Characterization of a tannin acyl hydrolase from Streptomyces sviceus with substrate preference for digalloyl ester bonds

The search for new tannases with novel enzymatic properties suitable for industrial applications has been a continuous effort since the first discovery of the enzyme more than a century ago. A tannase gene (Ss-Tan) from the Gram-positive bacterium Streptomyces sviceus was identified, chemically synt...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2015-03, Vol.99 (6), p.2663-2672
Main Authors: Wu, Mingbo, Wang, Qin, McKinstry, William J., Ren, Bin
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Analysis
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Binding sites
Biomedical and Life Sciences
Biotechnologically Relevant Enzymes and Proteins
Biotechnology
Carboxylic Ester Hydrolases - chemistry
Carboxylic Ester Hydrolases - genetics
Catalytic Domain
Chemical bonds
Chemical properties
Cloning
Cloning, Molecular
E coli
Enzyme kinetics
Enzymes
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Esters - chemistry
Genes
Genomics
Hot Temperature
Hydrogen-Ion Concentration
Hydrolases
Laboratories
Life Sciences
Microbial Genetics and Genomics
Microbiology
Molecular Sequence Data
Natural products
Peptides
Physiological aspects
Polyphenols
Protection and preservation
Protein expression
Proteins
Streptomyces
Streptomyces - enzymology
Studies
Substrate Specificity
Tannins - chemistry
Tea
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Summary:The search for new tannases with novel enzymatic properties suitable for industrial applications has been a continuous effort since the first discovery of the enzyme more than a century ago. A tannase gene (Ss-Tan) from the Gram-positive bacterium Streptomyces sviceus was identified, chemically synthesized, and cloned into a C-terminal His-tagged vector for expression in Escherichia coli . The tannase possesses the active site motif of GXSXG that is conserved for serine hydrolases. The residues that constitute the catalytic triad and galloyl binding site in bacterial tannases are found conserved in Ss-Tan, which include Ser209, Asp452, His484 and Lys370, Glu384, Asp454, respectively. Ss-Tan was overexpressed in E. coli BL21-AI cells with high productivity. Enzymatic assay revealed that the enzyme displays tannase activities to hydrolyze both the ester bonds and depside bonds in hydrolyzable tannins. Kinetic analysis indicated that the enzyme preferentially acts on depside bonds with considerably higher substrate affinity and catalytic efficiency. The enzyme showed maximum activity around pH 8.0 and at 50 °C with the highest melting temperature close to 70 °C. The high depsidase activity and thermostablility of Ss-Tan may make the enzyme suitable for potential industrial applications to achieve complete digestion of hydrolyzable tannins.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-014-6085-9