[1]郑双凤,谭武贵,丰来,等.枯草芽孢杆菌NTGB-178高产芽孢发酵工艺优化[J].南京农业大学学报,2017,40(6):1031-1040.[doi:10.7685/jnau.201702023]
 ZHENG Shuangfeng,TAN Wugui,FENG Lai,et al.Optimization of sporulation fermentation process of Bacillus subtilis NTGB-178[J].Journal of Nanjing Agricultural University,2017,40(6):1031-1040.[doi:10.7685/jnau.201702023]
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枯草芽孢杆菌NTGB-178高产芽孢发酵工艺优化()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
40卷
期数:
2017年6期
页码:
1031-1040
栏目:
出版日期:
2017-11-10

文章信息/Info

Title:
Optimization of sporulation fermentation process of Bacillus subtilis NTGB-178
作者:
郑双凤 谭武贵 丰来 罗志威 徐滔明 张冬雪 谭石勇
农业部植物营养与生物肥料重点实验室/湖南泰谷生物科技股份有限公司, 湖南 长沙 410205
Author(s):
ZHENG Shuangfeng TAN Wugui FENG Lai LUO Zhiwei XU Taoming ZHANG Dongxue TAN Shiyong
Key Laboratory of Plant Nutrition and Biological Fertilizer, Ministry of Agriculture/Hunan Taigu Bio-Tech Co. Ltd., Changsha 410205, China
关键词:
枯草芽孢杆菌优化芽孢响应面法发酵工艺
Keywords:
Bacillus subtilisoptimizationsporeresponse surface methodologyfermentation technology
分类号:
Q939.9
DOI:
10.7685/jnau.201702023
摘要:
[目的] 优化枯草芽孢杆菌NTGB-178发酵工艺,为提高发酵液中的生物量与芽孢产量,降低该菌产品制剂生产成本提供技术支撑。[方法] 采用响应面法(RSM)对NTGB-178发酵工艺进行优化。在单因素试验基础上,利用Plackett-Burman设计试验,筛选出影响NTGB-178芽孢产量的主要影响因子。利用最陡爬坡试验,确定逼近存在最大响应值的中心点。最后利用Design-Expert软件设计Box-Behnken试验,对其结果进行多元二次回归拟合,建立响应面方程,绘制响应面图,并对影响发酵产孢的主要因素及其交互作用进行响应面分析和评价,确定最优的液体发酵培养条件。[结果] 麸皮、NaCl、初始pH为影响NTGB-178芽孢产量的主要因素,方程预测最优水平为:麸皮10.35 g·L-1,NaCl 4.41 g·L-1,初始pH6.07,其他条件分别为:玉米粉12.5 g·L-1,豆粕25.0 g·L-1,CaCO31.0 g·L-1,MgSO4·7H2O 2.0 g·L-1,接种量4%,装液量65 mL/250 mL,培养温度36℃,摇床转速220 r·min-1,培养时间36 h。经验证,优化后发酵液中生物量为6.55×109 CFU·mL-1,芽孢产量为6.16×109 CFU·mL-1,芽孢产量较优化前提高了8.48倍。最后在中试培养条件下,发酵36 h后芽孢产量最高为7.33×109 CFU·mL-1,验证了摇瓶优化条件的稳定性。[结论] 采用单因素试验与响应面法相结合的方法优化发酵工艺,可显著提高NTGB-178的生物量与芽孢产量,此发酵工艺为芽孢杆菌NTGB-178工业化扩大生产奠定了基础。
Abstract:
[Objectives] The objective of this study is to optimize the fermentation conditions of Bacillus subtilis NTGB-178 for improving the liquid fermentation of biomass and the spore yield, then provide a cost-effective fermentation technique.[Methods] Response surface methodology (RSM)was used to optimize the fermentation conditions. The main factors which affected the liquid fermentation were screened by Plackett-Burman design, based on the single factor experiment. The steepest ascent path was used to identify the central point of response surface and approach the optimal region. The Box-Behnken design was adopted to identify the optimal fermentation conditions by establishing a multiple quadratic regression equation, analyzing and evaluating the factors which affect the spore yield of B. subtilis NTGB-178.[Results] Bran, NaCl and initial pH were identified as the main factors. The optimum conditions, which were predicted by the response surface equation, were bran 10.35 g·L-1, NaCl 4.41 g·L-1, and initial pH6.07. The other factors were as follows:corn flour 12.5 g·L-1, bean pulp 25.0 g·L-1, CaCO31.0 g·L-1, MgSO4·7H2O 2.0 g·L-1, inoculation amount 4%, loaded liquid volume 65 mL/250 mL, rotation speed 220 r·min-1and cultivating at 36℃ for 36 h. After verification, the biomass reached 6.55×109 CFU·mL-1, and the spore yield reached 6.16×109 CFU·mL-1, which was improved 8.48 times compared to the result before optimization. In the 100 L fermenter enlarge cultivations, the spore yield was 7.33×109 CFU·mL-1. The stability of the optimal fermentation conditions wase indicated.[Conclusions] Single factor experiment and RSM were simultaneously used in the optimization of fermentation of B. subtilis NTGB-178 for the biomass and spore production, which was proved to be effective. This study laid the foundation for industrialized production of B. subtilis NTGB-178.

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备注/Memo

备注/Memo:
收稿日期:2017-02-20。
基金项目:国家重点研发计划项目(2016YFD0300904-2);湖南省重点研发计划项目(2016NK2016)
作者简介:郑双凤,硕士研究生,助理工程师。
通信作者:谭石勇,博士,研究方向为植物营养与生物肥料,E-mail:Shiy-tan@hotmail.com。
更新日期/Last Update: 1900-01-01