RETRACTED ARTICLE: Substrates specificity of tannase from Streptomyces sviceus and Lactobacillus plantarum

• Main Authors: Dan Wang, Yao Liu, Die Lv, Xueli Hu, Qiumei Zhong, Ye Zhao, Mingbo Wu
• Format: Article
• Language: English
• Published: SpringerOpen 2018-09-01
• Series: AMB Express
• Subjects: Tannase; Ester and depside bonds; Kinetics; Substrate specificity
• Online Access: http://link.springer.com/article/10.1186/s13568-018-0677-1

Abstract Tannases can catalyze the hydrolysis of galloyl ester and depside bonds of hydrolysable tannins to release gallic acid and glucose, but tannases from different species have different substrate specificities. Our prior studies found that tannase from Lactobacillus plantarum (LP-tan) performed a higher esterase activity, while the tannase from Streptomyces sviceus (SS-tan) performed a higher depsidase activity; but the molecular mechanism is not elucidated. Based on the crystal structure of LP-tan and the amino acid sequences alignment between LP-tan and SS-tan, we found that the sandwich structure formed by Ile206-substrate-Pro356 in LP-tan was replaced with Ile253-substrate-Gly384 in SS-tan, and the flap domain (amino acids: 225–247) formed in LP-tan was missed in SS-tan, while a flap-like domain (amino acids: 93–143) was found in SS-tan. In this study, we investigated the functional role of sandwich structure and the flap (flap-like) domain in the substrate specificity of tannase. Site-directed mutagenesis was used to disrupt the sandwich structure in LP-tan (P356G) and rebuilt it in SS-tan (G384P). The flap in LP-tan and the flap-like domain in SS-tan were deleted to construct the new variants. The activity assay results showed that the sandwich and the flap domain can help to catalytic the ester bonds, while the flap-like domain in SS-tan mainly worked on the depside bonds. Enzymatic characterization and kinetics data showed that the sandwich and the flap domain can help to catalytic the ester bonds, while the flap-like domain in SS-tan may worked on the depside bonds.