干旱区生态环境知识资源中心(Arid): Cloning, Expression, and Structural Elucidation of a Biotechnologically Potential Alkaline Serine Protease From a Newly Isolated HaloalkaliphilicBacillus lehensisJO-26

Arid

DOI	10.3389/fmicb.2020.00941
	Cloning, Expression, and Structural Elucidation of a Biotechnologically Potential Alkaline Serine Protease From a Newly Isolated HaloalkaliphilicBacillus lehensisJO-26
	Bhatt, Hitarth B.; Singh, Satya P.
通讯作者	Singh, Satya P.
来源期刊	FRONTIERS IN MICROBIOLOGY
ISSN	1664-302X
出版年	2020
卷号	11
英文摘要	An alkaline protease gene ofBacillus lehensisJO-26 from saline desert, Little Rann of Kutch, was cloned and expressed inEscherichia coliBL21 (DE3). A 1,014-bp ORF encoded 337 amino acids. The recombinant protease (APrBL) with Asp 97, His 127, and Ser 280 forming catalytic triad belongs to the subtilase S8 protease family. The gene was optimally expressed in soluble fraction with 0.2 mM isopropyl beta-D-thiogalactopyranoside (IPTG), 2% (w/v) NaCl at 28 degrees C. APrBL, a monomer with a molecular mass of 34.6 kDa was active over pH 8-11 and 30 degrees C-70 degrees C, optimally at pH 10 and 50 degrees C. The enzyme was highly thermostable and retained 73% of the residual activity at 80 degrees C up to 3 h. It was significantly stimulated by sodium dodecyl sulfate (SDS), Ca2+, chloroform, toluene, n-butanol, and benzene while completely inhibited by phenylmethylsulfonyl fluoride (PMSF) and Hg2+. The serine nature of the protease was confirmed by its strong inhibition by PMSF. The APrBL gene was phylogenetically close to alkaline elastase YaB (P20724) and was distinct from the well-known commercial proteases subtilisin Carlsberg (CAB56500) and subtilisin BPN ' (P00782). The structural elucidation revealed 31.75% alpha-helices, 22.55% beta-strands, and 45.70% coils. Although high glycine and fewer proline residues are a characteristic feature of the cold-adapted enzymes, the similar observation in thermally active APrBL suggests that this feature cannot be solely responsible for thermo/cold adaptation. The APrBL protease was highly effective as a detergent additive and in whey protein hydrolysis.
英文关键词	recombinant alkaline protease gene expression Little Rann of Kutch structure-function relationship detergent additive whey protein hydrolysis
类型	Article
语种	英语
国家	India
开放获取类型	Green Published, gold
收录类别	SCI-E
WOS记录号	WOS:000543320700001
WOS关键词	BACILLUS SP ; HALOALKALIPHILIC BACTERIUM ; MOLECULAR CHARACTERIZATION ; OVER-EXPRESSION ; PURIFICATION ; SUBTILISIN ; STABILITY ; TEMPERATURE ; STRATEGIES ; ENZYMES
WOS类目	Microbiology
WOS研究方向	Microbiology
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/319545
作者单位	Saurashtra Univ, UGC CAS Dept Biosci, Rajkot, Gujarat, India
推荐引用方式 GB/T 7714	Bhatt, Hitarth B.,Singh, Satya P.. Cloning, Expression, and Structural Elucidation of a Biotechnologically Potential Alkaline Serine Protease From a Newly Isolated HaloalkaliphilicBacillus lehensisJO-26[J],2020,11.
APA	Bhatt, Hitarth B.,&Singh, Satya P..(2020).Cloning, Expression, and Structural Elucidation of a Biotechnologically Potential Alkaline Serine Protease From a Newly Isolated HaloalkaliphilicBacillus lehensisJO-26.FRONTIERS IN MICROBIOLOGY,11.
MLA	Bhatt, Hitarth B.,et al."Cloning, Expression, and Structural Elucidation of a Biotechnologically Potential Alkaline Serine Protease From a Newly Isolated HaloalkaliphilicBacillus lehensisJO-26".FRONTIERS IN MICROBIOLOGY 11(2020).