N+注入诱变选育β-环状糊精葡萄糖基转移酶高产菌株及其发酵条件优化

食品科学 ›› 2012, Vol. 33 ›› Issue (15): 239-245.

N+注入诱变选育β-环状糊精葡萄糖基转移酶高产菌株及其发酵条件优化

张洪斌，凌国庆，胡雪芹，凡宁

合肥工业大学制药工程系

收稿日期:2012-03-23 修回日期:2012-07-20 出版日期:2012-08-15 发布日期:2012-09-07
通讯作者: 张洪斌 E-mail:zhb5678@163.com
基金资助:
安徽省长三角科技联合攻关项目;合肥工业大学大学生创新性实验计划项目

Screening of a High-Yield β-Cyclodextrin Glycosyltransferase-Producing Bacillus cereus Strain and Optimization of Its Fermentation Conditions

Received:2012-03-23 Revised:2012-07-20 Online:2012-08-15 Published:2012-09-07
Contact: zhang hong-bin E-mail:zhb5678@163.com

摘要/Abstract

摘要： 以一株产β-环状糊精葡萄糖基转移酶的蜡状芽孢杆菌BC-0801为出发菌株，利用10keV，剂量为40×2.5×1013 ions/cm2氮离子进行诱变选育，获得一株高产β-环状糊精葡萄糖基转移酶的突变体BC-0802。对该突变体BC-0802发酵培养基的碳源、氮源及无机离子进行单因素试验分析，采用3个因素正交试验确定其最佳发酵培养基成分质量浓度及发酵条件为：玉米粉30g/L、酵母浸膏15g/L、K2HPO4 1g/L，发酵温度30℃、pH 9.0、接种量2%、发酵周期36h。结果表明；突变体的平均酶活力可达3415.8U/mL，相较于出发菌株酶活力提高了3.67倍，且该突变体的遗传稳定性较好。

关键词: N+注入, β-环状糊精葡萄糖基转移酶, 筛选, 诱变, 工艺优化

Abstract: Bacillus cereus BC-0801, a β-cyclodextrin glycosyltransferase (β-CGTase)-producing strain, was mutagenized by means of low energy (10 keV) N+ ion implantation at a dose of 40 × 2.5 × 1013 ions/cm2 to obtain a high-yield mutant strain designated as BC-0802. The effects of carbon and nitrogen sources and inorganic salt (K2HPO4) in the medium on β-CGTase production by BC-0802 were investigated by one-factor-at-a-time method. The three components were optimized by orthogonal array design method. The optimal fermentation medium was composed of corn flour 30 g/L, yeast extract 15 g/L, K2HPO4 1 g/L. The optimal fermentation conditions were determined as inoculum size 2%, fermentation period 36 h, temperature 30 ℃, and pH 9.0. The average β-CGTase activity of the mutant strain was 3415.8 U/mL (n = 5), which was 3.67 times higher than that of the starting strain. Moreover the mutant strain showed good genetic stability.

Key words: N+ ion implantation, β-cyclodextrin glycosyltransferase, screening, mutagenesis, technological optimization

中图分类号:

Q936

张洪斌，凌国庆，胡雪芹，凡宁. N+注入诱变选育β-环状糊精葡萄糖基转移酶高产菌株及其发酵条件优化[J]. 食品科学, 2012, 33(15): 239-245.

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