| 代谢工程改造野生耐酸酵母生产L-乳酸 |
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| 引用本文： | 张勤,张梁,丁重阳,王正祥,石贵阳.代谢工程改造野生耐酸酵母生产L-乳酸[J].生物工程学报,2011,27(7):1024-1031. |
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| 作者姓名： | 张勤 张梁 丁重阳 王正祥 石贵阳 |
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| 作者单位： | 江南大学工业生物技术教育部重点实验室，无锡，214122;江南大学生物工程学院生物资源与生物能源研究中心，无锡，214122 |
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| 基金项目： | 中央高校基本科研业务费专项资金 (No. JUSRP31004)，2008年度江苏省高校“青蓝工程”科技创新团队 资助。 |
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| 摘 要： | 以选育低pH条件下高产L-乳酸的酵母菌为目的，从自然样品中筛选分离得到一株能在pH 2.5 (乳酸调节) 的培养基中生长且不利用乳酸的酵母 (初步鉴定为木兰假丝酵母Candida magnolia)；进一步将来源于米根霉As3.819的乳酸脱氢酶编码基因 (ldhA) 插入含有G418抗性基因的酵母穿梭载体，构建了重组质粒pYX212-kanMX-ldhA，电转化入野生型C. magnolia中，筛选获得了一株具有产L-乳酸能力的重组菌株C. magnolia-2；通过发酵实验表明，该重组菌产L-乳酸的最
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| 关 键 词： | 耐酸酵母，米根霉，乳酸脱氢酶，代谢工程，L-乳酸 |
| 收稿时间： | 2010/9/25 0:00:00 |
Metabolic engineering of wild acid-resistant yeast for L-lactic acid production |
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| Qin Zhang,Liang Zhang,Zhongyang Ding,Zhengxiang Wang and Guiyang Shi.Metabolic engineering of wild acid-resistant yeast for L-lactic acid production[J].Chinese Journal of Biotechnology,2011,27(7):1024-1031. |
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| Authors: | Qin Zhang Liang Zhang Zhongyang Ding Zhengxiang Wang and Guiyang Shi |
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| Institution: | Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China |
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| Abstract: | In order to obtain a yeast strain able to produce L-lactic acid under the condition of low pH and high lactate content, one wild acid-resistant yeast strain isolated from natural samples, was found to be able to grow well in YEPD medium (20 g/L glucose, 20 g/L tryptone, 10 g/L yeast extract, adjusted pH 2.5 with lactic acid) without consuming lactic acid. Based on further molecular biological tests, the strain was identified as Candida magnolia. Then, the gene ldhA, encoding a lactate dehydrogenase from Rhizopus oryzae, was cloned into a yeast shuttle vector containing G418 resistance gene. The resultant plasmid pYX212-kanMX-ldhA was introduced into C. magnolia by electroporation method. Subsequently, a recombinant L-lactic acid producing yeast C. magnolia-2 was obtained. The optimum pH of the recombinant yeast is 3.5 for lactic acid production. Moreover, the recombinant strain could grow well and produce lactic acid at pH 2.5. This recombinant yeast strain could be useful for producing L-lactic acid. |
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| Keywords: | acid-resistant yeast Rhizopus oryzae lactate dehydrogenase metabolic engineering L-lactic acid |
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