Abstract
Only few enzymes from haloalkaliphiles are biochemically characterized for their kinetic behaviour and stability. In view of this realization, an alkaline protease from Bacillus sp. AH-6, displaying salt-dependent resistance against chemical denaturation by Urea and Guanidium hydrochloride was investigated for denaturation and in vitro protein folding. The crude enzyme was highly resistant against urea (8 M) denaturation up to 72 h; however, on purification, it turned sensitive and got denatured within 2 h. Interestingly, the purified enzyme regained the resistance in the presence of NaCl. Effective refolding of the purified enzyme was achieved with glycerol; however, other approaches such as lower protein concentrations, rapid dilution and slow removal of the denaturant did not further add to refolding. The results are important from the viewpoint that only few enzymes from haloalkaliphilic bacteria are characterized. Since the resistance against chemical denaturation is a rare phenomenon, the findings would enrich the knowledge on protein stability and denaturation. Besides, such biocatalysts would definitely have novel applications under harsh chemical environments.
References
Mar 19, 1991·Biochemistry·M E GoldbergR Jaenicke
Sep 25, 1995·FEBS Letters·V DötschK Wüthrich
Feb 1, 1995·Protein Engineering·Y MaedaT Imoto
Jan 1, 1996·Protein Engineering·Y MaedaT Imoto
Dec 16, 1997·Biophysical Chemistry·R FavillaA Mazzini
May 16, 2002·Extremophiles : Life Under Extreme Conditions·Yaroslava Y PolosinaOleg V Fedorov
Mar 22, 2003·Protein Science : a Publication of the Protein Society·Yuji O KamatariTakashi Konno
Jun 15, 2004·Biochimie·Dominique Madern, Giuseppe Zaccai
Jun 29, 2004·Extremophiles : Life Under Extreme Conditions·Dominique MadernGiuseppe Zaccai
Sep 4, 2004·Biophysical Journal·Brian M Baynes, Bernhardt L Trout
Feb 8, 2005·Journal of Colloid and Interface Science·E AbuinC Solar
Feb 8, 2005·The Journal of Biological Chemistry·Rajesh MishraRajiv Bhat
Jun 25, 2005·Journal of Chromatography. a·Anshu GuptaM N Gupta
Jul 20, 2006·Critical Reviews in Biochemistry and Molecular Biology·Zong Lin, Hays S Rye
May 15, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Subhankar PaulTapan K Chaudhuri
Citations
Jul 24, 2013·Extremophiles : Life Under Extreme Conditions·Vikash Kumar, T Satyanarayana
Feb 14, 2012·Applied Biochemistry and Biotechnology·Sandeep Pandey, S P Singh
Dec 11, 2013·BioMed Research International·Darine El HidriAmeur Cherif
Nov 6, 2014·Canadian Journal of Microbiology·Baisuo ZhaoShulin Chen
Nov 28, 2012·International Journal of Biological Macromolecules·Megha K Purohit, Satya P Singh
Dec 20, 2011·Bioresource Technology·Nadia Zaraî JaouadiSamir Bejar
Jul 11, 2014·The Protein Journal·Rajeshwari Sinha, S K Khare
Aug 29, 2009·Letters in Applied Microbiology·M K Purohit, S P Singh
Jun 10, 2011·Journal of Basic Microbiology·Anil Kumar Singh, Hari S Chhatpar
Jul 15, 2015·Extremophiles : Life Under Extreme Conditions·Abdelnasser S S IbrahimGarabed Antranikian
Feb 9, 2013·International Journal of Biological Macromolecules·S D Gohel, S P Singh
May 2, 2014·Frontiers in Microbiology·Rajeshwari Sinha, Sunil K Khare
Jan 31, 2017·The Journal of Physical Chemistry. B·Nicola SteinkeSylvia E McLain
Oct 28, 2019·3 Biotech·Abdelnasser S S IbrahimAtif A Elagib
Jan 18, 2020·Biotechnology Progress·Amal HammamiMoncef Nasri
Apr 24, 2020·World Journal of Microbiology & Biotechnology·Ganapathi UmaThavasimuthu Citarasu
Jun 26, 2020·Frontiers in Microbiology·Hitarth B Bhatt, Satya P Singh
Jul 28, 2020·Genomics·Nirali M Raiyani, Satya P Singh
Apr 23, 2011·International Journal of Biological Macromolecules·Megha K Purohit, Satya P Singh