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通过增强乙酰辅酶 A 和 ATP 的供应来实现 Aureobasidium melanogenum 9-1 的代谢工程改造,以实现 Liamocins 的过量生产。

Metabolic engineering of Aureobasidium melanogenum 9-1 for overproduction of liamocins by enhancing supply of acetyl-CoA and ATP.

机构信息

College of Marine Life Sciences,Ocean University of China, Yushan Road, No. 5, Qingdao, China.

College of Marine Life Sciences,Ocean University of China, Yushan Road, No. 5, Qingdao, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, 266003, China.

出版信息

Microbiol Res. 2022 Dec;265:127172. doi: 10.1016/j.micres.2022.127172. Epub 2022 Aug 28.

Abstract

In this study, it was found that reducing consumption of acetyl-CoA in mitochondria, peroxisome and lipid biosynthesis could not obviously enhance liamocin biosynthesis by engineered strains of Aureobasidium melanogenm 9-1, but decreased cell growth of the mutants. On the contrary, expression of heterologous PTA gene for phosphotransacetylase in PK pathway and native ALD gene for acetaldehyde dehydrogenase and ACS gene encoding acetyl-CoA synthetase in the PDH bypass pathway reduced liamocin biosynthesis. However, expression the PK gene for phosphoketolase, the PDC gene encoding pyruvate decarboxylase and VHb gene coding for Vitreoscilla hemoglobin (VHb) in the glucose derepression mutants could greatly enhance liamocin production. The resulting strain V33 could produce 55.38 g/L of liamocin and 25.10 g/L of cell dry weight from 117.27 g/L of glucose within 168 h of 10-liter fermentation, leading to the yield of 0.47 g/g of glucose, the productivity of 0.33 g/L/h and rate of glucose utilization of 0.70 ± 0.01 g/L/h. This was a new and efficient strategy for overproduction of liamocin by A. melanogenm.

摘要

在这项研究中,发现减少线粒体、过氧化物酶体和脂类生物合成中的乙酰辅酶 A 的消耗,不能明显增强米黑根毛霉 9-1 工程菌株的利亚霉素生物合成,但降低了突变体的细胞生长。相反,在 PDH 旁路途径中表达异源 PTA 基因(磷酸转乙酰酶)、天然 ALD 基因(乙醛脱氢酶)和 ACS 基因(乙酰辅酶 A 合成酶),减少了利亚霉素的生物合成。然而,在葡萄糖去阻遏突变体中表达 PK 基因(磷酸酮醇酶)、PDC 基因(丙酮酸脱羧酶)和 VHb 基因(编码玻璃球菌血红蛋白,VHb),可以大大增强利亚霉素的生产。所得菌株 V33 可以在 10 升发酵 168 小时内,从 117.27 克/升葡萄糖中产生 55.38 克/升利亚霉素和 25.10 克/升细胞干重,导致 0.47 克/克葡萄糖的产率、0.33 克/升/小时的生产率和 0.70±0.01 克/升/小时的葡萄糖利用率。这是一种通过米黑根毛霉过量生产利亚霉素的新的有效策略。

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