Discovery and characterization of a novel protease from the Antarctic soil. (December 2021)

• Record Type: Journal Article
• Authors: Chen, Wencui; Zeng, Yinxin; Zheng, Li; Liu, Weizhi; Lyu, Qianqian
• Is Part Of: Process biochemistry. Volume 111:Part 2(2021)
• Page Start: 270
• Page End: 277
• Publication Date: 2021-12
• Subjects: Protease -- Metagenomic library -- Functional screening -- Antarctic soil; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2021.10.032
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: A Fosmid metagenomic library of Antarctic soil was successfully constructed. A novel protease, Pr18H-2, was screened using a functional metagenomic strategy. Pr18H-2 was a member of the subfamily M43B non-peptidase homologs. Pr18H-2 showed excellent pH-tolerance, as well as a broad substrate profile. Abstract: The extensive use of proteases in many areas, especially in the textile, food, and detergent industries, has led increases in the discovery and characterization of novel proteolytic enzymes. Microorganism from the Antarctic may contain novel proteases because of the extreme environment; however, to our knowledge, few relevant studies have been reported to date. Here, an Antarctic soil metagenomic fosmid library was successfully constructed, and a 36 kb positive fosmid clone was screened using lactose-free skim milk agar. Sequence alignment and phylogenetic analysis indicated that the positive fosmid clone encoded a new protein belonging to the metalloprotease subfamily M43B (36.6 % identity), which was named as Pr18H-2. Enzymatic analysis revealed that Pr18H-2 possess several features: (1). it demonstrated high thermal stability with the optimal temperature was 60 °C; (2). it was active in the broad pH range of 5.0–10.0, with the optimal pH of 8.0 which allowed it to be an alkaline protease; (3). it was able to degrade a wide range of protein substrates and exhibit the substrate preferences for small substrates, such as casein hydrolysate and fibrin; (4). it showed that the enzymatic activity was increased by approximately four times in the presence of 5 mM Co 2+ .