碱性蛋白酶和复合蛋白酶双酶水解羊骨粉的工艺研究

The Study on the Hydrolysis of Alcalase and Protamex to Sheep Bone Powder

[目的]碱性蛋白酶和复合蛋白酶为水解羊骨粉制备胶原多肽效果最好的两种单酶,为进一步提高酶解效果。[方法]以羊骨粉为原料,进行双酶水解,比较同步水解与顺序水解的效果,确定最优水解工艺。[结果]同步水解AB2组合的水解效果最优,各水解评价指标均显著高于单酶水解和顺序水解(P<0.05),与顺序水解相比不显著(P>0.05),但优于顺序水解且大大节省了时间,其水解工艺为:底物浓度9%,碱性蛋白酶和复合蛋白酶的加酶量分别为5.00%和4.25%,起始pH 8.5,50℃水解5h时,水解度达29.79%,氨基酸态氮28.13%,多肽生成量110.43mg·g-1,短肽得率73.88%。[结论]双酶同步水解羊骨粉提高酶解效果的同时,也为充分水解羊骨制备胶原多肽提供了试验支持。

[Objective]Alcalase and protamex were the best enzymes for collagen polypeptide that hydrolyzed from the sheep bone through screening.[Methods]This research compared the effect of sequentially and simultaneously hydrolyzing to further improve the enzyme effect and to determine the best condition for hydrolyzing.[Results]This experiment showed that：The group of simultaneously hydrolyzing AB2 that the hydrolysis of indicators were extremely higher than the single（P〈0.05）compared with sequentially hydrolyzing is insignificant（P〉0.05）but saved time extremely in condition of 9% substrate,5%（E/S）alcalase and 4.25%（E/S）protamex,initial pH 8.5,50 ℃,5h.From researching that the degree of hydrolysis,amount of amino-acid nitrogen,amount of formed polypeptides,the obtaining rate of oligopeptides partly researched to 29.79%,28.13%,110.43 mg·g^-1 and 73.88%.[Conclusion]To sum up,double enzyme simultaneously hydrolyzing not only could improve the enzyme effect but also could provide experimental support for the preparation of collagen polypeptide from sheep bone.