Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i>
The clinical antibiotic gentamicin is a mixture of several difficult-to-separate components, the minor group of which is gentamicin C1a, a precursor for the synthesis of the high-efficacy and low-toxicity antibiotic etimicin. This study aimed to achieve the high production of gentamicin as well as g...
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| Format: | Article |
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MDPI AG
2023-11-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/9/12/997 |
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| author | Ping Yang Huimin Lin Xiaowei Wu Yu Yin Ji’an Li Daijie Chen |
| author_facet | Ping Yang Huimin Lin Xiaowei Wu Yu Yin Ji’an Li Daijie Chen |
| author_sort | Ping Yang |
| collection | DOAJ |
| description | The clinical antibiotic gentamicin is a mixture of several difficult-to-separate components, the minor group of which is gentamicin C1a, a precursor for the synthesis of the high-efficacy and low-toxicity antibiotic etimicin. This study aimed to achieve the high production of gentamicin as well as gentamicin C1a. In this study, the influence of organic and inorganic salts on the gentamicin production was screened and label-free proteomics was used to determine the mechanisms responsible for the effects. In 25 L fermentation experiments, the addition of 0.1% CaCl<sub>2</sub> and 0.3% sodium citrate increased gentamicin titers by 11.5% (2398 μg/mL vs. 2150 μg/mL), while the C1a ratio increased from 38% to 42%. The results showed that CaCl<sub>2</sub> downregulated the synthesis and metabolism of the tetrapyrrole pathway and the GenK protein (0.08-fold) in the gentamicin synthesis pathway, whereas sodium citrate downregulated key proteins in the glycosylation pathway and tricarboxylic acid pathway. Thus, CaCl<sub>2</sub> caused changes in methylation during the synthesis of gentamicin, increasing the proportion of gentamicin C1a. In contrast, sodium citrate inhibited primary metabolism to promote the production of secondary metabolites of gentamicin. This study provided a basis for the co-production of gentamicin C1a mono-component and gentamicin multicomponent. |
| first_indexed | 2024-03-08T20:46:54Z |
| format | Article |
| id | doaj.art-81a506d1579142cba8e46d11d601fe64 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-08T20:46:54Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-81a506d1579142cba8e46d11d601fe642023-12-22T14:07:50ZengMDPI AGFermentation2311-56372023-11-0191299710.3390/fermentation9120997Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i>Ping Yang0Huimin Lin1Xiaowei Wu2Yu Yin3Ji’an Li4Daijie Chen5School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, ChinaState Key Lab. of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, China State Institute of Pharmaceutical Industry, Shanghai 201203, ChinaSchool of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, ChinaSchool of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, ChinaState Key Lab. of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, China State Institute of Pharmaceutical Industry, Shanghai 201203, ChinaSchool of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, ChinaThe clinical antibiotic gentamicin is a mixture of several difficult-to-separate components, the minor group of which is gentamicin C1a, a precursor for the synthesis of the high-efficacy and low-toxicity antibiotic etimicin. This study aimed to achieve the high production of gentamicin as well as gentamicin C1a. In this study, the influence of organic and inorganic salts on the gentamicin production was screened and label-free proteomics was used to determine the mechanisms responsible for the effects. In 25 L fermentation experiments, the addition of 0.1% CaCl<sub>2</sub> and 0.3% sodium citrate increased gentamicin titers by 11.5% (2398 μg/mL vs. 2150 μg/mL), while the C1a ratio increased from 38% to 42%. The results showed that CaCl<sub>2</sub> downregulated the synthesis and metabolism of the tetrapyrrole pathway and the GenK protein (0.08-fold) in the gentamicin synthesis pathway, whereas sodium citrate downregulated key proteins in the glycosylation pathway and tricarboxylic acid pathway. Thus, CaCl<sub>2</sub> caused changes in methylation during the synthesis of gentamicin, increasing the proportion of gentamicin C1a. In contrast, sodium citrate inhibited primary metabolism to promote the production of secondary metabolites of gentamicin. This study provided a basis for the co-production of gentamicin C1a mono-component and gentamicin multicomponent.https://www.mdpi.com/2311-5637/9/12/997gentamicingentamicin C1a<i>Micromonospora echinospora</i>fermentationlabel-free proteomics |
| spellingShingle | Ping Yang Huimin Lin Xiaowei Wu Yu Yin Ji’an Li Daijie Chen Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i> Fermentation gentamicin gentamicin C1a <i>Micromonospora echinospora</i> fermentation label-free proteomics |
| title | Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i> |
| title_full | Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i> |
| title_fullStr | Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i> |
| title_full_unstemmed | Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i> |
| title_short | Proteomic Analysis of the Effect of CaCl<sub>2</sub> and Sodium Citrate on Gentamicin Biosynthesis of <i>Micromonospora echinospora SIPI-GM.01</i> |
| title_sort | proteomic analysis of the effect of cacl sub 2 sub and sodium citrate on gentamicin biosynthesis of i micromonospora echinospora sipi gm 01 i |
| topic | gentamicin gentamicin C1a <i>Micromonospora echinospora</i> fermentation label-free proteomics |
| url | https://www.mdpi.com/2311-5637/9/12/997 |
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