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Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene

Poly-gamma-glutamic acid (PGA) is a promising bio-based polymer that shares many functions with poly (acrylic acid) and its derivatives. Thus, technologies for efficient production and molecular size control of PGA are required to expand the application of this useful biopolymer. In Bacillus strains...

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Autores principales: Sawada, Kazuhisa, Araki, Hiroyuki, Takimura, Yasushi, Masuda, Kenta, Kageyama, Yasushi, Ozaki, Katsuya, Hagihara, Hiroshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6029982/
https://www.ncbi.nlm.nih.gov/pubmed/29971620
http://dx.doi.org/10.1186/s13568-018-0636-x
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author Sawada, Kazuhisa
Araki, Hiroyuki
Takimura, Yasushi
Masuda, Kenta
Kageyama, Yasushi
Ozaki, Katsuya
Hagihara, Hiroshi
author_facet Sawada, Kazuhisa
Araki, Hiroyuki
Takimura, Yasushi
Masuda, Kenta
Kageyama, Yasushi
Ozaki, Katsuya
Hagihara, Hiroshi
author_sort Sawada, Kazuhisa
collection PubMed
description Poly-gamma-glutamic acid (PGA) is a promising bio-based polymer that shares many functions with poly (acrylic acid) and its derivatives. Thus, technologies for efficient production and molecular size control of PGA are required to expand the application of this useful biopolymer. In Bacillus strains, PGA is synthesized by the PgsBCA protein complex, which is encoded by the pgsBCA gene operon, otherwise is known as ywsC and ywtAB operons and/or capBCA operon. Hence, we investigated responsible components of the PgsBCA complex in B. subtilis for over-production of PGA. In particular, we constructed genomic pgsBCA gene-deletion mutants of B. subtilis. And also, we assembled high copy-number plasmids harboring σA-dependent promoter, leading to high-level expression of all combinations of pgsBCA, pgsBC, pgsBA, pgsCA, pgsB, pgsC, and/or pgsA genes. Subsequently, PGA production of the transformed B. subtilis mutant was determined in batch fermentation using medium supplemented with l-glutamate. PGA production by the transformants introduced with pgsBC genes (lacking the genomic pgsBCA genes) was 26.0 ± 3.0 g L(−1), and the enantiomeric ratio of d- and l-glutamic acid (d/l-ratio) in the produced PGA was 5/95. In contrast, d/l-ratio of produced PGA by the transformants introduced with pgsBCA genes (control strains) was 75/25. In conclusion, B. subtilis without pgsA gene could over-produce PGA with an l-rich enantiomeric ratio.
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spelling pubmed-60299822018-07-23 Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene Sawada, Kazuhisa Araki, Hiroyuki Takimura, Yasushi Masuda, Kenta Kageyama, Yasushi Ozaki, Katsuya Hagihara, Hiroshi AMB Express Original Article Poly-gamma-glutamic acid (PGA) is a promising bio-based polymer that shares many functions with poly (acrylic acid) and its derivatives. Thus, technologies for efficient production and molecular size control of PGA are required to expand the application of this useful biopolymer. In Bacillus strains, PGA is synthesized by the PgsBCA protein complex, which is encoded by the pgsBCA gene operon, otherwise is known as ywsC and ywtAB operons and/or capBCA operon. Hence, we investigated responsible components of the PgsBCA complex in B. subtilis for over-production of PGA. In particular, we constructed genomic pgsBCA gene-deletion mutants of B. subtilis. And also, we assembled high copy-number plasmids harboring σA-dependent promoter, leading to high-level expression of all combinations of pgsBCA, pgsBC, pgsBA, pgsCA, pgsB, pgsC, and/or pgsA genes. Subsequently, PGA production of the transformed B. subtilis mutant was determined in batch fermentation using medium supplemented with l-glutamate. PGA production by the transformants introduced with pgsBC genes (lacking the genomic pgsBCA genes) was 26.0 ± 3.0 g L(−1), and the enantiomeric ratio of d- and l-glutamic acid (d/l-ratio) in the produced PGA was 5/95. In contrast, d/l-ratio of produced PGA by the transformants introduced with pgsBCA genes (control strains) was 75/25. In conclusion, B. subtilis without pgsA gene could over-produce PGA with an l-rich enantiomeric ratio. Springer Berlin Heidelberg 2018-07-03 /pmc/articles/PMC6029982/ /pubmed/29971620 http://dx.doi.org/10.1186/s13568-018-0636-x Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Sawada, Kazuhisa
Araki, Hiroyuki
Takimura, Yasushi
Masuda, Kenta
Kageyama, Yasushi
Ozaki, Katsuya
Hagihara, Hiroshi
Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene
title Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene
title_full Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene
title_fullStr Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene
title_full_unstemmed Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene
title_short Poly-l-gamma-glutamic acid production by recombinant Bacillus subtilis without pgsA gene
title_sort poly-l-gamma-glutamic acid production by recombinant bacillus subtilis without pgsa gene
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6029982/
https://www.ncbi.nlm.nih.gov/pubmed/29971620
http://dx.doi.org/10.1186/s13568-018-0636-x
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