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红海藻类树状软骨藻中环阿屯醇合酶的克隆与功能表征

Cloning and Functional Characterization of Cycloartenol Synthase from the Red Seaweed Laurencia dendroidea.

作者信息

Calegario Gabriela, Pollier Jacob, Arendt Philipp, de Oliveira Louisi Souza, Thompson Cristiane, Soares Angélica Ribeiro, Pereira Renato Crespo, Goossens Alain, Thompson Fabiano L

机构信息

Departament of Marine Biology, Federal Fluminense University (UFF), Niterói, Brazil.

Institute of Biology, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.

出版信息

PLoS One. 2016 Nov 10;11(11):e0165954. doi: 10.1371/journal.pone.0165954. eCollection 2016.

Abstract

The red seaweed Laurencia dendroidea belongs to the Rhodophyta, a phylum of eukaryotic algae that is widely distributed across the oceans and that constitute an important source of bioactive specialized metabolites. Laurencia species have been studied since 1950 and were found to contain a plethora of specialized metabolites, mainly halogenated sesquiterpenes, diterpenes and triterpenes that possess a broad spectrum of pharmacological and ecological activities. The first committed step in the biosynthesis of triterpenes is the cyclization of 2,3-oxidosqualene, an enzymatic reaction carried out by oxidosqualene cyclases (OSCs), giving rise to a broad range of different compounds, such as the sterol precursors cycloartenol and lanosterol, or triterpene precursors such as cucurbitadienol and β-amyrin. Here, we cloned and characterized the first OSC from a red seaweed. The OSC gene was identified through mining of a L. dendroidea transcriptome dataset and subsequently cloned and heterologously expressed in yeast for functional characterization, which indicated that the corresponding enzyme cyclizes 2,3-oxidosqualene to the sterol precursor cycloartenol. Accordingly, the gene was named L. dendroidea cycloartenol synthase (LdCAS). A phylogenetic analysis using OSCs genes from plants, fungi and algae revealed that LdCAS grouped together with OSCs from other red algae, suggesting that cycloartenol could be the common product of the OSC in red seaweeds. Furthermore, profiling of L. dendroidea revealed cholesterol as the major sterol accumulating in this species, implicating red seaweeds contain a 'hybrid' sterol synthesis pathway in which the phytosterol precursor cycloartenol is converted into the major animal sterol cholesterol.

摘要

红藻树状软骨藻属于红藻门,红藻门是一类真核藻类,广泛分布于各大洋,是生物活性特殊代谢产物的重要来源。自1950年以来,人们一直在研究软骨藻属物种,发现它们含有大量特殊代谢产物,主要是具有广泛药理和生态活性的卤化倍半萜、二萜和三萜。三萜生物合成的第一个关键步骤是2,3-氧化角鲨烯的环化,这是一种由氧化角鲨烯环化酶(OSC)催化的酶促反应,会产生多种不同的化合物,如甾醇前体环阿屯醇和羊毛甾醇,或三萜前体如葫芦二烯醇和β-香树脂醇。在此,我们克隆并鉴定了首个来自红藻的OSC。通过挖掘树状软骨藻转录组数据集鉴定出该OSC基因,随后将其克隆并在酵母中进行异源表达以进行功能鉴定,结果表明相应的酶将2,3-氧化角鲨烯环化为甾醇前体环阿屯醇。因此,该基因被命名为树状软骨藻环阿屯醇合酶(LdCAS)。使用来自植物、真菌和藻类的OSC基因进行的系统发育分析表明,LdCAS与其他红藻的OSC聚在一起,这表明环阿屯醇可能是红藻中OSC的常见产物。此外,对树状软骨藻的分析表明,胆固醇是该物种中积累的主要甾醇,这意味着红藻含有一种“混合”甾醇合成途径,其中植物甾醇前体环阿屯醇被转化为主要的动物甾醇胆固醇。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/232b/5104453/1c6597fc1217/pone.0165954.g001.jpg

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