Cargando…
Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS
In pharmaceutical industry, isepamicin is mainly manufactured from gentamicin B, which is produced by Micromonospora echinospora as a minor component of the gentamicin complex. Improvement of gentamicin B production through metabolic engineering is therefore important to satisfy the increasing deman...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6838515/ https://www.ncbi.nlm.nih.gov/pubmed/31720212 http://dx.doi.org/10.1016/j.mec.2019.e00096 |
_version_ | 1783467236777787392 |
---|---|
author | Chang, Yingying Chai, Baozhong Ding, Yunkun He, Min Zheng, Linghui Teng, Yun Deng, Zixin Yu, Yi Liu, Tiangang |
author_facet | Chang, Yingying Chai, Baozhong Ding, Yunkun He, Min Zheng, Linghui Teng, Yun Deng, Zixin Yu, Yi Liu, Tiangang |
author_sort | Chang, Yingying |
collection | PubMed |
description | In pharmaceutical industry, isepamicin is mainly manufactured from gentamicin B, which is produced by Micromonospora echinospora as a minor component of the gentamicin complex. Improvement of gentamicin B production through metabolic engineering is therefore important to satisfy the increasing demand for isepamicin. We hypothesized that gentamicin B was generated from gentamicin JI-20A via deamination of the C2’ amino group. Using kanJ and kanK as the gene probes, we identified the putative deamination-related genes, genR and genS, through genome mining of the gentamicin B producing strain M. echinospora CCTCC M 2018898. Interestingly, genR and genS constitute a gene cassette located approximately 28.7 kb away from the gentamicin gene cluster. Gene knockout of genR and genS almost abolished the production of gentamicin B in the mutant strain, suggesting that these two genes, which are responsible for the last steps in gentamicin B biosynthesis, constitute the missing part of the known gentamicin biosynthetic pathway. Based on these finding, we successfully constructed a gentamicin B high-yielding strain (798 mg/L), in which an overexpression cassette of genR and genS was introduced. Our work fills the missing piece to solve the puzzle of gentamicin B biosynthesis and may inspire future metabolic engineering efforts to generate gentamycin B high-yielding strains that could eventually satisfy the need for industrial manufacturing of isepamicin. |
format | Online Article Text |
id | pubmed-6838515 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-68385152019-11-12 Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS Chang, Yingying Chai, Baozhong Ding, Yunkun He, Min Zheng, Linghui Teng, Yun Deng, Zixin Yu, Yi Liu, Tiangang Metab Eng Commun Article In pharmaceutical industry, isepamicin is mainly manufactured from gentamicin B, which is produced by Micromonospora echinospora as a minor component of the gentamicin complex. Improvement of gentamicin B production through metabolic engineering is therefore important to satisfy the increasing demand for isepamicin. We hypothesized that gentamicin B was generated from gentamicin JI-20A via deamination of the C2’ amino group. Using kanJ and kanK as the gene probes, we identified the putative deamination-related genes, genR and genS, through genome mining of the gentamicin B producing strain M. echinospora CCTCC M 2018898. Interestingly, genR and genS constitute a gene cassette located approximately 28.7 kb away from the gentamicin gene cluster. Gene knockout of genR and genS almost abolished the production of gentamicin B in the mutant strain, suggesting that these two genes, which are responsible for the last steps in gentamicin B biosynthesis, constitute the missing part of the known gentamicin biosynthetic pathway. Based on these finding, we successfully constructed a gentamicin B high-yielding strain (798 mg/L), in which an overexpression cassette of genR and genS was introduced. Our work fills the missing piece to solve the puzzle of gentamicin B biosynthesis and may inspire future metabolic engineering efforts to generate gentamycin B high-yielding strains that could eventually satisfy the need for industrial manufacturing of isepamicin. Elsevier 2019-07-24 /pmc/articles/PMC6838515/ /pubmed/31720212 http://dx.doi.org/10.1016/j.mec.2019.e00096 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Chang, Yingying Chai, Baozhong Ding, Yunkun He, Min Zheng, Linghui Teng, Yun Deng, Zixin Yu, Yi Liu, Tiangang Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS |
title | Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS |
title_full | Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS |
title_fullStr | Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS |
title_full_unstemmed | Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS |
title_short | Overproduction of gentamicin B in industrial strain Micromonospora echinospora CCTCC M 2018898 by cloning of the missing genes genR and genS |
title_sort | overproduction of gentamicin b in industrial strain micromonospora echinospora cctcc m 2018898 by cloning of the missing genes genr and gens |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6838515/ https://www.ncbi.nlm.nih.gov/pubmed/31720212 http://dx.doi.org/10.1016/j.mec.2019.e00096 |
work_keys_str_mv | AT changyingying overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT chaibaozhong overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT dingyunkun overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT hemin overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT zhenglinghui overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT tengyun overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT dengzixin overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT yuyi overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens AT liutiangang overproductionofgentamicinbinindustrialstrainmicromonosporaechinosporacctccm2018898bycloningofthemissinggenesgenrandgens |