章鱼萜环化酶的发现和表征方法。

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology
Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-03-05 DOI:10.1016/bs.mie.2024.02.011
Immo Burkhardt, Lara Dürr, Natalie E Grayson, Bradley S Moore
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引用次数: 0

摘要

章鱼是海洋环境中萜类化合物最丰富的来源,迄今为止,该门类已知有 4000 多种不同的化合物。然而,这些化合物的生物化学和遗传学来源一直难以确定,直到最近的研究表明,八珊瑚自身的染色体 DNA 中编码了负责萜类化合物生物合成的基因,而不是像最初提出的那样来自微生物共生体。已确定的珊瑚基因包括编码一类新的萜烯环化酶(TCs)的基因,这些基因与其他候选的剪裁酶类聚集在一起。系统发生学分析表明,章鱼萜环化酶是一个单系支系,有别于植物、细菌和其他生物的萜环化酶。新发现的TCs群似乎在章鱼中无处不在,而且在进化上是古老的。鉴于八瓣珊瑚萜类生物化学的发现时间不长,而且目前只有有限的基因组数据,因此从八瓣珊瑚中发现新的萜类生产生物合成途径具有很大的潜力。下一章概述了章鱼 DNA 和 RNA 提取、基因组和转录组组装和挖掘、TC 克隆和基因表达、蛋白质纯化和体外分析的实用实验程序。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Methods for the discovery and characterization of octocoral terpene cyclases.

Octocorals are the most prolific source of terpenoids in the marine environment, with more than 4000 different compounds known from the phylum to date. However, the biochemical and genetic origin of their production remained elusive until recent studies showed that octocorals encode genes responsible for the biosynthesis of terpenoids in their own chromosomal DNA rather than from microbial symbionts as originally proposed. The identified coral genes include those encoding a new group of class I terpene cyclases (TCs) clustered among other candidate classes of tailoring enzymes. Phylogenetic analyses established octocoral TCs as a monophyletic clade, distinct from TCs of plants, bacteria, and other organisms. The newly discovered group of TCs appears to be ubiquitous in octocorals and is evolutionarily ancient. Given the recent discovery of octocoral terpenoid biochemistry and only limited genomic data presently available, there is substantial potential for discovering new biosynthetic pathways from octocorals for terpene production. The following chapter outlines practical experimental procedures for octocoral DNA and RNA extraction, genome and transcriptome assembly and mining, TC cloning and gene expression, protein purification, and in vitro analyses.

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来源期刊
Methods in enzymology
Methods in enzymology 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
308
审稿时长
3-6 weeks
期刊介绍: The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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