Proceedings of the 2016 International Conference on Biomedical and Biological Engineering

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Exploration and Research on Relieving Feedback Inhibition for Aspartate Kinase form Corynebacterium Pekinense by using Site-Saturation Mutagenesis and High Throughput Screening

Authors
Li Fang, Longyang Wang, Chunlei Liu, Zhirui Zhang, Chen Chen, Weihong Min
Corresponding Author
Li Fang
Available Online July 2016.
DOI
10.2991/bbe-16.2016.6How to use a DOI?
Keywords
Aspartate Kinase, Feedback Inhibition, Site-Saturation Mutagenesis, High Throughput Screening, Characterization of Enzymology Properties.
Abstract

This research investigated the aspartate kinase (AK) of Corynebacterium pekinense (CpAK). The amino acid residue A297, which is closely related to feedback inhibition, was selected for site-directed mutagenesis, high-throughput screening, and enzyme kinetic analysis. In this research, the enzyme activity of mutant A297K AK was enhanced. The maximum reaction rate of A297K AK increased by 9.55 times compared with that of the wild-type CpAK (WT AK). In addition, Km value increased and n value decreased. The enzymologic properties are characterized as follows: optimum reaction temperature increased from 25°C to 30°C; the optimum pH remained to be 8.0; and the stability of A297K AK under the optimal conditions extended from 3.5 h to 5 h. Feedback inhibition of low concentrations of threonine and lysine and the synergistic feedback inhibition of both amino acids were successfully relieved.

Copyright
© 2016, the Authors. Published by Atlantis Press.
Open Access
This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

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Volume Title
Proceedings of the 2016 International Conference on Biomedical and Biological Engineering
Series
Advances in Biological Sciences Research
Publication Date
July 2016
ISBN
10.2991/bbe-16.2016.6
ISSN
2468-5747
DOI
10.2991/bbe-16.2016.6How to use a DOI?
Copyright
© 2016, the Authors. Published by Atlantis Press.
Open Access
This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

Cite this article

risenwbib
TY  - CONF
AU  - Li Fang
AU  - Longyang Wang
AU  - Chunlei Liu
AU  - Zhirui Zhang
AU  - Chen Chen
AU  - Weihong Min
PY  - 2016/07
DA  - 2016/07
TI  - Exploration and Research on Relieving Feedback Inhibition for Aspartate Kinase form Corynebacterium Pekinense by using Site-Saturation Mutagenesis and High Throughput Screening
BT  - Proceedings of the 2016 International Conference on Biomedical and Biological Engineering
PB  - Atlantis Press
SP  - 28
EP  - 34
SN  - 2468-5747
UR  - https://doi.org/10.2991/bbe-16.2016.6
DO  - 10.2991/bbe-16.2016.6
ID  - Fang2016/07
ER  -