Metabolic engineering Corynebacterium glutamicum for D-chiro-inositol production

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Abstract

D-chiro-inositol (DCI) is a potential drug for the treatment of type II diabetes and polycystic ovary syndrome. In order to effectively synthesize DCI in Corynebacterium glutamicum, the genes related to inositol catabolism in clusters iol1 and iol2 were knocked out in C. glutamicum SN01 to generate the chassis strain DCI-1. DCI-1 did not grow in and catabolize myo-inositol (MI). Subsequently, different exogenous and endogenous inosose isomerases were expressed in DCI-1 and their conversion ability of DCI from MI were compared. After fermentation, the strain DCI-7 co-expressing inosose isomerase IolI2 and inositol dehydrogenase IolG was identified as the optimal strain. Its DCI titer reached 3.21 g/L in the presence of 20 g/L MI. On this basis, the pH, temperature and MI concentration during whole-cell conversion of DCI by strain DCI-7 were optimized. Finally, the optimal condition that achieved the highest DCI titer of 6.96 g/L were obtained at pH 8.0, 37 °C and addition of 40 g/L MI. To our knowledge, it is the highest DCI titer ever reported.

谷氨酸棒状杆菌生产 D-螺肌醇的代谢工程
摘要 D-螺肌醇(DCI)是治疗II型糖尿病和多囊卵巢综合征的潜在药物。为了在谷氨酸棒状杆菌(Corynebacterium glutamicum)中有效合成 DCI,我们敲除了谷氨酸棒状杆菌 SN01 中 iol1 和 iol2 群组中与肌醇分解代谢相关的基因,生成了基质菌株 DCI-1。DCI-1 不在肌醇(MI)中生长,也不分解肌醇(MI)。随后,在 DCI-1 中表达了不同的外源性和内源性肌糖异构酶,并比较了它们从 MI 转化 DCI 的能力。经过发酵,共同表达肌醇异构酶 IolI2 和肌醇脱氢酶 IolG 的菌株 DCI-7 被确定为最佳菌株。在 20 克/升 MI 的条件下,其 DCI 滴度达到 3.21 克/升。在此基础上,对菌株 DCI-7 全细胞转化 DCI 过程中的 pH 值、温度和 MI 浓度进行了优化。最后,在 pH 值为 8.0、温度为 37 °C、添加 40 克/升 MI 的条件下,DCI 滴度最高,达到 6.96 克/升。据我们所知,这是迄今为止报道的最高 DCI 滴度。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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