天然产物研究与开发 ›› 2019, Vol. 31 ›› Issue (8): 1425-1433.doi: 10.16333/j.1001-6880.2019.8.018

• 开发研究 • 上一篇    下一篇

高效表达淀粉酶Bacillus koreensis的培养基响应面优化

何伟#,李菁菁#,包可翔,颜奕华,姜振锟,林俭,陈善义*,李易非*   

  1. 福建中烟工业有限责任公司技术中心,厦门 361021
  • 出版日期:2019-09-10 发布日期:2019-09-10
  • 基金资助:

    福建中烟工业有限责任公司技术中心科技项目(JSZ XKJJH2016001)

Optimization of culture medium for a high performance amylase-producing Bacillus koreensis by response surface methodology

HE Wei#,LI Jing-jing#,BAO Ke-xiang,YAN Yi-hua,JIANG Zhen-kun,LIN Jian,CHEN Shan-yi*,LI Yi-fei*   

  1. Technology Center,Fujian Tobacco Industry Co.,Ltd.,Xiamen 361021,China
  • Online:2019-09-10 Published:2019-09-10

摘要: 为降低烟叶中的淀粉含量,提高烟叶的可用性,从云南马龙C3F-2014烟叶表面筛选出一株高产淀粉酶的细菌,经16sRNA测序鉴定为Bacillus koreensis。本研究利用响应面法优化Bacillus koreensis的培养基提高了其表达淀粉酶的产量。首先,单因素优化试验表明培养基的最优碳源、氮源和金属离子分别为淀粉、蛋白胨和Ca2+,培养条件单因素试验表明Bacillus koreensis最适初始pH、最适温度和最适接种量分别为pH8.0、37 ℃和3%。利用Box-Behnken中心组合设计对可溶性淀粉、蛋白胨、Ca2+设计三因素三水平实验,通过响应面回归分析,得到模型预测的最优培养基条件。在18.74 g/L可溶性淀粉,21.56 g/L蛋白胨,0.52 g/L CaCl2的培养基条件下Bacillus koreensis产淀粉酶最高。验证试验得到的淀粉酶活力达到959.39±22.34 U/mL,与模型的预测值相近,比未优化前提高了69.76%。本研究结果为烟草天然源淀粉酶处理烟叶,提高烟叶品质的工业化应用提供了基础。

关键词: Bacillus koreensis, 淀粉酶, 培养基优化, 响应面, 烟叶

Abstract: In order to reduce the content of starch in tobacco leaves and improve the usability of tobacco leaves,A high performance amylase-producing strain was isolated from tobacco leaf samples C3F from Malong which is in Yunnan province,China.It was identified as Bacillus koreensis by 16s RNA analysis.The culture medium of Bacillus koreensis was optimized by response surface methodology,remarkable elevated the expression of amylase.First,single parameter experiments show that the optimal carbon,nitrogen,metal ion, initial pH,temperature and inoculums size were starch,peptone,Ca2+ ,8.0,37 ℃,3% respectively.Then,a three factors-three levels experiment was carried out based on the Box-Behnken central composite design.The highest amylase activity was 959.39±22.34 U/mL in the validation experiment under the optimized conditions of 18.74 g/L soluble starch,21.56 g/L peptone,0.52 g/L CaCl2.The amylase activity of Bacillus koreensis was elevated 69.76% than before,which was similar to the predicted value of the model.This study was based on Bacillus koreensis from natural tobacco leaves,providing a basis for industrial application of improving tobacco quality.

Key words: Bacillus koreensis, amylase, medium culture optimization, response surface methodology, tobacco

中图分类号: 

R284.2 Q556