采后杏果中果胶酶和螯合可溶果胶的交互作用的体外研究.pdf
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1、Original articleExtraction and physicochemical properties of soya bean proteinand oil by a new reverse micelle system compared with otherextraction methodsGuanhao Bu,1Yingying Yang,1Fusheng Chen,1*Zhixiong Liao,2Yanxiu Gao,1Hongshun Yang,3,4Runjie Li,1Kunlun Liu1&Junting Zhao51 College of Food Scien
2、ce and Technology,Henan University of Technology,Zhengzhou 450001,China2 Xinke College,Henan Institute of Science and Technology,Xinxiang 453003,China3 Food Science and Technology Programme,Department of Chemistry,National University of Singapore,Science Drive 2,Singapore 117543,Singapore4 National
3、University of Singapore(Suzhou)Research Institute,Suzhou Industrial Park,377 Lin Quan Street,Jiangsu 215123,China5 School of Chemistry and Chemical Engineering,Henan University of Technology,Zhengzhou 450001,China(Received 22 April 2013;Accepted in revised form 25 September 2013)SummaryReverse micel
4、le extraction is a novel technology for the separation of plant components such as proteinsand oil.In this study,sodium bis(2-ethylhexyl)sulphosuccinate(AOT)reverse micelle system and AOT/Tween 85 reverse micelle system were used to extract soya bean protein and oil from soya bean flour.Thephysicoch
5、emical properties of the protein and oil extracted were investigated and compared with tradi-tional extraction methods.The results showed that the efficiency of forward extraction of soya bean pro-tein using an AOT/Tween 85 reverse micelle system was superior to that using an AOT reverse micellesyst
6、em at the optimal extraction conditions.In addition,soya bean proteins extracted using reversemicelle extraction had no unordered structure under Fourier transform infrared spectroscopy.The acidand peroxide values of oil products from two reverse micelle extractions were lower than that fromimmersio
7、n.The results indicated that AOT/Tween 85 reverse micelle system is effective in extracting soyabean protein and oil.KeywordsAOT,reverse micelle extraction,soya bean oil,soya bean protein,Tween 85.IntroductionReverse micelle extraction,a relatively new technologyfor liquidliquid extraction,especiall
8、y for the isolationand purification of proteins,has recently received dueattention(Luisi et al.,1979;Harikrishna et al.,2002).Reverse micelles are formed by surfactants in nonpolarorganic solvents,while in their polar cores,nanometre-sized water pools are formed by the solubilisation inwater(Luisi e
9、t al.,1979;Naoea et al.,2011).Withself-assembly capacity,reverse micelle systems are notonly dynamic balancing systems but also stable andtransparent thermodynamic systems(Liu et al.,2006).The overall mechanism of reverse micelle extractionconsists of two fundamental steps:forward extraction(protein
10、 is transferred into polar water pools and oil istransferred into the organic phase)and backwardextraction(protein is released from polar water poolsand transferred into an aqueous phase to be recovered,resulting in separation of aqueous phase and oil phase)(Zhao et al.,2010a).The polar water pools
11、inside reverse micelles cansolubilise hydrophilic biomolecules,such as proteins,enzymes,DNA,nucleic acids,short peptides andamino acids.The size of water pools plays a significantrole in the solubility of biomolecules in the micellecore.W0,the determining factor of the size of waterpools and the mol
12、ar ratio,is the ratio of water andsurfactant,which can be adjusted by an aqueous buffercontaining an appropriate amount of salt(Ghazi et al.,2006).Reverse micelles form a three-phase systemincluding watersurfactantorganic solvent(i.e.theform of W/O(water/oil)with various shapes,such asspherical,oval
13、 and rod);thus in this system,the bio-molecules inside the polar water pools are protectedfrom denaturation by organic solvents.Reverse micelles are more suitable for separatingproteins than regular liquidliquid extraction or otherseparation methods because in those methods,the trans-*Correspondent:
14、Fax:+86 371 67756667;e-mail:International Journal of Food Science and Technology 2014,49,10791089doi:10.1111/ijfs.12403 2013 Institute of Food Science and Technology1079fer of proteins into solvents often results in irreversibledenaturation and loss of biological activity(Guo et al.,2008).Luisi et a
15、l.(1979)first pointed out the potentialof reverse micelle solubilisation of proteins for proteinseparation.Dekker et al.(1986)and Goklen&Hatton(1985)developed the process systematically for practicaluse of reverse micelle systems.Later on,many research-ers demonstrated the factors including molar ra
16、tio W0,aqueous phase pH and ionic strength(Rho&Kang,2004),surfactant type and concentration(Shin&Vera,2002),and co-surfactant(Lee et al.,2004)affecting pro-tein solubilisation based on the interactions betweenreverse micelles and proteins.Among these parameters,surfactant is well known as playing an
17、 important role instabilising protein solubilisation in reverse micelles.Most studies to date have focused on the extraction ofproteins by reverse micelle systems,whereas studies onthe extraction of oil have been rarely reported.With the development of protein extraction by ionicreverse micelles,pro
18、tein sectional deactivation causedby the strong electrostatic interaction occasionallyoccurs in the extraction process(Harikrishna et al.,2002).Therefore,some nonionic surfactants are usedto form nonionic reverse micelles for protein extrac-tion to avoid protein deactivation.Tween 85 is usuallyused
19、for reverse micelle extractions,and it does nothas detrimental effects on the structure,function andstability of proteins solubilised in reverse micelles(Sawada et al.,2004).However,the extraction effi-ciency by Tween 85 itself was very low.In this study,the efficiency of two reverse micellesystems,
20、ananionicsurfactantnonionicsurfactantAOT/Tween 85 mixed system and an anionic surfac-tant AOT system in extracting protein and oil fromsoya bean flour,was investigated.Effects of variousfactors including soya bean flour concentration,W0,temperature,time,pH,ionic strength and ultrasonicpower on the e
21、fficiency of forward extraction of soyabean protein were examined.The physical and chemi-cal properties of oil extracted by reverse micelle werecompared with oil immersed,such as acid value(AV),peroxide value(POV)and fat acid(FA)composition.Materials and methodsMaterialsChemically pure AOT and Tween
22、 85 were purchasedfrom Shanghai Haiqu Chemical Co.Ltd.(Shanghai,China)andShanghaiLichenLimitedCompany(Shanghai,China),respectively.All other reagentsused were of analytical grade.Full-fat soya bean floursieved through a 100-mesh screen and containing42.2%protein,4.3%moisture and 24.7%oil wasobtained
23、 from Anyang Mantianxue Food Manufactur-ing Co.Ltd(Anyang,Henan,China).Proximate composition analysisCrude protein of soya bean flour was determined usingthe micro-Kjeldahl method(Concon&Soltess,1973),whereas crude fat was determined by Soxhlet extrac-tion(AOAC,1984).The moisture content was mea-sur
24、ed by drying in an oven at 105 C until constantweight was obtained.AV,POV and FA compositionwere determined according to standard methods of theInternational Union of Pure and Applied Chemistryfor analysis of oil and fats.Preparation of two reverse micelle systemsAn AOT reverse micelle was obtained
25、by dissolvingAOT in isooctane and then injecting the mixture withKClphosphate buffer solution.A stock solution of0.08 g mL?1AOT reverse micelle system was obtainedby mixing AOT with isooctane and then stirring themixture using a magnetic stirrer at room temperature.When AOT was dissolved completely,
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