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利用静止细胞培养系统结合提取发酵来研究红曲色素特性和生物合成基因表达的变化。

Variations in Monascus pigment characteristics and biosynthetic gene expression using resting cell culture systems combined with extractive fermentation.

机构信息

School of Biology and Biological Engineering, Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou, 510006, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2018 Jan;102(1):117-126. doi: 10.1007/s00253-017-8576-y. Epub 2017 Nov 2.

Abstract

Monascus pigments are promising sources of natural food colorants, and their productivity can be improved by a novel extractive fermentation technology. In this study, we investigated the variations in pigment characteristics and biosynthetic gene expression levels in resting cell culture systems combined with extractive fermentation in Monascus anka GIM 3.592. Although the biomass was low at about 6 g/L DCW, high pigment titer of approximately 130 AU was obtained in the resting culture with cells from extractive fermentation, illustrating that it had a good biocatalytic activity for pigment synthesis. The oxidation-reduction potential value correlated with the rate of relative content of the intracellular orange pigments to the yellow pigments (O/Y, r > 0.90, p < 0.05), indicating that the change in pigment characteristics may be responsible for the cellular redox activity. The up- or down-regulation of the pigment biosynthetic genes (MpFasA2, MpFasB2, MpPKS5, mppD, mppB, mppR1, and mppR2) in the resting culture with extractive culture cells was demonstrated by real-time quantitative polymerase chain reaction analysis. Moreover, the mppE gene associated with the yellow pigment biosynthesis was significantly (p < 0.05) down-regulated by about 18.6%, whereas the mppC gene corresponding to orange pigment biosynthesis was significantly (p < 0.05) up-regulated by approximately 21.0%. These findings indicated that extractive fermentation was beneficial for the biosynthesis of the intracellular orange pigment. The mechanism described in this study proposes a potential method for the highly efficient production of Monascus pigments.

摘要

红曲色素是很有前途的天然食用色素来源,其生产力可以通过一种新的提取发酵技术来提高。在这项研究中,我们研究了在静止细胞培养系统中结合提取发酵,在 Monascus anka GIM 3.592 中色素特征和生物合成基因表达水平的变化。尽管生物量约为 6 g/L DCW,但从提取发酵的细胞中进行静止培养时,可获得约 130 AU 的高色素滴度,表明其对色素合成具有良好的生物催化活性。氧化还原电位值与细胞内橙色色素与黄色色素的相对含量的比率(O/Y,r > 0.90,p < 0.05)相关,表明色素特征的变化可能是细胞氧化还原活性的原因。通过实时定量聚合酶链反应分析,证明了在提取培养细胞的静止培养中,色素生物合成基因(MpFasA2、MpFasB2、MpPKS5、mppD、mppB、mppR1 和 mppR2)的上调或下调。此外,与黄色色素生物合成相关的 mppE 基因被显著下调(p < 0.05)约 18.6%,而对应橙色色素生物合成的 mppC 基因被显著上调(p < 0.05)约 21.0%。这些发现表明,提取发酵有利于细胞内橙色色素的生物合成。本研究中描述的机制提出了一种高效生产红曲色素的潜在方法。

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