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参与菜豆毒素生物合成的脒基转移酶AmtA的特性分析

Characterization of AmtA, an amidinotransferase involved in the biosynthesis of phaseolotoxins.

作者信息

Li Mi, Chen Li, Deng Zixin, Zhao Changming

机构信息

Key Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education School of Pharmaceutical Sciences Wuhan University China.

出版信息

FEBS Open Bio. 2016 May 16;6(6):603-9. doi: 10.1002/2211-5463.12071. eCollection 2016 Jun.

DOI:10.1002/2211-5463.12071
PMID:27419063
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4887976/
Abstract

Phaseolotoxins (PHTs), which are produced by Pseudomonas, belong to a family of phosphoramidate natural products. Two nonproteinogenic amino acid precursors, N(δ)(N'-sulfo-diaminophosphinyl)-ornithine (PSOrn) and homoarginine (hArg), are involved in biosynthesis of PHTs. Amidinotransferase AmtA catalyses the formation of hArg, with arginine and lysine as substrates. AmtA was overexpressed and purified in an Escherichia coli system. An in vitro enzyme assay showed that it has stricter substrate specificity than certain other amidinotransferases. Site-directed mutagenesis experiments showed that the mutation AmtA Met243His244 is an alternative while Met246 is essential for the transamidination activity.

摘要

菜豆毒素（PHTs）由假单胞菌产生，属于氨基磷酸酯天然产物家族。两种非蛋白质ogenic氨基酸前体，N（δ）（N'-磺基-二氨基磷酰基）-鸟氨酸（PSOrn）和高精氨酸（hArg），参与PHTs的生物合成。脒基转移酶AmtA催化以精氨酸和赖氨酸为底物形成hArg。AmtA在大肠杆菌系统中过表达并纯化。体外酶分析表明，它比某些其他脒基转移酶具有更严格的底物特异性。定点诱变实验表明，突变体AmtA Met243His244是一种替代物，而Met246对于转脒基活性至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/32d1a3fa58a6/FEB4-6-603-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/c9df4ffdf6b7/FEB4-6-603-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/7e43745e4228/FEB4-6-603-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/42520bcd61e2/FEB4-6-603-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/5ff56e811066/FEB4-6-603-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/fa6d2ac86fbb/FEB4-6-603-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/1aa11abb674c/FEB4-6-603-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/32d1a3fa58a6/FEB4-6-603-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/c9df4ffdf6b7/FEB4-6-603-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/7e43745e4228/FEB4-6-603-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/42520bcd61e2/FEB4-6-603-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/5ff56e811066/FEB4-6-603-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/fa6d2ac86fbb/FEB4-6-603-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/1aa11abb674c/FEB4-6-603-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8885/4887976/32d1a3fa58a6/FEB4-6-603-g007.jpg

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