Cloning and biochemical characterization of a novel kappa-carrageenase from newly isolated marine bacterium Pedobacter hainanensis NJ-02

doi:10.1016/j.ijbiomac.2017.11.040

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	Cloning and biochemical characterization of a novel kappa-carrageenase from newly isolated marine bacterium Pedobacter hainanensis NJ-02
	Zhu, Benwei 1; Ni, Fang 1; Ning, Limin 2; Yao, Zhong 1; Du, Yuguang 3
	2018-03-01
发表期刊	INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
ISSN	0141-8130
卷号	108 页码:1331-1338
摘要	Enzymatic preparation of carrageenan oligosaccharides has drawn increasing attention due to its advantages of mild reaction conditions and excellent product-specificity. A novel gene (CgkA) encoding a new kappa-carrageenase was cloned, heterogeneously expressed and characterized from a newly isolated marine bacterium Pedobacter hainanensis NJ-02. It consisted of 1539 bp and encoded 512 amino acid residues with a molecular weight of 57.12 kDa. Multiple alignment analysis indicated that CgkA belongs to glycoside hydrolase (GH) family 16 and was most homologous to kappa-carrageenase of Zobellia sp. M-2 with identity of 50%. The recombinant enzyme showed maximal activity of 3659.72 U/mg at 40 degrees C and pH 7.0. Additionally, it could retain more than 80% of its maximal activity after being incubated at pH of 5.0-9.0 below 40 degrees C. ICE and Na+ with a wide range of concentration can activate the enzyme, while other divalent ions such as Cu2+, Zn2+ showed inhibitory effect on the enzyme. The ESI-MS analysis of hydrolysates indicated that the enzyme can endolytically depolymerize the carrageenan into tetrasaccharides and hexasaccharides. The results suggest that it is an endo-type carrageenase and could be a valuable enzyme tool to produce carrageenan oligosaccharides with higher Dps. (C) 2017 Elsevier B.V. All rights reserved.
关键词	Carrageenase Characterization Heterologous Expression
文章类型	Article
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
DOI	10.1016/j.ijbiomac.2017.11.040
收录类别	SCI
语种	英语
关键词[WOS]	GLYCOSIDE HYDROLASES ; PURIFICATION ; OLIGOSACCHARIDES ; DEGRADATION ; PRODUCTS
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry ; Polymer Science
WOS类目	Biochemistry & Molecular Biology ; Chemistry, Applied ; Polymer Science
项目资助者	National Natural Science Foundation of China(31601410) ; National Basic Research Program of China (973 Program)(2013CB733503) ; Key Research & Development Program of Jiangsu Province(BE2015305)
WOS记录号	WOS:000425071500151
引用统计
文献类型	期刊论文
条目标识符	http://ir.ipe.ac.cn/handle/122111/24026
专题	生化工程国家重点实验室
作者单位	1.Nanjing Tech Univ, Coll Food Sci & Light Ind, Nanjing 211816, Jiangsu, Peoples R China 2.Nanjing Univ Chinese Med, Coll Med & Life Sci, Nanjing 210023, Jiangsu, Peoples R China 3.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100190, Peoples R China
推荐引用方式 GB/T 7714	Zhu, Benwei,Ni, Fang,Ning, Limin,et al. Cloning and biochemical characterization of a novel kappa-carrageenase from newly isolated marine bacterium Pedobacter hainanensis NJ-02[J]. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,2018,108:1331-1338.
APA	Zhu, Benwei,Ni, Fang,Ning, Limin,Yao, Zhong,&Du, Yuguang.(2018).Cloning and biochemical characterization of a novel kappa-carrageenase from newly isolated marine bacterium Pedobacter hainanensis NJ-02.INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,108,1331-1338.
MLA	Zhu, Benwei,et al."Cloning and biochemical characterization of a novel kappa-carrageenase from newly isolated marine bacterium Pedobacter hainanensis NJ-02".INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 108(2018):1331-1338.