Exploration and Research on Relieving Feedback Inhibition for Aspartate Kinase form Corynebacterium Pekinense by using Site-Saturation Mutagenesis and High Throughput Screening
Li Fang, Longyang Wang, Chunlei Liu, Zhirui Zhang, Chen Chen, Weihong Min
Available Online July 2016.
- https://doi.org/10.2991/bbe-16.2016.6How to use a DOI?
- Aspartate Kinase, Feedback Inhibition, Site-Saturation Mutagenesis, High Throughput Screening, Characterization of Enzymology Properties.
- 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.
- Open Access
- This is an open access article distributed under the CC BY-NC license.
Cite this article
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 - https://doi.org/10.2991/bbe-16.2016.6 ID - Fang2016/07 ER -