Cold-adaptive alkaline protease from the psychrophilic Planomicrobium sp. 547: enzyme characterization and gene cloning

Xiangsheng, Yang and Xinglin, Chen and Xianzhong, Xu and Runying, Zheng (2011) Cold-adaptive alkaline protease from the psychrophilic Planomicrobium sp. 547: enzyme characterization and gene cloning. Advances in Polar Science, 22 (1). pp. 49-54.

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Abstract

A psychrophilic bacterium strain 547 producing cold-adaptive alkaline protease was isolated from the deep sea sediment of Prydz Bay, Antarctica. The organism was identified as a Planomicrobium species by 16S rRNA analysis. The optimal and highest growth temperatures for strain 547 were 15℃ and 30℃, respectively. The extracellular protease was purified by ammonium sulfate precipitation and DEAE cellulose-52 chromatography. The optimal temperature and pH for the activity of the purified enzyme were 35℃ and pH 9.0, respectively. The enzyme retained approximately 40% of its activity after 2 h of incubation at 50℃. The enzymatic activity was inhibited by 1 mmol/L phenylmethyl sulfonylfluoride (PMSF) and hydrochloride 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), indicating that it was a serine protease. The presence of Ca2+ and Mn2+ increased the activity of the enzyme. The protease gene with a size of 1 269 bp was cloned from Planomicrobium sp. 547 using primers designed based on the conserved sequences of proteases in GenBank. The Planomicrobium sp. 547 protease contained a domain belonging to the peptidase S8 family, which has a length of 309 amino acid (AA) residues. The alignment and phylogenetic analysis of the AA sequence indicated that the protease belonged to the subtilisin family.

Item Type:	Article
Related URLs:
Keywords:	cold-adaptive protease, Planomicrobium, Antarctic, subtilisin
Subjects:	Natural Environment > Fauna
Organizations:	Unspecified
Date Deposited:	13 Sep 2023 10:49
URI:	http://library.arcticportal.org/id/eprint/2412

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