insoluble fiber-rich fractions prepared fromthe peel ofCitrus sinensis L cv liuchengrdquo Journal of Agriculturaland Food Chemistry vol 51 no 22 pp 6623ndash6626 2003

[21] H-G Zheng X-Q Yang C-H Tang L Li and I AhmadldquoPreparation of soluble soybean protein aggregates (SSPA) frominsoluble soybean protein concentrates (SPC) and its functionalpropertiesrdquo Food Research International vol 41 no 2 pp 154ndash164 2008

[22] P C-K Cheung and C-F Chau ldquoChanges in the dietary fiber(resistant starch and nonstarch polysaccharides) content ofcooked flours prepared from three Chinese indigenous legumeseedsrdquo Journal of Agricultural and Food Chemistry vol 46 no 1pp 262ndash265 1998

[23] M Bahr A Fechner K Hasenkopf S Mittermaier and GJahreis ldquoChemical composition of dehulled seeds of selectedlupin cultivars in comparison to pea and soya beanrdquo LWT- FoodScience and Technology vol 59 no 1 pp 587ndash590 2014

[24] A de Luna-Jimenez ldquoComposicion y procesamiento de la soyapara consumo humanordquo Investigacion y Ciencia de la Univer-sidad Autonoma de Aguascalientes vol 15 no 37 pp 35ndash442007

[25] G E De Almeida Costa K Da Silva Queiroz-Monici S M PM Reis and A C De Oliveira ldquoChemical composition dietaryfibre and resistant starch contents of raw and cooked peacommon bean chickpea and lentil legumesrdquo Food Chemistryvol 94 no 3 pp 327ndash330 2006

[26] Z-U Rehinan M Rashid and W H Shah ldquoInsoluble dietaryfibre components of food legumes as affected by soaking andcooking processesrdquo Food Chemistry vol 85 no 2 pp 245ndash2492004

[27] K R Sridhar and S Seena ldquoNutritional and antinutritionalsignificance of four unconventional legumes of the genusCanavalia - A comparative studyrdquo Food Chemistry vol 99 no2 pp 267ndash288 2006

[28] E Marconi S Ruggeri M Cappelloni D Leonardi and ECarnovale ldquoPhysicochemical nutritional and microstructuralcharacteristic of chickpeas (Cicer arietinum L) and commonbeans (Phaseolus vulgaris L) following microwave cookingrdquoJournal of Agricultural and Food Chemistry vol 48 no 12 pp5986ndash5994 2000

[29] G Y Aguilera Harinas Leguminosas deshidratadas Carac-terizacion Nutricional y valoracion de sus propiedades tecno-funcionales [Tesis Doctoral] Universidad Autonoma deMadridFacultad de Ciencias Departamento de Quımica Agrıcola2009

[30] J Boye F Zare and A Pletch ldquoPulse proteins processingcharacterization functional properties and applications in foodand feedrdquo Food Research International vol 43 no 2 pp 414ndash431 2010

[31] E Lampart-Szczapa P Konieczny M Nogala-Kałucka S Wal-czak I Kossowska and M Malinowska ldquoSome functionalproperties of lupin proteinsmodified by lactic fermentation andextrusionrdquo Food Chemistry vol 96 no 2 pp 290ndash296 2006

[32] R Horax N S Hettiarachchy P Chen and M JalaluddinldquoPreparation and characterization of protein isolate from cow-pea (Vigna unguiculata L Walp)rdquo Journal of Food Science vol69 no 2 pp FCT114ndashFCT118 2004

[33] C-F Chau P C K Cheung and Y-S Wong ldquoFunctional prop-erties of protein concentrates from three Chinese indigenouslegume seedsrdquo Journal of Agricultural and Food Chemistry vol45 no 7 pp 2500ndash2503 1997


[34] J Porras-Saavedra N Guemez-Vera J L Montanez-Soto andM Carmen ldquoComparative study of functional properties ofprotein isolates obtained from three Lupinus speciesrdquo Advancesin Bioresearch vol 4 pp 106ndash116 2013

