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米根霉的替代呼吸和富马酸生成。

Alternative respiration and fumaric acid production of Rhizopus oryzae.

机构信息

State Key Laboratory of Material-oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, No. 5 Xinmofan Road, Nanjing, 210009, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2014 Jun;98(11):5145-52. doi: 10.1007/s00253-014-5615-9. Epub 2014 Mar 19.

Abstract

Under the conditions of fumaric acid fermentation, Rhizopus oryzae ME-F14 possessed at least two respiratory systems. The respiration of mycelia was partially inhibited by the cytochrome respiration inhibitor antimycin A or the alternative respiration inhibitor salicylhydroxamic acid and was completely inhibited in the presence of both antimycin A and salicylhydroxamic acid. During fumaric acid fermentation process, the activity of alternative respiration had a great correlation with fumaric acid productivity; both of them reached peak at the same time. The alternative oxidase gene, which encoded the mitochondrial alternative oxidase responsible for alternative respiration in R. oryzae ME-F14, was cloned and characterized in Escherichia coli. The activity of alternative respiration, the alternative oxidase gene transcription level, as well as the fumaric acid titer were measured under different carbon sources and different carbon-nitrogen ratios. The activity of alternative respiration was found to be comparable to the transcription level of the alternative oxidase gene and the fumaric acid titer. These results indicated that the activity of the alternative oxidase was regulated at the transcription stage under the conditions tested for R. oryzae ME-F14.

摘要

在富马酸发酵条件下,米根霉 ME-F14 至少具有两种呼吸系统。细胞色素呼吸抑制剂安密妥 A 或替代呼吸抑制剂水杨羟肟酸部分抑制菌丝的呼吸,而安密妥 A 和水杨羟肟酸同时存在时则完全抑制呼吸。在富马酸发酵过程中,替代呼吸活性与富马酸生产力有很大的相关性;它们都在同一时间达到峰值。在大肠杆菌中克隆并表征了编码米根霉 ME-F14 中负责替代呼吸的线粒体替代氧化酶的替代氧化酶基因。在不同碳源和不同碳氮比下测量了替代呼吸活性、替代氧化酶基因转录水平以及富马酸效价。发现替代呼吸活性与替代氧化酶基因的转录水平和富马酸效价相当。这些结果表明,在所测试的米根霉 ME-F14 条件下,替代氧化酶的活性在转录阶段受到调节。

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