谷氨酰胺转氨酶发酵工艺优化及其酶学性质
Fermentation Process Optimization of Transglutaminase and Its Enzymatic Properties
王兴吉 1刘文龙 1郭庆亮 1张杰1
作者信息
- 1. 山东隆科特酶制剂有限公司,山东省酶制剂生物发酵技术重点实验室,山东临沂276400
- 折叠
摘要
[目的]研究茂源链霉菌LD195产TGase的最佳发酵工艺条件,并探讨TGase的酶学特性.[方法]采用L9(34)正交试验对产谷氨酰胺转氨酶的茂源链霉菌菌株培养基和培养条件进行优化.[结果]最佳无机盐配方为Na2 HPO42.0 g/L,MgSO4·7H2O 1.5 g/L,KH2PO4 2.5 g/L,CaCl2 3.0 g/L,最佳培养条件为培养温度30℃,装液量25mL,摇床转速200 r/min,接种量10％.该谷氨酰胺转氨酶的最适反应温度为50℃,在40～60℃条件下保温30 min酶活基本没有损失;最适反应pH为6.0,在pH为5.0 ～7.0条件下保存30 min仍具有85％以上酶活;同时Fe3+、Cu2+、Zn2+会强烈抑制酶的活性.[结论]茂源链霉菌LD195产TGase具有良好的耐酸性和耐热性,在商业化生产中具有广阔前景.
Abstract
[Objective]To investigate the optimum fermentation conditions of TGase produced by Streptomyces lanceolata LD195 and explore the enzymatic properties of TGase.[Method] L9(34) orthogonal design on media and culture conditions of Streptomyces lanceolata were optimized.[Result]The best medium:Na2HPO4 2.0 g/L,MgSO4 · 7H2O 1.5 g/L,KH2PO4 2.5 g/L,CaCl2 3.0 g/L.The optimal culture conditions:culture temperature 30 ℃,bottling capacity 25 mL,agitation rate 200 r/min,inoculums size 10％.The produced TGase exhibited optimum activity at 50 ℃ and pH 6.0.It was stably incubated in 40 ～60 ℃ for 30 min and the remained enzyme activity was above 85％ dealed with pH 5.0 ～7.0 for 30 min.The enzyme was strongly inactivated by Fe3+,Cu2+,Zn2+.[Conclusion] TGase produced by Streptomyces lanceolata LD195 has good acid resistance and heat resistance,and which has broad prospects in commercial production.
关键词
培养基/茂源链霉菌/谷氨酰胺转氨酶/酶学性质Key words
Medium/Streptomyces lanceolata/Transglutaminase/Enzymatic properties引用本文复制引用
基金项目
山东省科技重大专项(2014CGZH1305)
出版年
2017
国家科技期刊平台
NSTL
维普
万方数据