Mutation of Cl.beijerinckii protoplasts by <sup>60</sup>Co-γ irradiation

LIU Ya; LIU Hongjuan; ZHANG Jian'an; CHENG Keke; CHEN Zongdao

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Mutation of Cl.beijerinckii protoplasts by ⁶⁰Co-γ irradiation

更新时间：2025-12-10

- Mutation of Cl.beijerinckii protoplasts by ⁶⁰Co-γ irradiation
- CIESC Journal Issue 10, Pages: 2549-2554(2009)
- 作者机构：
  
  1. 西南大学食品学院
  2. 清华大学核能与新能源技术研究院
  3. 石河子大学食品学院
- 作者简介：
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- Online First：05 October 2009，
  
  Published：05 October 2009
- 稿件说明：
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LIU Ya, LIU Hongjuan, ZHANG Jian'an, et al. Mutation of Cl.beijerinckii protoplasts by ⁶⁰Co-γ irradiation[J]. CIESC Journal, 2009, (10): 2549-2554.
DOI：

LIU Ya, LIU Hongjuan, ZHANG Jian'an, et al. Mutation of Cl.beijerinckii protoplasts by ⁶⁰Co-γ irradiation[J]. CIESC Journal, 2009, (10): 2549-2554. DOI：

摘要

以产丁醇菌Cl.beijerinckii为出发菌株，将其原生质体化后进行60Co-γ辐照诱变，筛选出了一株稳定高产的变异株。原生质体的最优制备条件为：将培养至15 h的细胞添加1 g.L-1甘氨酸和0.3 U.L-1青霉素培养2 h后，37℃下，2×104 U.L-1溶菌酶作用8 h脱壁。经平板计数，原生质体形成率可达99.82％，原生质体再生率达10.12％。采用60Co-γ射线进行辐照诱变，当辐照剂量率5 Gy.min-1，辐照剂量为500 Gy时，原生质体致死率可达99.3％。经反复筛选，获得一株丁醇高产菌株R3，该菌具有较高的遗传稳定性。在P2半合成液体培养基中，37℃下静置培养72 h，总溶剂产量较出发菌株提高81.46％，丁醇产量提高85.41％。

Abstract

A mutant Clbeijerinckii strain with higher butanol production ability was obtained by protoplast mutation with 60Co-γ irraditionThe optimal conditions for protoplast preparation were: the cell cultivated for 15 h was dealt with 1 g.L-1 glycin and 0.3 U.L-1 penicillin for 2 h and then digested with 1×104 U.L-1 lysozyme at 37℃ for 8 h.By plate count method

the formation and the regeneration rate of protoplasts was calculated as 99.82% and 10.12% respectively.The protoplasts were then irradiated by 60Co with a dose rate of 5Gy.min-1.The lethality of the protoplasts was 99.3% when the radiation dose was 500 Gy.Under such condition

a mutant strain R3 with higher butanol production ability and higher inheritance stability was selectedCompared with the original strain

the total solvent concentration and the butanol concentration of the mutant strain R3 was increased by 81.46% and 85.41% respectively in the P2 semi-synthesis medium at 37℃.

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Mutation of Cl.beijerinckii protoplasts by 60Co-γ irradiation

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Mutation of Cl.beijerinckii protoplasts by ⁶⁰Co-γ irradiation