食品与发酵工业 >

2021 , Vol. 47 >Issue 4: 136 - 143

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.025196

生产与科研应用

拟蕈状芽孢杆菌Gxun-30产角蛋白酶液体发酵条件优化

  • 张红岩 ,
  • 张妮 ,
  • 杨梦莹 ,
  • 刘聪 ,
  • 杨立芳 ,
  • 申乃坤 ,
  • 姜明国
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  • 1(广西民族大学 海洋与生物技术学院,广西多糖材料与改性重点实验室,广西高效微生物与植物资源利用重点实验室,广西 南宁,530006)
    2(广西民族大学 化学化工学院,广西林产化学与工程重点实验室,广西 南宁,530006)
高级工程师(申乃坤教授为通讯作者,E-mail:shennaik05@126.com)

收稿日期: 2020-07-28

  修回日期: 2020-09-01

  网络出版日期: 2021-03-16

基金资助

国家自然科学基金(31660022;31660005);广西科技重点研发计划(AA18242026);广西自然科学基金(2018GXNSFAA28113;2019GXNSFAA185003);广西科技基地与人才专项(2017AD19029;AD18281066);广西民族大学相思湖青年学者创新团队(2017-6);广西民族大学科学研究项目(2018KJQD17)

收起

Optimization of liquid fermentation conditions of keratinase produced by Bacillus paramycoides Gxun-30

  • ZHANG Hongyan ,
  • ZHANG Ni ,
  • YANG Mengying ,
  • LIU Cong ,
  • YANG Lifang ,
  • SHEN Naikun ,
  • JIANG Mingguo
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  • 1(Guangxi Key Laboratory for Polysaccharide Materials and Modifications, Guangxi Key Laboratory of Microbial plant Resources and Utilization, School of Marine Sciences and Biotechnology, Guangxi University for Nationalities, Nanning 530006, China)
    2(Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, School of Chemistry and Chemical Engineering, Guangxi University for Nationalities, Nanning 530006, China)

Received date: 2020-07-28

  Revised date: 2020-09-01

  Online published: 2021-03-16

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摘要

为提高海洋来源拟蕈状芽孢杆菌(Bacillus paramycoides) Gxun-30产角蛋白酶的能力,该文利用单因素及响应面法对该菌产酶的培养基和培养条件进行了优化。先利用单因素试验对羽毛浓度、碳源、氮源、无机盐、初始pH值、发酵时间及接种量等影响菌株产酶条件进行了优化。结果表明,羽毛15 g/L、果糖10 g/L、玉米浆4.0 g/L、初始pH 6.5、氯化钙0.15 g/L、接种量 2.0%、接种发酵48 h 后酶活达到最高。再利用Plackett-Burman试验确定对菌株产酶有显著影响的3个因素为玉米浆、氯化钙及羽毛浓度;结合最陡爬坡及响应面试验优化方法对这3个显著因素进行优化,获得最优产酶条件为玉米浆8.17 g/L,氯化钙0.27 g/L,羽毛含量13.58 g/L,在此发酵条件下,模型预测角蛋白酶酶活为1 866.47 U/mL,验证试验实测值达到1 810.98 U/mL,较优化前酶活227.38 U/mL提高了7.96倍。

关键词： 拟蕈状芽孢杆菌; 角蛋白酶; 羽毛降解; 响应面优化

本文引用格式

张红岩 , 张妮 , 杨梦莹 , 刘聪 , 杨立芳 , 申乃坤 , 姜明国 . 拟蕈状芽孢杆菌Gxun-30产角蛋白酶液体发酵条件优化[J]. 食品与发酵工业, 2021 , 47(4) : 136 -143 . DOI: 10.13995/j.cnki.11-1802/ts.025196

Abstract

In order to enhance the keratinase activity of marine sourced Bacillus paramycoides Gxun-30, the liquid culture medium and conditions were optimized by single factor experiment and response surface method. First, the effects of feathers concentration, carbon source, nitrogen source, inorganic salts, initial pH, fermentation time and inoculation amount on the keratinase production were investigated by single factor experiments. The optimal results corresponding to each single factor were: feathers concentration 15 g/L, fructose 10 g/L, corn steep liquor 4.0 g/L, initial pH 6.5, CaCl2 0.15 g/L, inoculation volume 2.0%, fermentation time 48 h. Secondly, using Plackett-Burman experiment, the main significant factors were identified as corn steep liquor, CaCl2 and feathers amount. Finally, the optimal levels of these three factors were determined with the steepest ascent experiment and response surface method. The optimal conditions for enzyme production were: corn steep liquor 8.17 g/L, CaCl2 0.27 g/L, and feathers amount 13.58 g/L. Under the optimized conditions, the keratinase activity was predicted as 1 866.47 U/mL. The measured keratinase activity reached 1 810.98 U/mL, which was 7.96 times of the initial keratinase activity 227.38 U/mL.

Key words： Bacillus paramycoides; keratinase; feather-degrading; optimization; response surface methodology

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