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首页> 外文期刊>Applied Microbiology and Biotechnology >Strain improvement of Sporolactobacillus inulinus ATCC 15538 for acid tolerance and production of D-lactic acid by genome shuffling
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Strain improvement of Sporolactobacillus inulinus ATCC 15538 for acid tolerance and production of D-lactic acid by genome shuffling

机译：菊粉菌ATCC 15538的耐酸菌株改良和基因组改组生产D-乳酸

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摘要

Improvement of acid tolerance and production of D-lactic acid by Sporolactobacillus inulinus ATCC 15538 was performed by using recursive protoplast fusion in a genome shuffling format. The starting population was generated by ultraviolet irradiation, diethyl sulfate mutagenesis, and pH-gradient filter and then, subjected for the recursive protoplast fusion. The concentration of lysozyme, time, and temperature for enzyme treatment were optimized by response surface methodology based on the central composite design. Based on contour plots and variance analysis, the model predicted a maximum Y (multiply protoplasts formation ratio by protoplasts regeneration ratio), 60.4%, and the corresponding above used values were 7.75 mg/ml lysozyme, 1.59 h, and 38°C. A pH-5-resistant recombinant, F3-4, was obtained after three rounds of genome shuffling and its production of D-lactic acid reached 93.4 g/l in a 5 L bioreactor, which was increased by 39.8% and 119% in comparison with that of UV generated strain and the original strain S. inulinus ATCC 15538, respectively. The subculture experiments indicated that F3-4 was genetically stable. Keywords D-lactic acid - Sporolactobacillus inulinus ATCC 15538 - Genome shuffling - Protoplast fusion - Response surface methodology (RSM) - Breeding
机译：通过使用基因组改组格式的递归原生质体融合来提高菊粉孢菌ATCC 15538的耐酸性和D-乳酸的产生。通过紫外线照射，硫酸二乙酯诱变和pH梯度过滤器生成起始种群，然后进行递归原生质体融合。基于中心复合设计，通过响应面法优化了酶处理的溶菌酶浓度，时间和温度。基于轮廓图和方差分析，该模型预测最大Y（乘以原生质体再生率乘以原生质体形成率）为60.4％，相应的上述使用值为7.75 mg / ml溶菌酶，1.59 h和38°C。经过三轮基因组改组后，获得了耐pH-5的重组体F3-4，在5 L生物反应器中D-乳酸的产量达到93.4 g / l，与之相比分别增加了39.8％和119％用紫外线产生的菌株和最初的菌株菊粉链球菌ATCC 15538。继代培养实验表明F3-4是遗传稳定的。关键词D-乳酸-菊粉孢菌ATCC 15538-基因组改组-原生质体融合-响应面法（RSM）-育种

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