fulltext.study @t Gmail

Exploitation and characterization of three versatile amidase super family members from Delftia tsuruhatensis ZJB-05174

Paper ID Volume ID Publish Year Pages File Format Full-Text
16803 42613 2016 10 PDF Available
Title
Exploitation and characterization of three versatile amidase super family members from Delftia tsuruhatensis ZJB-05174
Abstract

•Three novel amidases were identified and exploited from Delftia tsuruhatensis.•Amidases from two families showed distinct substrate spectrum and bioproperties.•Dt-Ami 2 was reported as alkaliphilic amidase showing maximum activity at pH 11.0.•A novel bioprocess for the hydrolysis of 1-cyanocyclohexylacetamide was developed.

Amidases can be assigned into two families according to their amino acid sequences. Three amidases (Dt-Amis) were mined and identified from genome of Delftia tsuruhatensis. Homology analysis demonstrated that Dt-Ami 2 and Dt-Ami 6 belonged to amidase signature (AS) family, while Dt-Ami 7 belonged to nitrilase superfamily. AS amidases were shown to hydrolyze a wide spectrum of amides. Kinetic analysis demonstrated that the extension of chain length of aliphatic amides considerably decreased the Km values, and the turnover numbers (kcat) were high with linear aliphatic amides as substrates. Dt-Ami 2 showed maximum activity near a quite alkaline pH (11.0) and exhibited opposite enantioselectivity to Dt-Ami 6. Furthermore, a novel bioprocess for hydrolysis of 1-cyanocyclohexaneacetamide was developed using Dt-Ami 6 as biocatalyst, resulting in >99% conversion within 1.5 h at a substrate loading of 100 g/L by 0.5 g/L of Escherichia coli cells. On the other hand, nitrilase superfamily amidase only hydrolyzed aliphatic amides. The Km values of Dt-Ami 7 were considerably increased with the extension of chain length of aliphatic amides. The characterized enzymes from different families showed distinct biochemical characteristics and catalytic properties, leading to a better understanding of the two super amidase family members.

Keywords
Amidase; Amidase signature family; Nitrilase superfamily; Gene mining; Kinetic resolution; Delftia tsuruhatensis
First Page Preview
Exploitation and characterization of three versatile amidase super family members from Delftia tsuruhatensis ZJB-05174
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 86, May 2016, Pages 93–102
Authors
, , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us