Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>

Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>

This report deals with the purification, characterization, and a preliminary crystallographic study of a novel cold-active esterase (<i>Ha</i>Est1) from Halocynthiibacter arcticus. Primary sequence analysis reveals that <i>Ha</i>Est1 has a catalytic serine in G-x-S-x-G motif....

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Main Authors: Sangeun Jeon, Jisub Hwang, Wanki Yoo, Joo Won Chang, Hackwon Do, Han-Woo Kim, Kyeong Kyu Kim, Jun Hyuck Lee, T. Doohun Kim
Format: Article
Language:English
Published: MDPI AG 2021-02-01
Series:Crystals
Subjects:
esterase
enzyme assay
crystallization
diffraction
Online Access:https://www.mdpi.com/2073-4352/11/2/170
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spelling doaj-f2a77e97f5d24541a7b654f4116876502021-02-09T00:04:56ZengMDPI AGCrystals2073-43522021-02-011117017010.3390/cryst11020170Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>Sangeun Jeon0Jisub Hwang1Wanki Yoo2Joo Won Chang3Hackwon Do4Han-Woo Kim5Kyeong Kyu Kim6Jun Hyuck Lee7T. Doohun Kim8Department of Chemistry, Graduate School of General Studies, Sookmyung Women’s University, Seoul 04310, KoreaResearch Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, KoreaDepartment of Chemistry, Graduate School of General Studies, Sookmyung Women’s University, Seoul 04310, KoreaDepartment of Chemistry, Graduate School of General Studies, Sookmyung Women’s University, Seoul 04310, KoreaResearch Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, KoreaResearch Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, KoreaDepartment of Molecular Cell Biology, School of Medicine, Sungkyunkwan University, Suwon 16419, KoreaResearch Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, KoreaDepartment of Chemistry, Graduate School of General Studies, Sookmyung Women’s University, Seoul 04310, KoreaThis report deals with the purification, characterization, and a preliminary crystallographic study of a novel cold-active esterase (<i>Ha</i>Est1) from Halocynthiibacter arcticus. Primary sequence analysis reveals that <i>Ha</i>Est1 has a catalytic serine in G-x-S-x-G motif. The recombinant <i>Ha</i>Est1 was cloned, expressed, and purified. SDS-PAGE and zymographic analysis were carried out to characterize the properties of <i>Ha</i>Est1. A single crystal of <i>Ha</i>Est1 was obtained in a solution containing 10% (w/v) PEG 8000/8% ethylene glycol, 0.1 M Hepes-NaOH, pH 7.5. Diffraction data were collected to 2.10 Å resolution with P2<sub>1</sub> space group. The final R<sub>merge</sub> and R<sub>p.i.m</sub> values were 7.6% and 3.5% for 50-2.10 Å resolution. The unit cell parameters were <i>a</i> = 35.69 Å, <i>b</i> = 91.21 Å, <i>c</i> = 79.15 Å, and = 96.9°.https://www.mdpi.com/2073-4352/11/2/170esteraseenzyme assaycrystallizationdiffraction
collection DOAJ
language English
format Article
sources DOAJ
author Sangeun Jeon
Jisub Hwang
Wanki Yoo
Joo Won Chang
Hackwon Do
Han-Woo Kim
Kyeong Kyu Kim
Jun Hyuck Lee
T. Doohun Kim
spellingShingle Sangeun Jeon
Jisub Hwang
Wanki Yoo
Joo Won Chang
Hackwon Do
Han-Woo Kim
Kyeong Kyu Kim
Jun Hyuck Lee
T. Doohun Kim
Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>
Crystals
esterase
enzyme assay
crystallization
diffraction
author_facet Sangeun Jeon
Jisub Hwang
Wanki Yoo
Joo Won Chang
Hackwon Do
Han-Woo Kim
Kyeong Kyu Kim
Jun Hyuck Lee
T. Doohun Kim
author_sort Sangeun Jeon
title Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>
title_short Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>
title_full Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>
title_fullStr Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>
title_full_unstemmed Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (<i>Ha</i>Est1) from <i>Halocynthiibacter arcticus</i>
title_sort purification and crystallographic analysis of a novel cold-active esterase (<i>ha</i>est1) from <i>halocynthiibacter arcticus</i>
publisher MDPI AG
series Crystals
issn 2073-4352
publishDate 2021-02-01
description This report deals with the purification, characterization, and a preliminary crystallographic study of a novel cold-active esterase (<i>Ha</i>Est1) from Halocynthiibacter arcticus. Primary sequence analysis reveals that <i>Ha</i>Est1 has a catalytic serine in G-x-S-x-G motif. The recombinant <i>Ha</i>Est1 was cloned, expressed, and purified. SDS-PAGE and zymographic analysis were carried out to characterize the properties of <i>Ha</i>Est1. A single crystal of <i>Ha</i>Est1 was obtained in a solution containing 10% (w/v) PEG 8000/8% ethylene glycol, 0.1 M Hepes-NaOH, pH 7.5. Diffraction data were collected to 2.10 Å resolution with P2<sub>1</sub> space group. The final R<sub>merge</sub> and R<sub>p.i.m</sub> values were 7.6% and 3.5% for 50-2.10 Å resolution. The unit cell parameters were <i>a</i> = 35.69 Å, <i>b</i> = 91.21 Å, <i>c</i> = 79.15 Å, and = 96.9°.
topic esterase
enzyme assay
crystallization
diffraction
url https://www.mdpi.com/2073-4352/11/2/170
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