Identification and biochemical characterization of a novel exo-type beta-agarase Aga3463 from an Antarctic Pseudoalteromonas sp. strain

doi:10.1016/j.ijbiomac.2019.01.204

	Identification and biochemical characterization of a novel exo-type beta-agarase Aga3463 from an Antarctic Pseudoalteromonas sp. strain
	Li, Jiang 1,2; Xie, Maisheng 1,2; Gao, Yang 3
刊名	INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
	2019-05-15
卷号	129 页码:162-170
关键词	Antarctic bacterium Exo-type beta-agarase Neoagarobiose GH86 family
ISSN号	0141-8130
DOI	10.1016/j.ijbiomac.2019.01.204
英文摘要	An agar-degrading Antarctic bacterium was isolated and identified as Pseudoalteromonas sp. NJ21. This strain showed strong agarolytic activity and could use agar as the sole carbon source for growth. A novel beta-agarase gene aga3463 was cloned and identified from the genomic library of Pseudoalteromonas sp. NJ21. It was predicted to encode a peptide of 366 amino acids, including a 21-amino-acid signal peptide. BLAST analysis revealed the deduced amino acid sequence of aga3463 shared less than 36% similarity with any characterized agarase, and phylogenetic analysis showed it belonged to the glycoside hydrolase GH86 family. The recombinant Aga3463 protein had optimal temperature and pH of 50 degrees C and 7.0, respectively, and retained more than 60% activity from 10 to 50 degrees C. The metal ions Cd2+, Ca2+, Fe3+, and Mn2+ increased Aga3463 activity, whereas Cu2+, Si2+, Fe2+, and Ni2+ decreased Aga3463 activity. Notably, Aga3463 exhibited good thermostability in the presence of Ca2+; however, Ca2+ was not necessary for its catalytic activity. The K-m and V-max values for Aga3463 were 6.17 mg/ml and 557 U/mg, respectively. Thin layer chromatography, liquid chromatography-mass spectrometry, nuclear magnetic resonance spectroscopy, and enzyme assay using p-nitrophenyl-alpha/beta-D-galactopyranoside revealed Aga3463 as an exo-type beta-agarase with the ability to degrade agarose to neoagarobiose as the major end product. (C) 2019 Elsevier B.V. All rights reserved.
资助项目	Chinese Polar Environment Comprehensive Investigation & Assessment Programs[CHINARE2017-01-05]
WOS关键词	ENZYMATIC-PROPERTIES ; BACILLUS-SUBTILIS ; CLONING ; EXPRESSION ; BACTERIUM ; GENE ; FAMILY ; PURIFICATION ; HYDROLASES ; BINDING
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry ; Polymer Science
语种	英语
出版者	ELSEVIER SCIENCE BV
WOS记录号	WOS:000466621200018
内容类型	期刊论文
源URL	[http://ir.fio.com.cn:8080/handle/2SI8HI0U/24720]
专题	自然资源部第一海洋研究所
通讯作者	Li, Jiang
作者单位	1.State Ocean Adm, Inst Oceanog 1, Key Lab Marine Bioact Subst, Qingdao 266061, Shandong, Peoples R China 2.Key Lab Marine Nat Prod Qingdao, Qingdao 266061, Shandong, Peoples R China 3.Qingdao Univ Sci & Technol, Coll Marine Sci & Biol Engn, Qingdao 266042, Shandong, Peoples R China
推荐引用方式 GB/T 7714	Li, Jiang,Xie, Maisheng,Gao, Yang. Identification and biochemical characterization of a novel exo-type beta-agarase Aga3463 from an Antarctic Pseudoalteromonas sp. strain[J]. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,2019,129:162-170.
APA	Li, Jiang,Xie, Maisheng,&Gao, Yang.(2019).Identification and biochemical characterization of a novel exo-type beta-agarase Aga3463 from an Antarctic Pseudoalteromonas sp. strain.INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,129,162-170.
MLA	Li, Jiang,et al."Identification and biochemical characterization of a novel exo-type beta-agarase Aga3463 from an Antarctic Pseudoalteromonas sp. strain".INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 129(2019):162-170.