长链酰基辅酶 A 合成酶基因家族的功能特征从油脂藻类。

Functional Characterization of Long-Chain Acyl-CoA Synthetase Gene Family from the Oleaginous Alga .

机构信息

Laboratory for Algae Biotechnology & Innovation, College of Engineering, Peking University, Beijing 100871, China.

BIC-ESAT, College of Engineering, Peking University, Beijing 100871, China.

出版信息

J Agric Food Chem. 2020 Apr 15;68(15):4473-4484. doi: 10.1021/acs.jafc.0c01284. Epub 2020 Apr 6.

DOI:10.1021/acs.jafc.0c01284

PMID:32208653

Abstract

Long-chain acyl-coenzyme A (CoA) synthetase (LACS) catalyzes the formation of acyl-CoAs from free fatty acids, which is pivotal for lipid metabolism. Here, we confirmed the presence of six genes in . Functional complementation and in vitro enzymatic assay indicated that CzLACS2 through CzLACS5 rather than CzLACS1 or CzLACS6 are bona fide LACS enzymes and they have overlapping yet distinct substrate preference. The results of the subcellular colocalization experiment and different expression patterns under three triacylglycerol (TAG)-inducing conditions showed that CzLACS2 through CzLACS4 reside at endoplasmic reticulum (ER) and are involved in TAG biosynthesis, while CzLACS5 resides in peroxisome and participates in fatty acid β-oxidation. The yeast one-hybrid assay using a library of 50 transcription factors (TFs) constructed in our study identified 12 TFs potentially involved in regulating the expression of s. Moreover, heterologous expression of s demonstrated their engineering potential for modulating TAG synthesis in yeast and algal cells.

摘要

长链酰基辅酶 A（CoA）合成酶（LACS）催化游离脂肪酸形成酰基辅酶 A，这对于脂质代谢至关重要。在这里，我们在中证实了 6 个基因的存在。功能互补和体外酶活性测定表明，CzLACS2 到 CzLACS5 而不是 CzLACS1 或 CzLACS6 是真正的 LACS 酶，它们具有重叠但不同的底物偏好。亚细胞共定位实验和三种三酰基甘油（TAG）诱导条件下的不同表达模式的结果表明，CzLACS2 到 CzLACS4 位于内质网（ER）中并参与 TAG 生物合成，而 CzLACS5 位于过氧化物酶体中并参与脂肪酸β-氧化。使用我们研究中构建的 50 个转录因子（TF）文库进行的酵母单杂交测定鉴定出 12 个可能参与调节 s 表达的 TF。此外，s 的异源表达证明了它们在酵母和藻类细胞中调节 TAG 合成的工程潜力。