| 复合诱变筛选丙烯醇抗性的富马酸高产米根霉菌株 |
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| 引用本文： | 张昆,徐晴,李霜,付永前,高振,黄和.复合诱变筛选丙烯醇抗性的富马酸高产米根霉菌株[J].安徽农业科学,2008,36(23). |
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| 作者姓名： | 张昆 徐晴 李霜 付永前 高振 黄和 |
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| 作者单位： | 南京工业大学制药与生命科学学院,江苏南京,210009;南京工业大学制药与生命科学学院,江苏南京,210009;江苏省工业生物技术创新中心,江苏南京,211816 |
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| 基金项目： | 国家自然科学基金，国家高技术研究发展计划(863计划)，国家重点基础研究发展计划(973计划) |
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| 摘 要： | 目的]筛选具有丙烯醇抗性的富马酸高产米根霉菌株。方法]以溴甲酚绿与丙烯醇平板作为双初筛标准,建立代谢调控选育米根霉富马酸高产菌株的方法。结果]利用紫外诱变对米根霉孢子的最佳诱变条件分别为紫外灯30 W,垂直照射距离30 cm,诱变时间1 min;化学诱变剂亚硝酸浓度0.1 mol/L,诱变时间10 min。优化条件下,通过紫外和化学的复合诱变获得了1株产量高、稳定性好的米根霉丙烯醇抗性突变株ME-F12,富马酸产量达到50.39 g/L,与原始菌株相比提高23%,连续传代5次后,产量无明显降低,具有较高稳定性,副产物乙醇浓度只有1.87 g/L,明显低于原始菌株。结论]为微生物发酵法生产富马酸的工业化奠定了扎实的基础。
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| 关 键 词： | 代谢调控育种 米根霉 富马酸 复合诱变 丙烯醇 |
Integrated Mutagenesis to Scrota High Fumaric Acid Producing Mutant of Rhizopus oryzae with Propylene Alcohol Resistance |
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| ZHANG Kun et al.Integrated Mutagenesis to Scrota High Fumaric Acid Producing Mutant of Rhizopus oryzae with Propylene Alcohol Resistance[J].Journal of Anhui Agricultural Sciences,2008,36(23). |
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| Authors: | ZHANG Kun |
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| Abstract: | Objective]The research aimed to screen high fumaric acid producing mutant of Rhizopus oryzae.Method] Using metabolic engineering method,a two-standard screening approach was devised to select the mutant by bromocresol green and propylene alcohol methods.The enhancement of fumaric acid production from R.oryzae ME-F10 was attempted by ultraviolet(UV) and nitrous acid mutagenesis.Result] After a series screenings,it was found that the optimal condition for UV mutagenesis was a one-minute period,the optimal conditions for nitrous acid mutagenesis were a 10-minute period and nirous acid concentration of 0.1 mol/L.A selected mutant by nitrous acid was subjected to an integrated mutagenesis by UV and nitrous acid.After two steps of inducement,one strain named ME-F12 was selected as an ideal one since it exhibited high fumaric acid production of 50.39 g/L with an increase of 23% than the original strain.It showed high stability with little regression after five generations and it produced little byproduct of 1.87 g/L ethanol,indicating propylene alcohol resistance.Conclusion] The study provided stable foundation for the industrial production of fumaric acid with microbiological fermentation. |
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| Keywords: | Metabolic-engineering directed screening Rhizopus oryzae Fumaric acid Integrated mutagenesis Propylene alcohol |
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