哈茨木霉产β-1,3-葡聚糖内切酶的发酵工艺条件研究  

Study on liquid fermentation conditions for Trichoderma harzianum to produce β-1,3-endoglucanase

作　　者：李珊[1] 詹晓北[1,2] 郑志永[1] 朱莉 李晶[1] 徐敏[1] 

机构地区：[1]江南大学生物工程学院糖化学与生物技术教育部重点实验室,江苏无锡214122 [2]江苏瑞光生物科技有限公司,江苏无锡214125

出　　处：《工业微生物》2015年第2期39-46,共8页Industrial Microbiology

基　　金：国家自然科学基金项目(31171640;31271888);国家高技术发展计划(863)项目(2012AA021505);无锡市科技支撑计划(农业;CLE01N1208);无锡市130计划

摘　　要：对哈茨木霉(Trichoderma harzianum GIM 3.442)产β-1,3-葡聚糖内切酶的液体发酵条件进行了单因素优化实验,确定了最佳培养基成分和培养条件,在此基础上通过正交试验设计对复合碳源(葡萄糖、茯苓多糖)、胰蛋白胨、Na NO3和磷酸盐进行了L9(34)试验,研究了4种因素对哈茨木霉产酶的影响,确定了最佳培养条件:葡萄糖42.0 g/L,茯苓多糖18.0 g/L,胰蛋白胨15.0 g/L,Na NO35.0 g/L,初始p H 6.0,接种量8%,28℃,110 r/min培养6 d。优化后总酶活Etotal和β-1,3-葡聚糖内切酶Eendo活力达到了471.6 U/m L和327.4 U/m L,比优化前分别提高了7.3倍和23倍,且内切酶占总酶活的比例Eendo/Etotal由0.24增加到0.71,效果显著。The liquid flask fermentation conditions including medium composition and cultural conditions of Trichoderma harzianum Rifai GIM 3. 442 for producing β-1,3-endoglucanase were optimized. Then the effects of four factors such as composite carbon （glucose,pachymaran）,tryptone,NaNO3 and KH2 PO4 were optimized by orthogonal experiment. Re-sults showed that the optimal medium conditions were as follows：glucose 42. 0 g/L,pachymaran 18. 0 g/L,tryptone 15. 0 g/L,NaNO3 5. 0 g/L;initial pH 6. 0,inoculum size 8%,at 28 °C,110 r/min and incubation for 6 d. The total enzyme activity Etotal and β-1,3-endoglucanase activity Eendo reached 471. 6 U/mL and 327. 4 U/mL,increased by 7. 3-fold and 23-fold respectively compared with those of the original activity,and the ratio of β-1,3-endoglucanase in total enzyme activity increased significantly from 0 . 24 to 0 . 71 .

关 键 词：β-1 3-葡聚糖内切酶 哈茨木霉 发酵优化 正交试验 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]