Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulus
ε-Poly-L-lysine (ε-PL) is a natural amino acid polymer produced by microbial fermentation. It has been mainly used as a preservative in the food and cosmetics industries, as a drug carrier in medicines, and as a gene carrier in gene therapy. ε-PL synthase is the key enzyme responsible for the polyme...
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Frontiers Media S.A.
2020-04-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fbioe.2020.00288/full |
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| author | Aixia Wang Wenzhe Tian Lei Cheng Youqiang Xu Xiuwen Wang Jiayang Qin Bo Yu |
| author_facet | Aixia Wang Wenzhe Tian Lei Cheng Youqiang Xu Xiuwen Wang Jiayang Qin Bo Yu |
| author_sort | Aixia Wang |
| collection | DOAJ |
| description | ε-Poly-L-lysine (ε-PL) is a natural amino acid polymer produced by microbial fermentation. It has been mainly used as a preservative in the food and cosmetics industries, as a drug carrier in medicines, and as a gene carrier in gene therapy. ε-PL synthase is the key enzyme responsible for the polymerization of L-lysine to form ε-PL. In this study, the ε-PL synthase gene was overexpressed in Streptomyces albulus CICC 11022 by using the kasOp∗ promoter and the ribosome binding site from the capsid protein of phage ϕC31, which resulted in a genetically engineered strain Q-PL2. The titers of ε-PL produced by Q-PL2 were 88.2% ± 8.3% higher than that produced by the wild strain in shake flask fermentation. With the synergistic effect of 2 g/L sodium citrate, the titers of ε-PL produced by Q-PL2 were 211.2% ± 17.4% higher than that produced by the wild strain. In fed-batch fermentations, 20.1 ± 1.3 g/L of ε-PL was produced by S. albulus Q-PL2 in 72 h with a productivity of 6.7 ± 0.4 g/L/day, which was 3.2 ± 0.3-fold of that produced by the wild strain. These results indicate that ε-PL synthase is one of the rate-limiting enzymes in ε-PL synthesis pathway and lays a foundation for further improving the ε-PL production ability of S. albulus by metabolic engineering. |
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| language | English |
| last_indexed | 2024-12-11T09:46:10Z |
| publishDate | 2020-04-01 |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-d05a3a27290348d18ede6dbf2ffb74b52022-12-22T01:12:34ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-04-01810.3389/fbioe.2020.00288534580Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulusAixia Wang0Wenzhe Tian1Lei Cheng2Youqiang Xu3Xiuwen Wang4Jiayang Qin5Bo Yu6College of Pharmacy, Binzhou Medical University, Yantai, ChinaCollege of Pharmacy, Binzhou Medical University, Yantai, ChinaBeijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University (BTBU), Beijing, ChinaBeijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University (BTBU), Beijing, ChinaCollege of Pharmacy, Binzhou Medical University, Yantai, ChinaCollege of Pharmacy, Binzhou Medical University, Yantai, ChinaCAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, Chinaε-Poly-L-lysine (ε-PL) is a natural amino acid polymer produced by microbial fermentation. It has been mainly used as a preservative in the food and cosmetics industries, as a drug carrier in medicines, and as a gene carrier in gene therapy. ε-PL synthase is the key enzyme responsible for the polymerization of L-lysine to form ε-PL. In this study, the ε-PL synthase gene was overexpressed in Streptomyces albulus CICC 11022 by using the kasOp∗ promoter and the ribosome binding site from the capsid protein of phage ϕC31, which resulted in a genetically engineered strain Q-PL2. The titers of ε-PL produced by Q-PL2 were 88.2% ± 8.3% higher than that produced by the wild strain in shake flask fermentation. With the synergistic effect of 2 g/L sodium citrate, the titers of ε-PL produced by Q-PL2 were 211.2% ± 17.4% higher than that produced by the wild strain. In fed-batch fermentations, 20.1 ± 1.3 g/L of ε-PL was produced by S. albulus Q-PL2 in 72 h with a productivity of 6.7 ± 0.4 g/L/day, which was 3.2 ± 0.3-fold of that produced by the wild strain. These results indicate that ε-PL synthase is one of the rate-limiting enzymes in ε-PL synthesis pathway and lays a foundation for further improving the ε-PL production ability of S. albulus by metabolic engineering.https://www.frontiersin.org/article/10.3389/fbioe.2020.00288/fullStreptomyces albulusε-PL synthaseoverexpressioncitratesynergistic effect |
| spellingShingle | Aixia Wang Wenzhe Tian Lei Cheng Youqiang Xu Xiuwen Wang Jiayang Qin Bo Yu Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulus Frontiers in Bioengineering and Biotechnology Streptomyces albulus ε-PL synthase overexpression citrate synergistic effect |
| title | Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulus |
| title_full | Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulus |
| title_fullStr | Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulus |
| title_full_unstemmed | Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulus |
| title_short | Enhanced ε-Poly-L-Lysine Production by the Synergistic Effect of ε-Poly-L-Lysine Synthetase Overexpression and Citrate in Streptomyces albulus |
| title_sort | enhanced ε poly l lysine production by the synergistic effect of ε poly l lysine synthetase overexpression and citrate in streptomyces albulus |
| topic | Streptomyces albulus ε-PL synthase overexpression citrate synergistic effect |
| url | https://www.frontiersin.org/article/10.3389/fbioe.2020.00288/full |
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