Zhang H, Kang X, Xiao N, Gao M, Zhao Y, Zhang B, Song Y
Colin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo, Shandong, China.
School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China.
Lett Appl Microbiol. 2019 Mar;68(3):248-257. doi: 10.1111/lam.13111. Epub 2019 Jan 17.
Genetic modification by overexpressing Vitreoscilla haemoglobin (VHb) is an efficient and economic method for improving O utilization for microbial fermentation. In this study, VHb was expressed in oleaginous yeast, Yarrowia lipolytica, and its effect on lipid accumulation was analysed. During fermentation, the dissolved oxygen (DO) concentration was controlled at 5, 10, 20 and 30 of full O saturation and also uncontrolled by varying the stirring speed and aeration rate. Yarrowia lipolytica harbouring the VHb gene (VHb strain) displayed more pseudomycelium than the control strain (VHb strain), and VHb expression also enhanced the cell density. When DO level was controlled at 30%, the biomass of VHb strains reached about 19 g l and increased by 27% compared to VHb strains. Total fatty acid contents, although, were higher in VHb strains than in VHb strains under all DO levels, and maximally increased by c. 40% (from 10·5 to 14·5% of the biomass) when the DO concentration was controlled at 30%. VHb overexpression, however, markedly suppressed citrate secretion. In addition, expression of VHb also induced significant changes in fatty acid composition and increased the oleic acid content. SIGNIFICANCE AND IMPACT OF THE STUDY: Genetic modification by overexpressing Vitreoscilla haemoglobin (VHb) is an efficient and economic method for improving O utilization for microbial fermentation. In this study, VHb was expressed in Yarrowia lipolytica, which is a most attractive model for microbial oil production because of its excellent lipid accumulation capacity and convenient genetic tools. Expression of VHb resulted in lipid accumulation increased and cell morphology shift, however, markedly suppressed citrate secretion. This study provided the new strategy for fermentation technology to improve lipid production in oleaginous micro-organisms.
通过过表达透明颤菌血红蛋白(VHb)进行基因改造是提高微生物发酵中氧气利用效率的一种有效且经济的方法。在本研究中,VHb在产油酵母解脂耶氏酵母中表达,并分析了其对脂质积累的影响。在发酵过程中,通过改变搅拌速度和通气速率,将溶解氧(DO)浓度控制在全氧饱和度的5%、10%、20%和30%,也有未控制的情况。携带VHb基因的解脂耶氏酵母(VHb菌株)比对照菌株(VHb菌株)显示出更多的假菌丝,并且VHb表达也提高了细胞密度。当DO水平控制在30%时,VHb菌株的生物量达到约19 g/L,比VHb菌株增加了27%。然而,在所有DO水平下,VHb菌株中的总脂肪酸含量均高于VHb菌株,当DO浓度控制在30%时,总脂肪酸含量最大增加约40%(从生物量的10.5%增加到14.5%)。然而,VHb过表达显著抑制了柠檬酸盐的分泌。此外,VHb的表达还引起了脂肪酸组成的显著变化,并增加了油酸含量。研究的意义和影响:通过过表达透明颤菌血红蛋白(VHb)进行基因改造是提高微生物发酵中氧气利用效率的一种有效且经济的方法。在本研究中,VHb在解脂耶氏酵母中表达,由于其出色的脂质积累能力和便捷的遗传工具,解脂耶氏酵母是微生物油脂生产中最具吸引力的模型。VHb的表达导致脂质积累增加和细胞形态改变,然而,显著抑制了柠檬酸盐的分泌。本研究为发酵技术提高产油微生物的脂质产量提供了新策略。