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暹罗芽孢杆菌高产γ-聚谷氨酸的发酵条件优化
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国家重点研发计划(2018YFD0400404)


Optimization of fermentation conditions for poly-γ-glutamic acid production by Bacillus siamensis
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    摘要:

    【背景】γ-聚谷氨酸(poly-γ-glutamic acid,γ-PGA)产生菌多为枯草芽孢杆菌(Bacillus subtilis)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、地衣芽孢杆菌(Bacillus licheniformis)等,而暹罗芽孢杆菌(Bacillus siamensis)相关研究较少。【目的】研究暹罗芽孢杆菌产γ-PGA的液体发酵条件。【方法】以自行分离的暹罗芽孢杆菌CAU83为出发菌株进行液体发酵,通过单因素试验和正交试验法研究了碳氮源、前体物质、发酵温度及pH对菌株生产γ-PGA的影响。【结果】经摇瓶优化,γ-PGA的最适碳源、氮源和前体物质分别为乳糖30 g/L、酵母提取物5 g/L和L-谷氨酸钠60 g/L,最适培养条件为发酵温度37 ℃和pH 7.0,γ-PGA产量由8.4 g/L提升至30.1 g/L,比优化前提高了260%。经分批补料发酵,60 h时γ-PGA产量最高为59.5 g/L,比摇瓶提高了98%,产率为0.99 g/(L·h)。所产γ-PGA分子量为3.8×106 Da,聚合度较高。【结论】确定了暹罗芽孢杆菌CAU83产γ-PGA的最适发酵条件,为该菌株的工业化生产和应用提供了依据。

    Abstract:

    [Background] The commonly known poly-γ-glutamic acid (γ-PGA) producers are Bacillus subtilis, B. amyloliquefaciens, and B. licheniformis, while little is known about B. siamensis. [Objective] To study liquid fermentation conditions for γ-PGA production by B. siamensis. [Methods] B. siamensis CAU83, isolated by our laboratory, was used to produce γ-PGA by liquid fermentation. The effects of carbon sources, nitrogen sources, precursors, temperature, and pH on the synthesis of γ-PGA in shake flasks were investigated by single factor test and orthogonal design. [Results] The optimal carbon source, nitrogen source, and precursor for the synthesis of γ-PGA were 30 g/L lactose, 5 g/L yeast extract, and 60 g/L L-sodium glutamate, respectively. The optimal fermentation conditions were 37 ℃ and pH 7.0. The yield of γ-PGA increased by 260% from 8.4 g/L before optimization to 30.1 g/L after optimization. The fed-batch fermentation showed the peak yield (59.5 g/L) of γ-PGA at the time point of 60 h, with a productivity of 0.99 g/(L·h), which increased by 98% compared with the yield in shake flasks. The produced γ-PGA had a molecular weight of 3.8×106 Da and a high polymerization degree. [Conclusion] The optimal fermentation conditions for B. siamensis CAU83 producing γ-PGA determined in this study provide a basis for the industrial production and application of this strain.

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林格儿,刘宏,刘海杰,闫巧娟,江正强. 暹罗芽孢杆菌高产γ-聚谷氨酸的发酵条件优化[J]. 微生物学通报, 2022, 49(8): 3335-3345

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  • 收稿日期:2021-11-30
  • 最后修改日期:
  • 录用日期:2022-01-17
  • 在线发布日期: 2022-07-28
  • 出版日期: 2022-08-20
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