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Lysine ε-aminotransferases: kinetic constants, substrate specificities, and the variation in active site residues

Paper ID Volume ID Publish Year Pages File Format Full-Text
16821 42615 2016 6 PDF Available
Title
Lysine ε-aminotransferases: kinetic constants, substrate specificities, and the variation in active site residues
Abstract

•Activities of three lysine ε-aminotransferase (lysAT) were compared.•LysAT from Saccharopolyspora erythraea (lysAT_Sery) showed the highest activity among three lysATs tested.•LysAT_Sery showed activity only to l-lysine.•We found two sites having residue variation among active site residues of lysAT.

l-Lysine ε-aminotransferase (lysAT) is an important enzyme in tailoring the terminal amino group of l-lysine or l-ornithine and can be directed to the synthesis of various value-added chemicals such as adipic acid. Three lysATs, lysAT from Saccharopolyspora erythraea NRRL 2338 (lysAT_Sery), lysAT from Nocardia farcinica IFM 10152, and lysAT from Rhodococcus jostii RHA1, were cloned, and their kinetic values and substrate specificities were investigated. In the reaction using 5 mM l-lysine and 10 mM α-ketoglutarate, lysAT_Sery from S. erythraea NRRL 2338 showed 72% higher specific activity than lysAT from Nocardia farcinica IFM 10152 and 42% higher specific activity than lysAT from R. jostii RHA1. More interesting result was that lysAT Sery, exhibiting the highest activity among three lysATs, did not show any activity to l-ornithine. The alignment of 146 lysAT sequences from RefSeq database was searched by the EC number of lysAT to compare the active site residues among the lysAT sequences. The sequence alignment showed that only two residues, corresponding to Ala129 and Asn328 of lysAT from Mycobacterium tuberculosis H37Rv (lysAT_Mtub), showed variations among the active site residues. All the active site residues except those two residues were completely conserved throughout 145 lysAT sequences. lysAT from S. erythraea NRRL 2338 has A129T and N328S variations (residue numbers are those of the crystal structure of lysAT_Mtub). The structural analysis by the homology model indicate that Thr126 by A129T variation in lysAT_Sery is appeared to interact more tightly with the phosphate group of PLP than alanine (the distance between Thr126 and the phosphate group of PLP was 2.92 Å). In addition, Ser328 is located at the substrate recognition site of active site and, therefore, N328S variation may be connected to the substrate specificity of lysAT.

Keywords
Lysine ε-aminotransferase; Substrate specificity; Enzyme characterization; Kinetic constants; Sequence analysis; Structural analysis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 84, March 2016, Pages 11–16
Authors
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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