基于核糖体工程理论的常压室温等离子体诱变筛选多杀菌素高产菌

doi:10.13523/j.cb.20140111

中国生物工程杂志

2014, Vol. 34

Issue (1): 71-78 DOI: 10.13523/j.cb.20140111

技术与方法

基于核糖体工程理论的常压室温等离子体诱变筛选多杀菌素高产菌

乔长晟^1,2, 赵男¹, 石漫漫¹, 朱明¹, 李雪²

1. 天津科技大学工业发酵微生物教育部重点实验室天津 300457;
2. 天津北洋百川生物技术有限公司天津 300457

Mutation Screening of Production Strains of Spinosad Based on Ribosome Engineering Theory through ARTP

QIAO Chang-sheng^1,2, ZHAO Nan¹, SHI Man-man¹, ZHU Ming¹, LI Xue²

1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, China;
2. Tianjin Peiyang Biotrans Co., Ltd., Tianjin 300457, China

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摘要： 利用核糖体工程理论，采用常压室温等离子体（ARTP）诱变对刺糖多孢菌（Saccharopolyspora spinosa）进行诱变筛选。以刺糖多孢菌QYLZ 88912菌株为出发菌株，对其进行ARTP诱变，选取三个不同致死率的照射时间，将孢子悬液混合，以增加抗性筛选几率。然后基于核糖体工程技术理论对混合孢子悬液进行初筛，最终筛选出了1株多杀菌素高产菌Sg²⁰⁰Rif¹¹⁰St⁴⁰Er⁹⁰-028。该菌株同时具有磺胺胍、利福平、链霉素、红霉素的多重抗性，摇瓶发酵试验表明，发酵7d后多杀菌素浓度可达到1 516.93 mg/L，较出发菌株QYLZ 88912的产量610.75 mg/L提高了148.37%，且遗传性稳定。以得到的高产菌Sg²⁰⁰Rif¹¹⁰St⁴⁰Er⁹⁰-028作为出发菌株，对发酵培养基进行响应面优化实验，优化后的培养基（g/L）：葡萄糖46.97、麦芽糊精35、酵母粉40.36、水浸棉籽粉32.88、碳酸钙3，多杀菌素的产量为2 361.81 mg/L，比出发菌株的产量提高了286.71%。

关键词： 多杀菌素; 核糖体工程; 常压室温等离子体诱变

Abstract: Based on the ribosome engineering theory, the atmospheric and room temperature plasma (ARTP) was used in screening of Saccharopolyspora spinosa. Taking the strain Saccharopolyspora spinosa QYLZ 88912 as the original strain, choose the spore suspension of three different irradiation time after ARTP mutation, then mix them to increase the resistance screening probability. A high-yield strain S.spinosa Sg²⁰⁰Rif¹¹⁰St⁴⁰Er⁹⁰-028 was obtained after screening to the mixed spore suspension. This strain had the resistance of sulfaguanidine, rifampin, streptomycin, erythromycin. Its production of spinosad reached 1516.93 mg/L under the level of shake flask fermentation through 7 days, which was 148.37% higher than that of the original strain S.spinosa QYLZ 88912. The subculture experiments indicated that the hereditary characteristic of high productivity of S.spinosa Sg²⁰⁰Rif¹¹⁰St⁴⁰Er⁹⁰-028 was relatively stable. The fermentation materials of spinosad producing strain were optimized by the response surface methodology based on the single factor experiment. Under the optimized medium, the spinosad was increased by 286.71% compared with the previous condition.

Key words: Atmospheric and room temperature plasma (ARTP) Spinosad Rribosome engineering

收稿日期: 2013-10-24 出版日期: 2014-01-25

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通讯作者: 赵男，E-mail：791692024@qq.com E-mail: 791692024@qq.com

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引用本文:

乔长晟, 赵男, 石漫漫, 朱明, 李雪. 基于核糖体工程理论的常压室温等离子体诱变筛选多杀菌素高产菌[J]. 中国生物工程杂志, 2014, 34(1): 71-78.

QIAO Chang-sheng, ZHAO Nan, SHI Man-man, ZHU Ming, LI Xue. Mutation Screening of Production Strains of Spinosad Based on Ribosome Engineering Theory through ARTP. China Biotechnology, 2014, 34(1): 71-78.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140111 或 https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I1/71

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