牛骨胶原蛋白酶解工艺优化及结构特性分析 .pdfVIP

牛骨胶原蛋白酶解工艺优化及结构特性分析

刘丽莉;李丹;杨陈柳;孟圆圆;代晓凝

【摘要】为研究前期纯化的特异性降解牛骨胶原蛋白的胶原蛋白酶(BSC)对牛骨胶

原蛋白的酶解工艺,以水解度为响应值,在单因素试验基础上通过五元二次正交旋转

组合试验,确定最佳酶解工艺为:反应温度46℃、牛骨胶原蛋白添加量5.14g/100

mL、BSC添加量0.42g/100mL、反应时间6h、pH6.5,该工艺条件下水解度为

34.98％.采用紫外光谱(UV)、荧光光谱(FS)、傅里叶红外光谱(FT-IR)、扫描电镜

(SEM)对牛骨胶原蛋白及其产物进行结构特性分析.通过UV分析表明BSC酶的降

解作用破坏了牛骨胶原蛋白的三股螺旋结构,游离出大量的氨基酸;FS分析表明牛骨

胶原蛋白分子表面CO、CONH2、COOH逐步增多,胶原蛋白肽中生色基团分布也

随之发生变化;FT-IR分析可知牛骨胶原蛋白经降解后的胶原蛋白肽的肽链上-

NH2+-排斥作用逐渐减弱,主要以β-转角为主;SEM表明酶破坏了牛骨胶原蛋白的

表面分子结构,使其变得疏松.%Inthiswork,theenzymatichydrolysisofbovine

bonecollagenwasstudiedbypre-purifiedbone-specificcollagenase

(BSC).Asevaluatedintermsofdegreeofhydrolysis.Theenzymatic

hydrolysisprocesswasstudiedthroughsinglefactorandquadraticgeneral

rotaryunitizeddesignexperiments.Theresultsshowedthattheoptimum

hydrolysisconditionswereasfollows:reactiontemperature46℃,the

amountofbovinebonecollagen5.14g/100mL,theamountofBSC

enzyme0.42g/100mL,reactiontime6h,pH6.5.Undertheseconditions,the

degreeofhydrolysisvaluewas34.98％.Thestructuralpropertiesofbovine

bonecollagenanditshydrolysatewerecharacterizedby

ultraviolet,fluorescence,fouriertransforminfraredspectroscopy,and

differentialscanningcalorimetry.TheUVanalysisshowedthattheBSC

degradationenzymedamagethebonecollagenthreehelixstructure,a

largenumberoffreeaminoacid;FSanalysisshowedthatthecollagen

molecularsurfaceC-O,CONH2andCOOHgraduallyincreased,collagen

peptidechromophoredistributionalsochanges;FT-IRanalysisshowedthat

thedegradationofcollagenfrombovinebonecollagenpeptideafterthe

thepeptidechainof-NH2+-repulsioneffectgraduallyweakened,mainlyin

thebetaangle;SEMshowedthattheenzymedamagedthesurfaceof

bovinecollagenmolecularstructure,andmadeitloose.

【期刊名称】《食品与机械》

【年(卷),期】2017(033)007

【总页数】7页(P40-46)

【关键词】牛骨;胶原蛋白;酶解;结构特性

【作者】刘丽莉;李丹;杨陈柳;孟圆圆;代晓凝

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