[35] T A El-Adawy E H Rahma A A El-Bedawey and A F GafarldquoNutritional potential and functional properties of sweet andbitter lupin seed protein isolatesrdquo Food Chemistry vol 74 no 4pp 455ndash462 2001

[36] H Lqari J Vioque J Pedroche and F Millan ldquoLupinus angus-tifolius protein isolates Chemical composition functionalproperties and protein characterizationrdquo Food Chemistry vol76 no 3 pp 349ndash356 2002

[37] A Fernandez-Quintela M TMacarulla A S Del Barrio and JAMartınez ldquoComposition and functional properties of proteinisolates obtained from commercial legumes grown in northernSpainrdquo Plant Foods for Human Nutrition vol 51 no 4 pp 331ndash342 1997

[38] M Kaur and N Singh ldquoCharacterization of protein isolatesfrom different Indian chickpea (Cicer arietinum L) cultivarsrdquoFood Chemistry vol 102 no 1 pp 366ndash374 2007

[39] T S Withana-Gamage J P Wanasundara Z Pietrasik and PJ Shand ldquoPhysicochemical thermal and functional characteri-sation of protein isolates from Kabuli and Desi chickpea (Cicerarietinum L) A comparative study with soy (Glycine max) andpea (Pisum sativum L)rdquo Journal of the Science of Food andAgriculture vol 91 no 6 pp 1022ndash1031 2011

[40] S L Rodrıguez-Ambriz A L Martınez-Ayala F Millan andG Davila-Ortız ldquoComposition and functional properties ofLupinus campestris protein isolatesrdquo Plant Foods for HumanNutrition vol 60 no 3 pp 99ndash107 2005

[41] J Yu M Ahmedna and I Goktepe ldquoPeanut protein con-centrate Production and functional properties as affected byprocessingrdquo Food Chemistry vol 103 no 1 pp 121ndash129 2007

[42] A V Carvalho N H P Garcıa and J Amaya-Farfan ldquoPhysico-chemical properties of the flour protein concentrate and pro-tein isolate of the cupuassu (Theobroma grandiflorum Schum)seedrdquo Journal of Food Science vol 71 no 8 pp S573ndashS578 2006

[43] K O Adebowale and O S Lawal ldquoComparative study of thefunctional properties of bambarra groundnut (Voandzeia sub-terranean) jack bean (Canavalia ensiformis) and mucuna bean(Mucuna pruriens) floursrdquo Food Research International vol 37no 4 pp 355ndash365 2004

[44] M L Uvarova and D Barrera-Arellano ldquoEfecto de la lipofil-izacion sobre las propiedades funcionales de la harina depalmist (Elaeis guineensis)rdquo Grasas y Aceites vol 56 no 1 pp1ndash8 2005

[45] F H Brishti S K S Zarei M R Ismail-Fitry R Shukri andN Saari ldquoEvaluation of the functional properties of mung beanprotein isolate for development of textured vegetable proteinrdquoInternational Food Research Journal vol 24 no 4 pp 1595ndash1605 2017

[46] L Chel-Guerrero V Perez-Flores D Betancur-Ancona and GDavila-Ortiz ldquoFunctional properties of flours and protein iso-lates from Phaseolus lunatus and Canavalia ensiformis seedsrdquoJournal of Agricultural and Food Chemistry vol 50 no 3 pp584ndash591 2002

[47] E Sangronis ldquoPropiedades funcionales de las harinas deleguminosas (Phaseolus vulgaris y Cajan cajan) germinadasrdquoInterciencia vol 29 no 2 pp 80ndash85 2004

[48] R E Aluko and R Y Yada ldquoStructure-function relationshipsof cowpea (Vigna unguiculata) globulin isolate influence of pH

and NaCl on physicochemical and functional propertiesrdquo FoodChemistry vol 53 no 3 pp 259ndash265 1995

