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来自芽孢杆菌属K91的一种热稳定碱性GDSL基序酯酶：结晶与X射线晶体学分析

A thermostable and alkaline GDSL-motif esterase from Bacillus sp. K91: crystallization and X-ray crystallographic analysis.

作者信息

Ding Junmei, Zhu Hujie, Ye Yujia, Li Jie, Han Nanyu, Wu Qian, Huang Zunxi, Meng Zhaohui

机构信息

Engineering Research Centre of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, 768 Juxian Road, Kunming, Yunnan 650500, People's Republic of China.

Laboratory of Molecular Cardiology, Department of Cardiology, The First Affiliated Hospital of Kunming Medical University, 1168 Chunrong West Road, Kunming, Yunnan 650500, People's Republic of China.

出版信息

Acta Crystallogr F Struct Biol Commun. 2018 Feb 1;74(Pt 2):117-121. doi: 10.1107/S2053230X18000353. Epub 2018 Jan 26.

DOI:10.1107/S2053230X18000353
PMID:29400322
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5947683/
Abstract

The esterase Est8 from the thermophilic bacterium Bacillus sp. K91 belongs to the GDSL family and is active on a variety of acetylated compounds, including 7-aminocephalosporanic acid. In contrast to other esterases of the GDSL family, the catalytic residues Asp182 and His185 were more pivotal for the catalytic activity of Est8 than the Ser11 residue. To better understand the biochemical and enzymatic properties of Est8, recombinant Est8 protein was purified and crystallized. Crystals of Est8 were obtained by the hanging-drop vapour-diffusion method using 2.0 M ammonium sulfate, 5%(v/v) 2-propanol as the crystallization solution. X-ray diffraction data were collected to a resolution of 2.30 Å with an R of 16.4% from a crystal belonging to space group P422 or P422, with unit-cell parameters a = b = 68.50, c = 79.57 Å.

摘要

嗜热芽孢杆菌Bacillus sp. K91中的酯酶Est8属于GDSL家族，对多种乙酰化化合物具有活性，包括7-氨基头孢烷酸。与GDSL家族的其他酯酶不同，催化残基Asp182和His185对Est8的催化活性比Ser11残基更为关键。为了更好地了解Est8的生化和酶学性质，对重组Est8蛋白进行了纯化和结晶。使用2.0 M硫酸铵、5%（v/v）异丙醇作为结晶溶液，通过悬滴气相扩散法获得了Est8晶体。从属于空间群P422或P422的晶体收集了分辨率为2.30 Å的X射线衍射数据，R值为16.4%，晶胞参数a = b = 68.50，c = 79.57 Å。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/ea8e30ffcc74/f-74-00117-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/54fccb59703b/f-74-00117-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/8fe2a28fe54e/f-74-00117-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/2a14a30d2b1d/f-74-00117-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/ea8e30ffcc74/f-74-00117-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/54fccb59703b/f-74-00117-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/8fe2a28fe54e/f-74-00117-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/2a14a30d2b1d/f-74-00117-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ecf/5947683/ea8e30ffcc74/f-74-00117-fig4.jpg

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