耐高温β-环状糊精葡萄糖基转移酶高产菌株XW-6-66的选育及发酵条件的研究  

A research on screening and fermentation conditions of thermostable β-Cyclodextrin Glucanotransferase high-production strain XW-66

作　　者：孟艳芬[1] 许波[1] 高润池[1] 货胜英 王晓燕[1] 黄遵锡[1] 

机构地区：[1]云南师范大学生命科学学院,生物能源持续开发利用教育部工程研究中心,云南师范大学酶工程重点实验室,昆明650092

出　　处：《食品科技》2009年第5期10-15,共6页Food Science and Technology

基　　金：云南省自然科学基金(2005C0018Q);云南省应用基金重点项目(2006C0004Z)

摘　　要：从云南宣威地区的淀粉厂周围土壤中筛选到一株热稳定性能好的β-环状糊精葡萄糖基转移酶高产菌XW-6-66并对其发酵条件进行研究,该菌株发酵72h在pH9.0、70℃条件下发酵液酶活为2173.33U/mL,酶学特性非常适合于环状糊精的工业化生产。通过分子生物学及其生理生化鉴定该菌株为类芽孢杆菌(Paenibacillus sp.)。此外,分别对该菌发酵培养基的碳源、氮源及pH进行了单因子分析,并对这3个因素进行了正交实验及验证,最终确定该菌的最佳发酵培养基配方为:0.3%可溶性淀粉,1.0%牛肉浸膏,1%蛋白胨,0.2%Na2CO3,0.13%K2HPO4·3H2O,0.02%MgSO4·7H2O,pH为9.0。在该条件下,菌株XW-6-66产β-CGTase的酶活由优化前2173.33U/mL提高到3622.94U/mL。A thermostable β-cyclodextdn glucanotransferase high-production strain XW-6-66 was isolated from soil of the amylum factoy Xuanwei Yunan. The experiment is a tentative research on the fermentation conditions and enzymatic properties with the object of the high-yield strain XW-6-66 with the production of β- Cyclodextrin Glucanotransferase. The enzyme activity was 2173.33 U/mL at 72 h With a most proper temperature of 70 ℃, and a most proper pH of 9.0, its enzymatic properties are rather applicable to the industrialization of cyclodextrin. The strain was identified as Paenibacillus sp.by the methods of molecular biology and physiological and biochemical characters. In addition, the experiment carries on a one-way analysis of the carbon source, nitrogen source and pH of fermentation medium, and orthogonal experiments and tests go to the three factors to finally define the best ingredient of the bacteria culture medium： 0.3% starch soluble, 1.0% beef extract, 1% tryptone, 0.2% Na2CO3, 0.13% K^PO4.3H20, 0.02% MgSO4.7H20, pH9.0. Under these conditions, the promotion of the β-CGTase activity of a strain XW-6-66 is shown by from the pre-optimized data 2173.33 Unit per micro-liter to 3622.94 Unit per micro-liter.

关 键 词：环状糊精葡萄糖基转移酶 Β-环状糊精 菌种选育 发酵条件 

分 类 号：TS201.3[轻工技术与工程—食品科学]