[49] Y A Adebowale I A Adeyemi and A A Oshodi ldquoFunc-tional and physicochemical properties of flours of six MucunaspeciesrdquoAfrican Journal of Biotechnology vol 4 no 12 pp 1461ndash1468 2005

[50] S K Sathe and D K Salunkhe ldquoFunctional properties of thegreat northern bean (Phaseolus vulgaris L) proteins emulsionfoaming viscosity and gelation propertiesrdquo Journal of FoodScience vol 46 no 1 pp 71ndash81 1981

[51] S K Sathe S S Deshpande and D K Salunkhe ldquoFunctionalproperties of lupin seed (Lupinus mutabilis) proteins andprotein concentratesrdquo Journal of Food Science vol 47 no 2 pp491ndash497 1982

Submit your manuscripts athttpswwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 201


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


information on how a particular ingredient would behave ina food matrix play a prominent role during the preparationprocessing and storage of food and in turn correlate withsensory properties These properties could be considered asa result of different conformational changes or interactionsbetween food components such as interactions betweenproteins proteins and polysaccharides lipids phenolic com-pounds or phytic acid [13] Furthermore they constitute thefunctional base of diverse products mainly of low proteincontent and high fat content On the other hand legumesare source of beneficial compounds that have a protectiveeffect on the development of various diseases [14] Thereforethe objective of the present research work was to determinethe functional properties of protein isolates of six varietiesobtained from Lupinus angustifolius cultivated in the state ofJalisco Mexico

2 Material and Methods

21 Vegetable Materials Seeds of six varieties of L angus-tifolius (Haags Blaue Sonate Probor Borlu Boregine andBoruta) were used which were provided by the Germancompany Saatzucht Steinach GmbH dedicated to geneticimprovement These seeds were sown and cultivated duringthe fallwinter period of 20142015 under irrigated conditionson agricultural soils of the University Center of Biologicaland Agricultural Sciences of the University of Guadalajara inZapopan Jalisco MexicoThis site is located at the geograph-ical coordinates 20∘4410158404710158401015840N and 103∘3010158404310158401015840W at an altitudeof 1523m It is characterized by a humid temperate climatewith rains in summer with precipitations ranging from 700to 1400mm per year and an annual average temperature of120 to 180∘C with the presence of frost After conventionalland preparation with a moldboard plow and a disk harrowseeding was performed manually under an experimentalrandomized block design with four replicates

22 Proximate Composition and Preparation of Protein Iso-lates After harvesting on the order of 50 kg of seeds onebatch of 1 kg each variety was separately ground with a knifemill (IKA) until flour with a particle size of 2mm wasobtained Every variety was subjected to triplicate a proximalchemical analysis according to the techniques described inAOAC (2000) moisture content (92509) crude fat (92030)crude protein (97909) ash (92303) crude fiber (96209)and carbohydrate content [15] The flour was subjected toa degreasing process by Soxhlet extraction using hexane assolvent [16] The defatted sample (100 g) was used to obtainthe protein isolates

The protein isolate was obtained using the isoelectricprecipitation method [17] A dispersion of 100 g Lminus1 inNa2SO3(0025) was prepared and adjusted to pH of 90

using NaOH (1N) Immediately it was stirred for one hourat room temperature (25∘C) and centrifuged at 10976timesg(HERMLELabortechnikGmbHZ 326Germany) for 10minThe supernatant was acidified to pH of 45 using HCl (1 N)This was subjected to centrifugation at 10976timesg for 10minat 4∘C recovering the precipitate A washing process withdistilled water was applied to the precipitate and NaOH

(1N) was used to bring it to a pH of 7 The precipitate waslyophilized and stored at 4∘C The protein isolates obtainedfrom the seeds of the six varieties were analyzed in triplicatefor all functional properties


