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灵芝液体静置和摇瓶培养中个体灵芝酸的产生和生物合成基因的表达。

Production of individual ganoderic acids and expression of biosynthetic genes in liquid static and shaking cultures of Ganoderma lucidum.

机构信息

State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, China.

出版信息

Appl Microbiol Biotechnol. 2010 Jan;85(4):941-8. doi: 10.1007/s00253-009-2106-5. Epub 2009 Jul 4.

Abstract

Two-stage culture was efficient in enhancing total ganoderic acid (GA) production by Ganoderma lucidum (Fang and Zhong, Biotechnol Prog 18:51-54, 2002). As different GAs have different bioactivities, it is critical to understand the kinetics of individual GA production during fermentation, but no related information is yet available. To understand the regulation of GA biosynthesis, investigation of the accumulation of intermediate (lanosterol) and by-product (ergosterol) and of the expression of three important biosynthetic genes was also conducted in liquid shaking and static cultures of G. lucidum. The results showed that the content of individual GAs increased rapidly in the liquid static culture, and their maximum value was 6- to 25-fold that of shaking culture while lanosterol content in the former was lower than the latter. The transcript of squalene synthase (SQS), lanosterol synthase and 3-hydroxy-3-methylglutaryl coenzyme A reductase in liquid static culture was 4.3-, 2.1-, and 1.9-fold that of the shaking culture, respectively. Higher GA content in liquid static culture was related to increased transcription of those genes especially SQS. The work is helpful to the production of individual GAs and provided an insight into why the liquid static culture was superior to the shaking culture in view of biosynthetic gene expression.

摘要

两段式培养可有效提高灵芝(Fang and Zhong, Biotechnol Prog 18:51-54, 2002)中总灵芝酸(GA)的产量。由于不同的 GA 具有不同的生物活性,因此了解发酵过程中单个 GA 生产的动力学至关重要,但目前尚无相关信息。为了了解 GA 生物合成的调控,还在灵芝的液体摇瓶和静态培养中对中间产物(羊毛甾醇)和副产物(麦角甾醇)的积累以及三个重要生物合成基因的表达进行了研究。结果表明,在液体静态培养中,各个 GA 的含量迅速增加,其最大值是摇瓶培养的 6-25 倍,而前者中的羊毛甾醇含量低于后者。液体静态培养中鲨烯合酶(SQS)、羊毛甾醇合酶和 3-羟-3-甲基戊二酰辅酶 A 还原酶的转录分别是摇瓶培养的 4.3、2.1 和 1.9 倍。液体静态培养中 GA 含量较高与这些基因,尤其是 SQS 的转录增加有关。这项工作有助于各个 GA 的生产,并深入了解为什么从生物合成基因表达的角度来看,液体静态培养优于摇瓶培养。

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