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Development of Bacillus subtilis mutants to produce tryptophan in pigs

OBJECTIVES: To generate tryptophan-overproducing Bacillus subtilis strains for in situ use in pigs, to reduce the feed cost for farmers and nitrogen pollution. RESULTS: A novel concept has been investigated—to generate B. subtilis strains able to produce tryptophan (Trp) in situ in pigs. Mutagenesis...

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Autores principales: Bjerre, Karin, Cantor, Mette D., Nørgaard, Jan V., Poulsen, Hanne D., Blaabjerg, Karoline, Canibe, Nuria, Jensen, Bent B., Stuer-Lauridsen, Birgitte, Nielsen, Bea, Derkx, Patrick M. F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5247549/
https://www.ncbi.nlm.nih.gov/pubmed/27812824
http://dx.doi.org/10.1007/s10529-016-2245-6
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author Bjerre, Karin
Cantor, Mette D.
Nørgaard, Jan V.
Poulsen, Hanne D.
Blaabjerg, Karoline
Canibe, Nuria
Jensen, Bent B.
Stuer-Lauridsen, Birgitte
Nielsen, Bea
Derkx, Patrick M. F.
author_facet Bjerre, Karin
Cantor, Mette D.
Nørgaard, Jan V.
Poulsen, Hanne D.
Blaabjerg, Karoline
Canibe, Nuria
Jensen, Bent B.
Stuer-Lauridsen, Birgitte
Nielsen, Bea
Derkx, Patrick M. F.
author_sort Bjerre, Karin
collection PubMed
description OBJECTIVES: To generate tryptophan-overproducing Bacillus subtilis strains for in situ use in pigs, to reduce the feed cost for farmers and nitrogen pollution. RESULTS: A novel concept has been investigated—to generate B. subtilis strains able to produce tryptophan (Trp) in situ in pigs. Mutagenesis by UV was combined with selection on Trp and purine analogues in an iterative process. Two mutants from different wild types were obtained, mutant 1 (M1) produced 1 mg Trp/l and mutant 2 (M2) 14 mg Trp/l. Genome sequence analysis revealed that M1 had three single nuclear polymorphisms (SNPs) and M2 had two SNPs compared to the wild type strains. In both mutants SNPs were found in genes regulating tryptophan synthesis. Reverse transcription PCR confirmed up-regulation of the tryptophan synthesis genes in both mutants, the expression was up to 3 times higher in M2 than in M1. CONCLUSIONS: Tryptophan-excreting B. subtilis strains were obtained with UV-mutagenesis and analogue selection and can be used in animal feed applications. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10529-016-2245-6) contains supplementary material, which is available to authorized users.
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spelling pubmed-52475492017-02-01 Development of Bacillus subtilis mutants to produce tryptophan in pigs Bjerre, Karin Cantor, Mette D. Nørgaard, Jan V. Poulsen, Hanne D. Blaabjerg, Karoline Canibe, Nuria Jensen, Bent B. Stuer-Lauridsen, Birgitte Nielsen, Bea Derkx, Patrick M. F. Biotechnol Lett Original Research Paper OBJECTIVES: To generate tryptophan-overproducing Bacillus subtilis strains for in situ use in pigs, to reduce the feed cost for farmers and nitrogen pollution. RESULTS: A novel concept has been investigated—to generate B. subtilis strains able to produce tryptophan (Trp) in situ in pigs. Mutagenesis by UV was combined with selection on Trp and purine analogues in an iterative process. Two mutants from different wild types were obtained, mutant 1 (M1) produced 1 mg Trp/l and mutant 2 (M2) 14 mg Trp/l. Genome sequence analysis revealed that M1 had three single nuclear polymorphisms (SNPs) and M2 had two SNPs compared to the wild type strains. In both mutants SNPs were found in genes regulating tryptophan synthesis. Reverse transcription PCR confirmed up-regulation of the tryptophan synthesis genes in both mutants, the expression was up to 3 times higher in M2 than in M1. CONCLUSIONS: Tryptophan-excreting B. subtilis strains were obtained with UV-mutagenesis and analogue selection and can be used in animal feed applications. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10529-016-2245-6) contains supplementary material, which is available to authorized users. Springer Netherlands 2016-11-03 2017 /pmc/articles/PMC5247549/ /pubmed/27812824 http://dx.doi.org/10.1007/s10529-016-2245-6 Text en © The Author(s) 2016 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 Research Paper
Bjerre, Karin
Cantor, Mette D.
Nørgaard, Jan V.
Poulsen, Hanne D.
Blaabjerg, Karoline
Canibe, Nuria
Jensen, Bent B.
Stuer-Lauridsen, Birgitte
Nielsen, Bea
Derkx, Patrick M. F.
Development of Bacillus subtilis mutants to produce tryptophan in pigs
title Development of Bacillus subtilis mutants to produce tryptophan in pigs
title_full Development of Bacillus subtilis mutants to produce tryptophan in pigs
title_fullStr Development of Bacillus subtilis mutants to produce tryptophan in pigs
title_full_unstemmed Development of Bacillus subtilis mutants to produce tryptophan in pigs
title_short Development of Bacillus subtilis mutants to produce tryptophan in pigs
title_sort development of bacillus subtilis mutants to produce tryptophan in pigs
topic Original Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5247549/
https://www.ncbi.nlm.nih.gov/pubmed/27812824
http://dx.doi.org/10.1007/s10529-016-2245-6
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