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来自粘细菌黄色粘球菌的去饱和酶的功能分析。

Functional analysis of desaturases from the myxobacterium Myxococcus xanthus.

作者信息

Ring Michael W, Bode Edna, Schwär Gertrud, Bode Helge B

机构信息

Department of Pharmaceutical Biotechnology, Saarland University, Saarbrücken, Germany.

出版信息

FEMS Microbiol Lett. 2009 Jul;296(1):124-30. doi: 10.1111/j.1574-6968.2009.01634.x. Epub 2009 Apr 28.

Abstract

The fatty acid (FA) profiles of myxobacteria contain FA species with double bonds at the Delta(5) and Delta(11) positions, the latter being rather unusual among bacteria. Despite this knowledge, the mechanism for introduction of these double bonds has never been described before in myxobacteria. Searches for candidate genes in the genome of the model organism Myxococcus xanthus revealed 16 genes, which have been annotated as FA desaturases. However, due to redundant substrate specificity, functional analyses of these enzymes by construction of inactivation mutants did not lead to the identification of their function or substrate specificity. Therefore, we elucidated the regioselectivity of the desaturation reactions by heterologous expression of eight desaturases from M. xanthus in Pseudomonas putida and thus could prove five of them to be indeed active as desaturases, with three (MXAN_1742, MXAN_3495 and MXAN_5461) and two (MXAN_0317 and MXAN_6306) acting as Delta(5) and Delta(11) desaturases, respectively. This is the first report about the heterologous expression and regioselectivity of FA desaturases in myxobacteria.

摘要

粘细菌的脂肪酸(FA)谱含有在Δ(5)和Δ(11)位置带有双键的脂肪酸种类,后者在细菌中相当罕见。尽管有这一认识,但在粘细菌中,引入这些双键的机制此前从未被描述过。在模式生物黄色粘球菌的基因组中搜索候选基因,发现了16个被注释为FA去饱和酶的基因。然而,由于底物特异性冗余,通过构建失活突变体对这些酶进行功能分析,并未确定它们的功能或底物特异性。因此,我们通过在恶臭假单胞菌中异源表达来自黄色粘球菌的8种去饱和酶,阐明了去饱和反应的区域选择性,从而证明其中5种确实具有去饱和酶活性,其中3种(MXAN_1742、MXAN_3495和MXAN_5461)和2种(MXAN_0317和MXAN_6306)分别作为Δ(5)和Δ(11)去饱和酶发挥作用。这是关于粘细菌中FA去饱和酶的异源表达和区域选择性的首次报道。

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