Identification and characterization of a novel alkaline alpha-amylase Amy703 belonging to a new clade from Bacillus pseudofirmus

CORC > 天津工业生物技术研究所 > 微生物功能基因组学与生物技术实验室田朝光 > 期刊论文

	Identification and characterization of a novel alkaline alpha-amylase Amy703 belonging to a new clade from Bacillus pseudofirmus
	Lu, Zhenghui 1; Tian, Chaoguang 2; Li, Aiying 3; Zhang, Guimin 1; Ma, Yanhe 2
刊名	JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
	2014-05-01
卷号	41 期号:5 页码:783-793
关键词	Alkaline alpha-amylase Enzymatic assay Metalloenzyme Structural modeling Site-directed mutagenesis
英文摘要	Alkaline alpha-amylases are of great interest in desizing processes and detergent industries. Here, an alkaline alpha-amylase gene amy703 from an alkaliphilic Bacillus pseudofirmus strain was cloned and sequenced. Its encoding product, Amy703, might represent a new clade of alpha-amylase family, because it shared only 35 % highest identity with all amylases characterized up to date and was not clustered into any subfamilies with amylase activity in glycoside hydrolase family 13. Heterologous expression and characterization of Amy703 showed that it is a metalloenzyme with maximal activity at 40 A degrees C and pH 9.0. Its activity was significantly enhanced by 2- and 2.48-fold at the presence of 10 mM Ca2+ and Mg2+, respectively, while Hg2+ was a strong inhibitor of Amy703. Amy703 has a higher affinity (K (m) = 3.92 mg/ml) for soluble starch compared to many other alkaline amylases. The computer modeling of its structure indicated that Amy703 contains typical amylase domains and a loop region appearing to bind the substrates. Site-directed mutagenesis suggested that a conserved residue Glu550 was essential for the activity of Amy703, and proposed it working together with other two residues to constitute a catalytic triad (Asp521, Glu550, and Asp615).
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology
关键词[WOS]	BIOCHEMICAL-CHARACTERIZATION ; HETEROLOGOUS EXPRESSION ; CLONING ; SUBTILIS ; PURIFICATION ; HYDROLYSIS ; RESOLUTION ; RESISTANT ; RESIDUES ; MALTOSE
收录类别	SCI
语种	英语
WOS记录号	WOS:000334423400004
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1394]
专题	天津工业生物技术研究所_微生物功能基因组学与生物技术实验室田朝光_期刊论文
作者单位	1.Hubei Univ, Coll L ife Sci, Hubei Collaborat Innovat Ctr Green Transformat Bi, Wuhan 430062, Peoples R China 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China 3.Cent China Normal Univ, Coll Life Sci, Wuhan 430079, Peoples R China
推荐引用方式 GB/T 7714	Lu, Zhenghui,Tian, Chaoguang,Li, Aiying,et al. Identification and characterization of a novel alkaline alpha-amylase Amy703 belonging to a new clade from Bacillus pseudofirmus[J]. JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY,2014,41(5):783-793.
APA	Lu, Zhenghui,Tian, Chaoguang,Li, Aiying,Zhang, Guimin,&Ma, Yanhe.(2014).Identification and characterization of a novel alkaline alpha-amylase Amy703 belonging to a new clade from Bacillus pseudofirmus.JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY,41(5),783-793.
MLA	Lu, Zhenghui,et al."Identification and characterization of a novel alkaline alpha-amylase Amy703 belonging to a new clade from Bacillus pseudofirmus".JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY 41.5(2014):783-793.