Cargando…
Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21
BACKGROUND: Polyhydroxybutyrate (PHB) is currently the most common polymer produced by natural bacteria and alternative to conventional petrochemical-based plastics due to its similar material properties and biodegradability. Massilia sp. UMI-21, a newly found bacterium, could produce PHB from starc...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10347839/ https://www.ncbi.nlm.nih.gov/pubmed/37452345 http://dx.doi.org/10.1186/s12934-023-02142-x |
_version_ | 1785073610476486656 |
---|---|
author | Jiang, Nan Wang, Ming Song, Linxin Yu, Dengbin Zhou, Shuangzi Li, Yu Li, Haiyan Han, Xuerong |
author_facet | Jiang, Nan Wang, Ming Song, Linxin Yu, Dengbin Zhou, Shuangzi Li, Yu Li, Haiyan Han, Xuerong |
author_sort | Jiang, Nan |
collection | PubMed |
description | BACKGROUND: Polyhydroxybutyrate (PHB) is currently the most common polymer produced by natural bacteria and alternative to conventional petrochemical-based plastics due to its similar material properties and biodegradability. Massilia sp. UMI-21, a newly found bacterium, could produce PHB from starch, maltotriose, or maltose, etc. and could serve as a candidate for seaweed-degrading bioplastic producers. However, the genes involved in PHB metabolism in Massilia sp. UMI-21 are still unclear. RESULTS: In the present study, we assembled and annotated the genome of Massilia sp. UMI-21, identified genes related to the metabolism of PHB, and successfully constructed recombinant Escherichia coli harboring PHB-related genes (phaA2, phaB1 and phaC1) of Massilia sp. UMI-21, which showed up to 139.41% more product. Also, the vgb gene (encoding Vitreoscilla hemoglobin) was introduced into the genetically engineered E. coli and gained up to 117.42% more cell dry weight, 213.30% more PHB-like production and 44.09% more product content. Fermentation products extracted from recombinant E. coli harboring pETDuet1-phaA2phaB1-phaC1 and pETDuet1-phaA2phaB1-phaC1-vgb were identified as PHB by Fourier Transform Infrared and Proton nuclear magnetic resonance spectroscopy analysis. Furthermore, the decomposition temperature at 10% weight loss of PHB extracted from Massilia sp. UMI-21, recombinant E. coli DH5α-pETDuet1-phaA2phaB1-phaC1 and DH5α-pETDuet1-phaA2phaB1-phaC1-vgb was 276.5, 278.7 and 286.3 °C, respectively, showing good thermal stability. CONCLUSIONS: Herein, we presented the whole genome information of PHB-producing Massilia sp. UMI-21 and constructed novel recombinant strains using key genes in PHB synthesis of strain UMI-21 and the vgb gene. This genetically engineered E. coli strain can serve as an effective novel candidate in E. coli cell factory for PHB production by the rapid cell growth and high PHB production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-023-02142-x. |
format | Online Article Text |
id | pubmed-10347839 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-103478392023-07-15 Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21 Jiang, Nan Wang, Ming Song, Linxin Yu, Dengbin Zhou, Shuangzi Li, Yu Li, Haiyan Han, Xuerong Microb Cell Fact Research BACKGROUND: Polyhydroxybutyrate (PHB) is currently the most common polymer produced by natural bacteria and alternative to conventional petrochemical-based plastics due to its similar material properties and biodegradability. Massilia sp. UMI-21, a newly found bacterium, could produce PHB from starch, maltotriose, or maltose, etc. and could serve as a candidate for seaweed-degrading bioplastic producers. However, the genes involved in PHB metabolism in Massilia sp. UMI-21 are still unclear. RESULTS: In the present study, we assembled and annotated the genome of Massilia sp. UMI-21, identified genes related to the metabolism of PHB, and successfully constructed recombinant Escherichia coli harboring PHB-related genes (phaA2, phaB1 and phaC1) of Massilia sp. UMI-21, which showed up to 139.41% more product. Also, the vgb gene (encoding Vitreoscilla hemoglobin) was introduced into the genetically engineered E. coli and gained up to 117.42% more cell dry weight, 213.30% more PHB-like production and 44.09% more product content. Fermentation products extracted from recombinant E. coli harboring pETDuet1-phaA2phaB1-phaC1 and pETDuet1-phaA2phaB1-phaC1-vgb were identified as PHB by Fourier Transform Infrared and Proton nuclear magnetic resonance spectroscopy analysis. Furthermore, the decomposition temperature at 10% weight loss of PHB extracted from Massilia sp. UMI-21, recombinant E. coli DH5α-pETDuet1-phaA2phaB1-phaC1 and DH5α-pETDuet1-phaA2phaB1-phaC1-vgb was 276.5, 278.7 and 286.3 °C, respectively, showing good thermal stability. CONCLUSIONS: Herein, we presented the whole genome information of PHB-producing Massilia sp. UMI-21 and constructed novel recombinant strains using key genes in PHB synthesis of strain UMI-21 and the vgb gene. This genetically engineered E. coli strain can serve as an effective novel candidate in E. coli cell factory for PHB production by the rapid cell growth and high PHB production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-023-02142-x. BioMed Central 2023-07-14 /pmc/articles/PMC10347839/ /pubmed/37452345 http://dx.doi.org/10.1186/s12934-023-02142-x Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Jiang, Nan Wang, Ming Song, Linxin Yu, Dengbin Zhou, Shuangzi Li, Yu Li, Haiyan Han, Xuerong Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21 |
title | Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21 |
title_full | Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21 |
title_fullStr | Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21 |
title_full_unstemmed | Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21 |
title_short | Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21 |
title_sort | polyhydroxybutyrate production by recombinant escherichia coli based on genes related to synthesis pathway of phb from massilia sp. umi-21 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10347839/ https://www.ncbi.nlm.nih.gov/pubmed/37452345 http://dx.doi.org/10.1186/s12934-023-02142-x |
work_keys_str_mv | AT jiangnan polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 AT wangming polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 AT songlinxin polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 AT yudengbin polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 AT zhoushuangzi polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 AT liyu polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 AT lihaiyan polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 AT hanxuerong polyhydroxybutyrateproductionbyrecombinantescherichiacolibasedongenesrelatedtosynthesispathwayofphbfrommassiliaspumi21 |