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油脂真菌 WJ11 中二酰基甘油酰基转移酶的同源过表达增强了静态固体培养下的油脂积累。

Homologous Overexpression of Diacylglycerol Acyltransferase in Oleaginous Fungus WJ11 Enhances Lipid Accumulation under Static Solid Cultivation.

机构信息

Colin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo, Shandong 255000, People's Republic of China.

出版信息

J Agric Food Chem. 2022 Jul 27;70(29):9073-9083. doi: 10.1021/acs.jafc.2c03489. Epub 2022 Jul 18.

Abstract

Diacylglycerol acyltransferase (DGAT) catalyzes the binding of acyl-CoA to diacylglycerol to form triacylglycerol (TAG). Previous studies strongly indicate that DGAT2, rather than DGAT1, is crucial for TAG accumulation in the oleaginous fungus . To increase the lipid content of WJ11, McDGAT2 was overexpressed by homologous recombination; compared to the control strain Mc2075, transformants McDGAT2d showed a significant increase in biomass for both spores and mycelia (from 87.7 to 101.2 mg/g in spores and from 75.6 to 93.1 mg/g in mycelia). McDGAT2 overexpression under static solid fermentation gave a greater boost to lipid accumulation in mycelia than in spores. Total fatty acid content in mycelia increased by 68.0% (from 13.6 to 22.8%) and in spores by 26.3% (from 10.6 to 13.4%). However, under submerged fermentation, the lipid content of McDGAT2d was the same as the control, while biomass was slightly reduced. Transcriptomics showed that NADPH was derived mainly from the pentose phosphate pathway, acetyl-CoA was from multiple pathways, and leucine metabolism played an important role in substrate supply for fatty acid biosynthesis. Static solid fermentation may be the more suitable fermentation method for microbial oil production by filamentous fungi due to its lower fermentation costs.

摘要

二酰基甘油酰基转移酶 (DGAT) 催化酰基辅酶 A 与二酰基甘油结合形成三酰基甘油 (TAG)。先前的研究强烈表明,DGAT2 而不是 DGAT1,对于产油真菌中 TAG 的积累至关重要。为了提高 WJ11 的油脂含量,通过同源重组过表达了 McDGAT2;与对照菌株 Mc2075 相比,转化子 McDGAT2d 的孢子和菌丝体的生物量都显著增加(从 87.7 毫克/克增加到 101.2 毫克/克,从 75.6 毫克/克增加到 93.1 毫克/克)。在静态固体发酵下,McDGAT2 的过表达对菌丝体中的油脂积累的促进作用大于对孢子的促进作用。菌丝体中的总脂肪酸含量增加了 68.0%(从 13.6 毫克/克增加到 22.8 毫克/克),而孢子中的总脂肪酸含量增加了 26.3%(从 10.6 毫克/克增加到 13.4 毫克/克)。然而,在液体发酵下,McDGAT2d 的油脂含量与对照相同,而生物量略有下降。转录组学表明,NADPH 主要来自戊糖磷酸途径,乙酰辅酶 A 来自多个途径,亮氨酸代谢在脂肪酸生物合成的底物供应中起着重要作用。由于其较低的发酵成本,静态固体发酵可能是丝状真菌微生物油脂生产更合适的发酵方法。

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