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Cloning, expression, and characterization of the beta-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1
Alternative TitleAppl. Microbiol. Biotechnol.
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
2014-03-01
Source PublicationAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN0175-7598
Volume98Issue:6Pages:2519-2531
AbstractPreviously, 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.; 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.
KeywordBacteroides Uniformis Beta-glucosidase Secoisolariciresinol Diglucoside Secoisolariciresinol Recombinant Bacteria
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1007/s00253-013-5111-7
URL查看原文
Indexed BySCI
Language英语
WOS KeywordHUMAN INTESTINAL BACTERIA ; ESCHERICHIA-COLI ; PROTEIN-PRODUCTION ; SEQUENCE-ANALYSIS ; PURIFICATION ; ENTERODIOL ; ENTEROLACTONE ; STRATEGIES ; BIOTRANSFORMATION ; IMPROVEMENT
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000332108100014
Citation statistics
Cited Times:10[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/8049
Collection研究所(批量导入)
Affiliation1.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
Recommended Citation
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.
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