多形汉逊酵母代谢改造生产脂肪酸及发酵条件优化  

Production of fatty acids by engineered Ogataea polymorpha

作　　者：冯叨 高教琪 龚志伟 周雍进[2,3] FENG Dao;GAO Jiaoqi;GONG Zhiwei;ZHOU Yongjin J.(School of Chemistry and Chemical Engineering,Wuhan University of Science and Technology,Wuhan 430080,Hubei,China;Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,Liaoning,China;Dalian Key Laboratory of Energy Biotechnology,Dalian 116023,Liaoning,China)

机构地区：[1]武汉科技大学化学与化工学院,湖北武汉430080 [2]中国科学院大连化学物理研究所生物技术研究部,辽宁大连116023 [3]大连市能源生物技术重点实验室,辽宁大连116023

出　　处：《生物工程学报》2022年第2期760-771,共12页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(21922812,21808216);兴辽英才计划(XLYC1807191);大连市科技创新基金(2019J12GX030)。

摘　　要：脂肪酸作为一种化工原料,在生物能源、化妆品、个人护理产品和工业润滑剂等领域具有广泛应用。多形汉逊酵母因其能够利用甲醇、耐高温、底物谱广等优点,被认为是微生物细胞工厂的理想底盘宿主。本研究首先通过代谢工程构建了产脂肪酸的汉逊酵母细胞工厂。在此基础上,通过发酵条件优化进一步提升了工程菌株生产性能。在温度37℃、pH 6.4、培养基碳氮摩尔比为120、种子液OD_(600)在6–8之间时,摇瓶中工程菌脂肪酸产量达到1.86 g/L。在发酵罐中,采用溶氧(DO)关联法控制补料速度,初始培养基碳氮摩尔比为17.5,在DO高于30%时,补料碳氮摩尔比为120的葡萄糖培养基,脂肪酸产量达到18.0 g/L,显示了汉逊酵母作为脂肪酸合成细胞工厂的潜力,为实现工业化奠定了坚实的理论与应用基础。Fatty acids(FA)are widely used as feed stocks for the production of cosmetics,personal hygiene products,lubricants and biofuels.Ogataea polymorpha is considered as an ideal chassis for bio-manufacturing,due to its outstanding characteristics such as methylotroph,thermal-tolerance and wide substrate spectrum.In this study,we harnessed O.polymorpha for overproduction of fatty acids by engineering its fatty acid metabolism and optimizing the fermentation process.The engineered strain produced 1.86 g/L FAs under the optimized shake-flask conditions(37℃,pH 6.4,a C/N ratio of 120 and an OD_(600) of seed culture of 6–8).The fed-batch fermentation process was further optimized by using a dissolved oxygen(DO)control strategy.The C/N ratio of initial medium was 17.5,and the glucose medium with a C/N ratio of 120 was fed when the DO was higher than 30%.This operation resulted in a titer of 18.0 g/L FA,indicating the potential of using O.polymorpha as an efficient cell factory for the production of FA.

关 键 词：多形汉逊酵母 代谢工程 脂肪酸 限氮发酵 发酵条件优化 

分 类 号：TQ921[轻工技术与工程—发酵工程]