Cloning, expression, and characterization of the beta-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1

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	Cloning, expression, and characterization of the beta-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1
	Tao, Y. -L.1; Yang, D. -H.1; Zhang, Y. -T.1; Zhang, Y.1; Wang, Z. -Q.2; Wang, Y. -S.2; Cai, S. -Q.1; Liu, S. -L.3,4
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2014-03-01
卷号	98 期号:6 页码:2519-2531
关键词	Bacteroides uniformis beta-glucosidase Secoisolariciresinol diglucoside Secoisolariciresinol Recombinant bacteria
ISSN号	0175-7598
其他题名	Appl. Microbiol. Biotechnol.
中文摘要	Previously, from the human intestinal flora we isolated the bacterial strain Bacteroides uniformis ZL1, which could convert secoisolariciresinol diglucoside (SDG) to its aglycone secoisolariciresinol (SECO) in vivo. In this study, 24 putative beta-glucosidase genes were screened from the genome of B. uniformis ATCC 8492, which were used as templates to design PCR primers for the target genes in B. uniformis ZL1. Fifteen genes (bgl1-bgl15) were amplified from strain ZL1, and among them we identified bgl8 as the gene encoding the SDG-hydrolyzing beta-glucosidase. We sequenced the bgl8 gene, cloned it into the expression vector and then transformed Escherichia coli to construct the recombinant bacteria that could synthesize the target beta-glucosidase (BuBGL8). We purified and characterized BuBGL8, which showed maximal activity and stability under the culture conditions of pH 6.0 and 30 A degrees C. SDG (2.0 mg/ml) was converted to SECO by both the purified BuBGL8 (0.035 mg/ml) and crude enzyme extract (0.23 mg crude protein/ml) with the efficiency of more than 90 % after 90 min at the reaction conditions. This is, to our knowledge, the first report of using recombinant bacteria to synthesize the SDG-hydrolyzing beta-glucosidase, which could be used to produce SECO from SDG conveniently and highly efficiently.
英文摘要	Previously, from the human intestinal flora we isolated the bacterial strain Bacteroides uniformis ZL1, which could convert secoisolariciresinol diglucoside (SDG) to its aglycone secoisolariciresinol (SECO) in vivo. In this study, 24 putative beta-glucosidase genes were screened from the genome of B. uniformis ATCC 8492, which were used as templates to design PCR primers for the target genes in B. uniformis ZL1. Fifteen genes (bgl1-bgl15) were amplified from strain ZL1, and among them we identified bgl8 as the gene encoding the SDG-hydrolyzing beta-glucosidase. We sequenced the bgl8 gene, cloned it into the expression vector and then transformed Escherichia coli to construct the recombinant bacteria that could synthesize the target beta-glucosidase (BuBGL8). We purified and characterized BuBGL8, which showed maximal activity and stability under the culture conditions of pH 6.0 and 30 A degrees C. SDG (2.0 mg/ml) was converted to SECO by both the purified BuBGL8 (0.035 mg/ml) and crude enzyme extract (0.23 mg crude protein/ml) with the efficiency of more than 90 % after 90 min at the reaction conditions. This is, to our knowledge, the first report of using recombinant bacteria to synthesize the SDG-hydrolyzing beta-glucosidase, which could be used to produce SECO from SDG conveniently and highly efficiently.
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]	HUMAN INTESTINAL BACTERIA ; ESCHERICHIA-COLI ; PROTEIN-PRODUCTION ; SEQUENCE-ANALYSIS ; PURIFICATION ; ENTERODIOL ; ENTEROLACTONE ; STRATEGIES ; BIOTRANSFORMATION ; IMPROVEMENT
收录类别	SCI
原文出处	://WOS:000332108100014
语种	英语
WOS记录号	WOS:000332108100014
公开日期	2014-05-06
内容类型	期刊论文
版本	出版稿
源URL	[http://ir.ipe.ac.cn/handle/122111/8049]
专题	过程工程研究所_研究所（批量导入）
作者单位	1.Peking Univ, Sch Pharmaceut Sci, State Key Lab Nat & Biomimet Drugs, Beijing 100191, Peoples R China 2.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 3.Harbin Med Univ, Genom Res Ctr, Harbin 150081, Peoples R China 4.Univ Calgary, Dept Microbiol & Infect Dis, Calgary, AB T2N 4N1, Canada
推荐引用方式 GB/T 7714	Tao, Y. -L.,Yang, D. -H.,Zhang, Y. -T.,et al. Cloning, expression, and characterization of the beta-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2014,98(6):2519-2531.
APA	Tao, Y. -L..,Yang, D. -H..,Zhang, Y. -T..,Zhang, Y..,Wang, Z. -Q..,...&Liu, S. -L..(2014).Cloning, expression, and characterization of the beta-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,98(6),2519-2531.
MLA	Tao, Y. -L.,et al."Cloning, expression, and characterization of the beta-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 98.6(2014):2519-2531.