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/100mL、BSC添加量0.42g/100 mL、反应时间6h、pH 6.5,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C═O、CONH_2、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—NH_2^+—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。
In this work,the enzymatic hydrolysis of bovine bone collagen was studied by pre-purified bone-specific collagenase（BSC）.As evaluated in terms of degree of hydrolysis.The enzymatic hydrolysis process was studied through single factor and quadratic general rotary unitized design experiments.The results showed that the optimum hydrolysis conditions were as follows：reaction temperature 46 ℃,the amount of bovine bone collagen 5.14g/100 mL,the amount of BSC enzyme 0.42g/100 mL,reaction time 6h,pH 6.5.Under these conditions,the degree of hydrolysis value was 34.98%.The structural properties of bovine bone collagen and its hydrolysate were characterized by ultraviolet,fluorescence,fourier transform infrared spectroscopy,and differential scanning calorimetry.The UV analysis showed that the BSC degradation enzyme damage the bone collagen three helix structure,a large number of free amino acid;FS analysis showed that the collagen molecular surface C ═ O,CONH_2 and COOH gradually increased, collagen peptide chromophore distribution also changes;FT-IR analysis showed that the degradation of collagen from bovine bone collagen peptide after the the peptide chain of—NH_2~＋— repulsion effect gradually weakened,mainly in the beta angle;SEM showed that the enzyme damaged the surface of bovine collagen molecular structure,and made it loose.
出处
《食品与机械》
CSCD
北大核心
2017年第7期40-46,共7页
Food and Machinery
基金
国家自然科学基金(编号:31401622)
公益性行业(农业)科研专项(编号:201303084)
河南省重点攻关项目(编号:152102110080)
河南省教育厅自然科学研究项目(编号:13A550255)
河南省重大专项(编号:161100110900)
关键词
牛骨
胶原蛋白
酶解
结构特性
novel collagenase
bovine bone collagen
enzymatic hydrolysis
optimization
structural properties
