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裂殖壶菌中脂肪酸的产生:多不饱和脂肪酸合酶和I型脂肪酸合酶的作用

Fatty acid production in Schizochytrium sp.: Involvement of a polyunsaturated fatty acid synthase and a type I fatty acid synthase.

作者信息

Hauvermale A, Kuner J, Rosenzweig B, Guerra D, Diltz S, Metz J G

机构信息

Martek Biosciences Boulder Corporation, Boulder, CO 80301, USA.

出版信息

Lipids. 2006 Aug;41(8):739-47. doi: 10.1007/s11745-006-5025-6.

Abstract

Schizochytrium sp. is a marine microalga that has been developed as a commercial source for docosahexaenoic acid (DHA, C22:6 (omega-3), enriched biomass, and oil. Previous work suggested that the DHA, as well as docosapentaenoic acid (DPA, C22:5 omega-6), that accumulate in Schizochytrium are products of a multi-subunit polyunsaturated fatty acid (PUFA) synthase (1). Here we show data to support this view and also provide information on other aspects of fatty acid synthesis in this organism. Three genes encoding subunits of the PUFA synthase were isolated from genomic DNA and expressed in E. coli along with an essential accessory gene encoding a phosphopantetheinyl transferase (PPTase). The resulting transformants accumulated both DHA and DPA. The ratio of DHA to DPA was approximately the same as that observed in Schizochytrium. Treatment of Schizochytrium cells with certain levels of cerulenin resulted in inhibition of 14C acetate incorporation into short chain fatty acids without affecting labeling of PUFAs, indicating distinct biosynthetic pathways. A single large gene encoding the presumed short chain fatty acid synthase (FAS) was cloned and sequenced. Based on sequence homology and domain organization, the Schizochytrium FAS resembles a fusion of fungal FAS beta and alpha subunits.

摘要

裂殖壶菌属是一种海洋微藻,已被开发成为二十二碳六烯酸(DHA,C22:6(ω-3))、富含DHA的生物质以及油脂的商业来源。先前的研究表明,裂殖壶菌中积累的DHA以及二十二碳五烯酸(DPA,C22:5 ω-6)是一种多亚基多不饱和脂肪酸(PUFA)合酶的产物(1)。在此,我们展示数据支持这一观点,并提供该生物体脂肪酸合成其他方面的信息。从基因组DNA中分离出编码PUFA合酶亚基的三个基因,并与一个编码磷酸泛酰巯基乙胺基转移酶(PPTase)的必需辅助基因一起在大肠杆菌中表达。所得转化体积累了DHA和DPA。DHA与DPA的比例与在裂殖壶菌中观察到的大致相同。用一定水平的浅蓝菌素处理裂殖壶菌细胞,导致14C乙酸掺入短链脂肪酸受到抑制,而不影响多不饱和脂肪酸的标记,这表明存在不同的生物合成途径。克隆并测序了一个编码假定的短链脂肪酸合酶(FAS)的单一大型基因。基于序列同源性和结构域组织,裂殖壶菌FAS类似于真菌FASβ和α亚基的融合体。

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