231 Solubility Thesolubility of the protein isolates was eval-uated according to the methodology proposed by Ogunwoluet al [18] and Liu et al [19] with slight modifications Eachisolate was prepared in a 01 wv suspension in 10mL ofdistilled water The pH was adjusted in a range of 20ndash100using NaOH (01 N) or HCl (01 N) The suspension wasstirred for 30min on an orbital shaker (Thermo Fisher Sci-entific Multipurpose rotator 2346 China) Subsequently itwas centrifuged for 10min at 10976timesg (HERMLELabortech-nik GmbH Z 326 Germany) The protein content in thesupernatant was determined by the Bradford method Thestandard curve was made from bovine serum albumin (BSA)and the absorbance of the samples was measured at 595 nmon a spectrophotometer (Thermo Fisher Scientific G10S Uv-Vis China)The solubility of the protein was calculated by thefollowing equation

Solubility =protein in supernatant (g)protein in sample (g)

times 100 (1)

232 Water-Holding Capacity (WHC) The WHC was eval-uated by the method of Chau et al [20] with slight modi-fications Samples of protein isolates (05 g) were dispersedin 5mL of distilled water and stirred for 24 h After thistime the centrifugation for 30min at 1474 g (HERMLELabortechnik GmbH Z 326 Germany) was carried out Thevolume of the supernatant wasmeasured after centrifugationThe difference between the initial volume of water and thevolume recovered after centrifugation determines the WHCexpressed as milliliters of water by one gram of sample(mL gminus1)

TheWHC was determined using the following equation

WHC (mLgminus1) = 119881119894 minus 119881119904119882119898

(2)

where119881119894is the initial volume of distilled water (mL)119881

119904is the

volume of the supernatant (mL) and119882119898is the weight of the

sample (g)

233 Oil Holding Capacity (OHC) The OHC was evaluatedby the method of Chau et al [20] with slight modifications05 g of the protein isolates that were obtained was added5mL of canola oil contained in a 50ml centrifuge tubewhich was stirred for 30min by magnetic stirring Then it iscentrifuged at 1474 g (HERMLE Labortechnik GmbH Z 326Germany) for 30 minutes After centrifugation the volumeof the supernatant is measured The difference between theinitial volume of oil and the volume recovered correspondsto the OHC expressed in mL gminus1 of protein isolates




Boregine 365b 2923c 334a 388c 822c 5168a

Borlu 438a 3342b 313a 417b 1006b 4484b

Probor 384b 3661a 325a 420b 986ab 4224b

Sonate 379b 3250b 351a 529a 1249a 4242a

Boruta 368b 2840c 343a 531a 1512a 4406a




(3)







(4)



10) The


ESI (min) =1198600

1198600minus 11986010

times 10 (5)




FC () = VF3010158401015840

VF0times 100 (6)



FS () = VF11990511990910158401015840

VF0times 100 (7)











ab

abbccd

ab

abc

ab

ab ab ab

abc

bc

ab ab abab


Probor

ns

SonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

100

Solu

bilit

y (

)

a

a

b

b bb

aacd

bbaa

a a acb

a

a

c

d e

b

ab ab bc bcbc

abab











ESI (min) 147a 124b 119c 149a 131b 129b

FC () 1163a 1166a 1165a 1167a 1164a 1168a

FS () 928ab 934a 919c 926b 924b 939a

GMC () 1100b 1000c 1100b 1200a 1100b 1100b






ns

ns

ns


ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

ab ab ab

ab ab abbc

ab a

a

c

c

cab bca c

abb

cc d d aaa

b b b bb

b bb

b

EAI (

Ｇ2Ａminus1)





ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

abab

abab

ab

ab

ab bc

bc bc

decd

c cc

c

c

e

bb

b

bbbbb

bbbb

ab abab

ab bc cdd b

b

b

a a

a

a

a a a aa a

a

a

a

a

ESI (

min

)






4 Conclusions




Acknowledgments


References






























































Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




Boregine 365b 2923c 334a 388c 822c 5168a

Borlu 438a 3342b 313a 417b 1006b 4484b

Probor 384b 3661a 325a 420b 986ab 4224b

Sonate 379b 3250b 351a 529a 1249a 4242a

Boruta 368b 2840c 343a 531a 1512a 4406a




(3)







