Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization
To improve the screening efficiency of high-yield neomycin sulfate (NM) <i>Streptomyces fradiae</i> strains after mutagenesis, a high-throughput screening method using streptomycin resistance prescreening (8 μg/mL) and a 24-deep well plates/microplate reader (trypan blue spectrophotometr...
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MDPI AG
2022-01-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/10/1/94 |
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| author | Fei Yu Min Zhang Junfeng Sun Fang Wang Xiangfei Li Yan Liu Zhou Wang Xinrui Zhao Jianghua Li Jian Chen Guocheng Du Zhenglian Xue |
| author_facet | Fei Yu Min Zhang Junfeng Sun Fang Wang Xiangfei Li Yan Liu Zhou Wang Xinrui Zhao Jianghua Li Jian Chen Guocheng Du Zhenglian Xue |
| author_sort | Fei Yu |
| collection | DOAJ |
| description | To improve the screening efficiency of high-yield neomycin sulfate (NM) <i>Streptomyces fradiae</i> strains after mutagenesis, a high-throughput screening method using streptomycin resistance prescreening (8 μg/mL) and a 24-deep well plates/microplate reader (trypan blue spectrophotometry) rescreening strategy was developed. Using this approach, we identified a high-producing NM mutant strain, <i>Sf</i>6-2, via six rounds of atmospheric and room temperature plasma (ARTP) mutagenesis and screening. The mutant displayed a NM potency of 7780 ± 110 U/mL and remarkably stable genetic properties over six generations. Furthermore, the key components (soluble starch, peptone, and (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>) affecting NM potency in fermentation medium were selected using Plackett-Burman and optimized by Box-Behnken designs. Finally, the NM potency of <i>Sf</i>6-2 was increased to 10,849 ± 141 U/mL at the optimal concentration of each factor (73.98 g/L, 9.23 g/L, and 5.99 g/L, respectively), and it exhibited about a 40% and 100% enhancement when compared with before optimization conditions and the wild-type strain, respectively. In this study, we provide a new <i>S. fradiae</i> NM production strategy and generate valuable insights for the breeding and screening of other microorganisms. |
| first_indexed | 2024-03-10T00:54:23Z |
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| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T00:54:23Z |
| publishDate | 2022-01-01 |
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| series | Microorganisms |
| spelling | doaj.art-f9528976e264405d943a75f729f9caec2023-11-23T14:47:14ZengMDPI AGMicroorganisms2076-26072022-01-011019410.3390/microorganisms10010094Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium OptimizationFei Yu0Min Zhang1Junfeng Sun2Fang Wang3Xiangfei Li4Yan Liu5Zhou Wang6Xinrui Zhao7Jianghua Li8Jian Chen9Guocheng Du10Zhenglian Xue11Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, ChinaMicroorganism Fermentation Engineering and Technology Research Center of Anhui Province, College of Biologic & Food Engineering, Anhui Polytechnic University, 8 Middle Beijing Road, Wuhu 241000, ChinaMicroorganism Fermentation Engineering and Technology Research Center of Anhui Province, College of Biologic & Food Engineering, Anhui Polytechnic University, 8 Middle Beijing Road, Wuhu 241000, ChinaMicroorganism Fermentation Engineering and Technology Research Center of Anhui Province, College of Biologic & Food Engineering, Anhui Polytechnic University, 8 Middle Beijing Road, Wuhu 241000, ChinaKey Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, ChinaMicroorganism Fermentation Engineering and Technology Research Center of Anhui Province, College of Biologic & Food Engineering, Anhui Polytechnic University, 8 Middle Beijing Road, Wuhu 241000, ChinaMicroorganism Fermentation Engineering and Technology Research Center of Anhui Province, College of Biologic & Food Engineering, Anhui Polytechnic University, 8 Middle Beijing Road, Wuhu 241000, ChinaKey Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, ChinaMicroorganism Fermentation Engineering and Technology Research Center of Anhui Province, College of Biologic & Food Engineering, Anhui Polytechnic University, 8 Middle Beijing Road, Wuhu 241000, ChinaTo improve the screening efficiency of high-yield neomycin sulfate (NM) <i>Streptomyces fradiae</i> strains after mutagenesis, a high-throughput screening method using streptomycin resistance prescreening (8 μg/mL) and a 24-deep well plates/microplate reader (trypan blue spectrophotometry) rescreening strategy was developed. Using this approach, we identified a high-producing NM mutant strain, <i>Sf</i>6-2, via six rounds of atmospheric and room temperature plasma (ARTP) mutagenesis and screening. The mutant displayed a NM potency of 7780 ± 110 U/mL and remarkably stable genetic properties over six generations. Furthermore, the key components (soluble starch, peptone, and (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>) affecting NM potency in fermentation medium were selected using Plackett-Burman and optimized by Box-Behnken designs. Finally, the NM potency of <i>Sf</i>6-2 was increased to 10,849 ± 141 U/mL at the optimal concentration of each factor (73.98 g/L, 9.23 g/L, and 5.99 g/L, respectively), and it exhibited about a 40% and 100% enhancement when compared with before optimization conditions and the wild-type strain, respectively. In this study, we provide a new <i>S. fradiae</i> NM production strategy and generate valuable insights for the breeding and screening of other microorganisms.https://www.mdpi.com/2076-2607/10/1/94neomycin sulfate (NM)<i>Streptomyces fradiae</i>high-throughput screeningatmospheric and room temperature plasma (ARTP) mutagenesisfermentation medium optimization |
| spellingShingle | Fei Yu Min Zhang Junfeng Sun Fang Wang Xiangfei Li Yan Liu Zhou Wang Xinrui Zhao Jianghua Li Jian Chen Guocheng Du Zhenglian Xue Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization Microorganisms neomycin sulfate (NM) <i>Streptomyces fradiae</i> high-throughput screening atmospheric and room temperature plasma (ARTP) mutagenesis fermentation medium optimization |
| title | Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization |
| title_full | Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization |
| title_fullStr | Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization |
| title_full_unstemmed | Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization |
| title_short | Improved Neomycin Sulfate Potency in <i>Streptomyces fradiae</i> Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization |
| title_sort | improved neomycin sulfate potency in i streptomyces fradiae i using atmospheric and room temperature plasma artp mutagenesis and fermentation medium optimization |
| topic | neomycin sulfate (NM) <i>Streptomyces fradiae</i> high-throughput screening atmospheric and room temperature plasma (ARTP) mutagenesis fermentation medium optimization |
| url | https://www.mdpi.com/2076-2607/10/1/94 |
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