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通过在灵芝中组成型表达透明颤菌血红蛋白基因提高灵芝酸产量

Enhancement of ganoderic acid production by constitutively expressing Vitreoscilla hemoglobin gene in Ganoderma lucidum.

作者信息

Li Huan-Jun, He Yi-Long, Zhang De-Huai, Yue Tong-Hui, Jiang Lu-Xi, Li Na, Xu Jun-Wei

机构信息

Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.

Faculty of Science, Kunming University of Science and Technology, Kunming, 6500500, China.

出版信息

J Biotechnol. 2016 Jun 10;227:35-40. doi: 10.1016/j.jbiotec.2016.04.017. Epub 2016 Apr 11.

Abstract

The Vitreoscilla hemoglobin (VHb) gene was expressed in Ganoderma lucidum to enhance antitumor ganoderic acid (GA) production. The effects of VHb expression on the accumulation of GAs and lanosterol (intermediate) and the transcription of GA biosynthesis genes were also investigated. In VHb-expressing G. lucidum, the maximum concentrations of four individual GAs (GA-S, GA-T, GA-Mk and GA-Me) were 19.1±1.8, 34.6±2.1, 191.5±13.1 and 45.2±2.8μg/100mg dry weight, respectively, which were 1.4-, 2.2, 1.9- and 2.0-fold higher than those obtained in the wild-type strain. Moreover, the maximum lanosterol concentration in the strain expressing VHb was 1.28-fold lower than that in the wild-type strain. The transcription levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase, squalene synthase, and lanosterol synthase genes were up-regulated by 1.6-, 1.5-, and 1.6-fold, respectively, in the strain expressing VHb. This work is beneficial in developing an efficient fermentation process for the hyperproduction of GAs.

摘要

将透明颤菌血红蛋白(VHb)基因在灵芝中表达,以提高抗肿瘤灵芝酸(GA)的产量。还研究了VHb表达对灵芝酸和羊毛甾醇(中间体)积累以及GA生物合成基因转录的影响。在表达VHb的灵芝中,四种单一灵芝酸(GA-S、GA-T、GA-Mk和GA-Me)的最大浓度分别为19.1±1.8、34.6±2.1、191.5±13.1和45.2±2.8μg/100mg干重,分别比野生型菌株高出1.4倍、2.2倍、1.9倍和2.0倍。此外,表达VHb的菌株中羊毛甾醇的最大浓度比野生型菌株低1.28倍。在表达VHb的菌株中,3-羟基-3-甲基戊二酰辅酶A还原酶、角鲨烯合酶和羊毛甾醇合酶基因的转录水平分别上调了1.6倍、1.5倍和1.6倍。这项工作有助于开发一种高效发酵工艺,用于灵芝酸的超量生产。

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