Evolution of a chimeric aspartate kinase for L-lysine production using a synthetic RNA device

Verfasser / Beitragende:
[Junming Wang, Dongfang Gao, Xiaoli Yu, Wen Li, Qingsheng Qi]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/20(2015-10-01), 8527-8536
Format:
Artikel (online)
ID: 605499624
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024 7 0 |a 10.1007/s00253-015-6615-0  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6615-0 
245 0 0 |a Evolution of a chimeric aspartate kinase for L-lysine production using a synthetic RNA device  |h [Elektronische Daten]  |c [Junming Wang, Dongfang Gao, Xiaoli Yu, Wen Li, Qingsheng Qi] 
520 3 |a Aspartate kinase (AK) is a key enzyme involved in catalyzing the first step of the aspartate-derived amino acid biosynthesis, including L-lysine and L-threonine, which is regulated by the end-metabolites through feedback inhibition. In order to accumulate the end-metabolites in the host, the feedback inhibition of AK has to be released. In this study, a chimeric aspartate kinase, which is composed of the N-terminal catalytic region from Bacillus subtilis AKII and the C-terminal region from Thermus thermophilus, was evolved through random mutagenesis and then screened using a high-throughput synthetic RNA device which comprises of an L-lysine-sensing riboswitch and a selection module. Of three evolved aspartate kinases, the best mutant BT3 showed 160% increased in vitro activity compared to the wild-type enzyme. Recombinant Escherichia coli harboring BT3 produced 674mg/L L-lysine in batch cultivation, similar to that produced by the strain harboring the typical commercial widely used feedback resistant aspartate kinase AKC fbr from E. coli. The results suggested that this strategy can be extended for screening of other key enzymes involved in lysine biosynthesis pathways. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Riboswitch  |2 nationallicence 
690 7 |a L-lysine  |2 nationallicence 
690 7 |a Evolution  |2 nationallicence 
690 7 |a Aspartate kinase  |2 nationallicence 
690 7 |a Escherichia coli  |2 nationallicence 
700 1 |a Wang  |D Junming  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
700 1 |a Gao  |D Dongfang  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
700 1 |a Yu  |D Xiaoli  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
700 1 |a Li  |D Wen  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
700 1 |a Qi  |D Qingsheng  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/20(2015-10-01), 8527-8536  |x 0175-7598  |q 99:20<8527  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6615-0  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6615-0  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Wang  |D Junming  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Gao  |D Dongfang  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Yu  |D Xiaoli  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Li  |D Wen  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Qi  |D Qingsheng  |u State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/20(2015-10-01), 8527-8536  |x 0175-7598  |q 99:20<8527  |1 2015  |2 99  |o 253