Evolution of a chimeric aspartate kinase for L-lysine production using a synthetic RNA device
Gespeichert in:
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)
Online Zugang:
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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 | ||