Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (HaEst1) from Halocynthiibacter arcticus
  • Jeon, Sangeun
  • Hwang, Jisub
  • Yoo, Wanki
  • Chang, Joo Won
  • Do, Hackwon
  • 외 4명
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초록

This report deals with the purification, characterization, and a preliminary crystallographic study of a novel cold-active esterase (HaEst1) from Halocynthiibacter arcticus. Primary sequence analysis reveals that HaEst1 has a catalytic serine in G-x-S-x-G motif. The recombinant HaEst1 was cloned, expressed, and purified. SDS-PAGE and zymographic analysis were carried out to characterize the properties of HaEst1. A single crystal of HaEst1 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 angstrom resolution with P2(1) space group. The final R-merge and R-p.i.m values were 7.6% and 3.5% for 50-2.10 angstrom resolution. The unit cell parameters were a = 35.69 angstrom, b = 91.21 angstrom, c = 79.15 angstrom, and beta = 96.9 degrees.

키워드

esteraseenzyme assaycrystallizationdiffraction
제목
Purification and Crystallographic Analysis of a Novel Cold-Active Esterase (HaEst1) from Halocynthiibacter arcticus
저자
Jeon, SangeunHwang, JisubYoo, WankiChang, Joo WonDo, HackwonKim, Han-WooKim, Kyeong KyuLee, Jun HyuckKim, T. Doohun
DOI
10.3390/cryst11020170
발행일
2021-02
유형
Article
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