丁二酸高产菌株的选育及发酵条件优化

doi:10.3969/j.issn.1000-1565.2018.02.010

摘要/Abstract

摘要： 对产琥珀酸放线杆菌RH进行紫外诱变选育,用TLC法初筛得到84株丁二酸产量较高的菌株;对初筛得到的菌株进一步用HPLC法复筛,得到1株高产突变菌株C4-1.经遗传稳定性分析,发现该高产突变株连续传5代后丁二酸产量基本保持稳定.碳源、氮源优化实验表明,突变株C4-1产丁二酸的最适碳源为液化淀粉,最适氮源为玉米浆干粉,且可以采用液化玉米粉添加少量玉米浆干粉代替液化淀粉和玉米浆干粉,以节约成本;在优化条件下突变株C4-1三角瓶发酵60 h,丁二酸产量达到(52.91±0.53)g/L,较原始菌株提高了33.46%.

关键词: 产琥珀酸放线杆菌, 丁二酸, 高产突变株, 紫外诱变, 发酵优化

Abstract: The method of UV mutagenic breeding was carried out for Actinobacillus succinogenes RH to screen succinic acid high-yielding strain.84 mutant strains with relative high succinic acid production were obtained in preliminary screening by TLC method,and a high-yielding mutant strain C4-1 was got by further screening using HPLC.It was found that strain C4-1 remained stable succinic acid producing ability after continuous passage for five generations through the genetic stability analysis.Optimization experiments of carbon source and nitrogen source showed that the optimal carbon source and nitrogen source for succinic acid fermentation were liquefied corn starch(LCS)and corn steep liquor(CSL),respectively.Further experiment showed that the carbon source and nitrogen source can be replaced by liquefied corn flour(LCF)and small amount of CSL so as to reduce the cost.Taking the advantage of optimal conditions,the mutant strain C4-1 resulted in the succinic acid yield of(52.91±0.53)g/L after fermented for- DOI:10.3969/j.issn.1000-1565.2018.02.010丁二酸高产菌株的选育及发酵条件优化苏晓娆¹,郑延红¹,席丹¹,吕志堂^1,2(1.河北大学生命科学学院,河北省微生物多样性研究与应用重点实验室,河北保定 071002;2.河北大学药物化学与分子诊断教育部重点实验室,河北保定 071002)摘要:对产琥珀酸放线杆菌RH进行紫外诱变选育,用TLC法初筛得到84株丁二酸产量较高的菌株;对初筛得到的菌株进一步用HPLC法复筛,得到1株高产突变菌株C4-1.经遗传稳定性分析,发现该高产突变株连续传5代后丁二酸产量基本保持稳定.碳源、氮源优化实验表明,突变株C4-1产丁二酸的最适碳源为液化淀粉,最适氮源为玉米浆干粉,且可以采用液化玉米粉添加少量玉米浆干粉代替液化淀粉和玉米浆干粉,以节约成本;在优化条件下突变株C4-1三角瓶发酵60 h,丁二酸产量达到(52.91±0.53)g/L,较原始菌株提高了33.46%.关键词:产琥珀酸放线杆菌;丁二酸;高产突变株;紫外诱变;发酵优化中图分类号:Q939.9 文献标志码:A 文章编号:1000-1565(2018)02-0177-08Breeding and fermentation conditions optimization of a high-yielding succinic acid producing strainSU Xiaorao¹,ZHENG Yanhong¹,XI Dan¹,LÜ Zhitang^1,2(1.Key Laboratory of Microbial Diversity Research and Application of Hebei Province,College of Life Sciences,Hebei University,Baoding 071002,China;2.Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education,Hebei University,Baoding 071002,China)Abstract:The method of UV mutagenic breeding was carried out for Actinobacillus succinogenes RH to screen succinic acid high-yielding strain.84 mutant strains with relative high succinic acid production were obtained in preliminary screening by TLC method,and a high-yielding mutant strain C4-1 was got by further screening using HPLC.It was found that strain C4-1 remained stable succinic acid producing ability after continuous passage for five generations through the genetic stability analysis.Optimization experiments of carbon source and nitrogen source showed that the optimal carbon source and nitrogen source for succinic acid fermentation were liquefied corn starch(LCS)and corn steep liquor(CSL),respectively.Further experiment showed that the carbon source and nitrogen source can be replaced by liquefied corn flour(LCF)and small amount of CSL so as to reduce the cost.Taking the advantage of optimal conditions,the mutant strain C4-1 resulted in the succinic acid yield of(52.91±0.53)g/L after fermented for- 收稿日期:2017-11-22 基金项目:河北大学自然科学研究计划项目(799207217072) 第一作者:苏晓娆(1991—),女,河北石家庄人,河北大学在读硕士研究生,主要从事微生物遗传育种研究.E-mail:465738724@qq.com 通信作者:吕志堂(1973—),男,河北曲周人,河北大学教授,博士,主要从事微生物资源与系统学研究.E-mail:lzt325@126.com第2期苏晓娆等:丁二酸高产菌株的选育及发酵条件优化60 h in flask with 33.46% high that the original strain.

Key words: Actinobacillus succinogenes, succinic acid, high-yielding mutant strain, ultraviolet mutagenesis, fermentation optimization

中图分类号:

Q939.9

苏晓娆,郑延红,席丹,吕志堂. 丁二酸高产菌株的选育及发酵条件优化[J]. 河北大学学报(自然科学版), 2018, 38(2): 177-184.

SU Xiaorao,ZHENG Yanhong,XI Dan,LV Zhitang. Breeding and fermentation conditions optimization of a high-yielding succinic acid producing strain[J]. Journal of Hebei University (Natural Science Edition), 2018, 38(2): 177-184.

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