(4)



10) The


ESI (min) =1198600

1198600minus 11986010

times 10 (5)




FC () = VF3010158401015840

VF0times 100 (6)



FS () = VF11990511990910158401015840

VF0times 100 (7)











ab

abbccd

ab

abc

ab

ab ab ab

abc

bc

ab ab abab


Probor

ns

SonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

100

Solu

bilit

y (

)

a

a

b

b bb

aacd

bbaa

a a acb

a

a

c

d e

b

ab ab bc bcbc

abab











ESI (min) 147a 124b 119c 149a 131b 129b

FC () 1163a 1166a 1165a 1167a 1164a 1168a

FS () 928ab 934a 919c 926b 924b 939a

GMC () 1100b 1000c 1100b 1200a 1100b 1100b






ns

ns

ns


ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

ab ab ab

ab ab abbc

ab a

a

c

c

cab bca c

abb

cc d d aaa

b b b bb

b bb

b

EAI (

Ｇ2Ａminus1)





ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

abab

abab

ab

ab

ab bc

bc bc

decd

c cc

c

c

e

bb

b

bbbbb

bbbb

ab abab

ab bc cdd b

b

b

a a

a

a

a a a aa a

a

a

a

a

ESI (

min

)






4 Conclusions




Acknowledgments


References






























































Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology






ab

abbccd

ab

abc

ab

ab ab ab

abc

bc

ab ab abab


Probor

ns

SonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

100

Solu

bilit

y (

)

a

a

b

b bb

aacd

bbaa

a a acb

a

a

c

d e

b

ab ab bc bcbc

abab











ESI (min) 147a 124b 119c 149a 131b 129b

FC () 1163a 1166a 1165a 1167a 1164a 1168a

FS () 928ab 934a 919c 926b 924b 939a

GMC () 1100b 1000c 1100b 1200a 1100b 1100b






ns

ns

ns


ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

ab ab ab

ab ab abbc

ab a

a

c

c

cab bca c

abb

cc d d aaa

b b b bb

b bb

b

EAI (

Ｇ2Ａminus1)





ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

abab

abab

ab

ab

ab bc

bc bc

decd

c cc

c

c

e

bb

b

bbbbb

bbbb

ab abab

ab bc cdd b

b

b

a a

a

a

a a a aa a

a

a

a

a

ESI (

min

)






4 Conclusions




Acknowledgments


References






























































Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology







ESI (min) 147a 124b 119c 149a 131b 129b

FC () 1163a 1166a 1165a 1167a 1164a 1168a

FS () 928ab 934a 919c 926b 924b 939a

GMC () 1100b 1000c 1100b 1200a 1100b 1100b






ns

ns

ns


ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

90

ab ab ab

ab ab abbc

ab a

a

c

c

cab bca c

abb

cc d d aaa

b b b bb

b bb

b

EAI (

Ｇ2Ａminus1)





ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

abab

abab

ab

ab

ab bc

bc bc

decd

c cc

c

c

e

bb

b

bbbbb

bbbb

ab abab

ab bc cdd b

b

b

a a

a

a

a a a aa a

a

a

a

a

ESI (

min

)






4 Conclusions




Acknowledgments


References






























































Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



ProborSonateBoruta

113 4 5 6 7 8 9 1021

pH

0

10

20

30

40

50

60

70

80

abab

abab

ab

ab

ab bc

bc bc

decd

c cc

c

c

e

bb

b

bbbbb

bbbb

ab abab

ab bc cdd b

b

b

a a

a

a

a a a aa a

a

a

a

a

ESI (

min

)






4 Conclusions




Acknowledgments


References






























































Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



























































Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




























Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 201






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

Functional Properties of Lupinus angustifolius Seed …downloads.hindawi.com/journals/jfq/2017/8675814.pdfSeed Protein Isolates AntonioHilarioLara-Rivera, 1 PedroGarcía-Alamilla,