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Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection

The B12-producing strains Pseudomonas nitroreducens DSM 1650 and Pseudomonas sp. CCUG 2519 (both formerly Pseudomonas denitrificans), with the most distributed pathway among bacteria for exogenous choline/betaine utilization, are promising recombinant hosts for the endogenous production of B12 precu...

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Autores principales: Balabanova, Larissa, Pentekhina, Iuliia, Nedashkovskaya, Olga, Degtyarenko, Anton, Grigorchuk, Valeria, Yugay, Yulia, Vasyutkina, Elena, Kudinova, Olesya, Seitkalieva, Aleksandra, Slepchenko, Lubov, Son, Oksana, Tekutyeva, Liudmila, Shkryl, Yury
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9699165/
https://www.ncbi.nlm.nih.gov/pubmed/36430408
http://dx.doi.org/10.3390/ijms232213934
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author Balabanova, Larissa
Pentekhina, Iuliia
Nedashkovskaya, Olga
Degtyarenko, Anton
Grigorchuk, Valeria
Yugay, Yulia
Vasyutkina, Elena
Kudinova, Olesya
Seitkalieva, Aleksandra
Slepchenko, Lubov
Son, Oksana
Tekutyeva, Liudmila
Shkryl, Yury
author_facet Balabanova, Larissa
Pentekhina, Iuliia
Nedashkovskaya, Olga
Degtyarenko, Anton
Grigorchuk, Valeria
Yugay, Yulia
Vasyutkina, Elena
Kudinova, Olesya
Seitkalieva, Aleksandra
Slepchenko, Lubov
Son, Oksana
Tekutyeva, Liudmila
Shkryl, Yury
author_sort Balabanova, Larissa
collection PubMed
description The B12-producing strains Pseudomonas nitroreducens DSM 1650 and Pseudomonas sp. CCUG 2519 (both formerly Pseudomonas denitrificans), with the most distributed pathway among bacteria for exogenous choline/betaine utilization, are promising recombinant hosts for the endogenous production of B12 precursor betaine by direct methylation of bioavailable glycine or non-proteinogenic β-alanine. Two plasmid-based de novo betaine pathways, distinguished by their enzymes, have provided an expression of the genes encoding for N-methyltransferases of the halotolerant cyanobacterium Aphanothece halophytica or plant Limonium latifolium to synthesize the internal glycine betaine or β-alanine betaine, respectively. These betaines equally allowed the recombinant pseudomonads to grow effectively and to synthesize a high level of cobalamin, as well as to increase their protective properties against abiotic stresses to a degree comparable with the supplementation of an exogenous betaine. Both de novo betaine pathways significantly enforced the protection of bacterial cells against lowering temperature to 15 °C and increasing salinity to 400 mM of NaCl. However, the expression of the single plant-derived gene for the β-alanine-specific N-methyltransferase additionally increased the effectiveness of exogenous glycine betaine almost twofold on cobalamin biosynthesis, probably due to the Pseudomonas’ ability to use two independent pathways, their own choline/betaine pathway and the plant β-alanine betaine biosynthetic pathway.
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spelling pubmed-96991652022-11-26 Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection Balabanova, Larissa Pentekhina, Iuliia Nedashkovskaya, Olga Degtyarenko, Anton Grigorchuk, Valeria Yugay, Yulia Vasyutkina, Elena Kudinova, Olesya Seitkalieva, Aleksandra Slepchenko, Lubov Son, Oksana Tekutyeva, Liudmila Shkryl, Yury Int J Mol Sci Brief Report The B12-producing strains Pseudomonas nitroreducens DSM 1650 and Pseudomonas sp. CCUG 2519 (both formerly Pseudomonas denitrificans), with the most distributed pathway among bacteria for exogenous choline/betaine utilization, are promising recombinant hosts for the endogenous production of B12 precursor betaine by direct methylation of bioavailable glycine or non-proteinogenic β-alanine. Two plasmid-based de novo betaine pathways, distinguished by their enzymes, have provided an expression of the genes encoding for N-methyltransferases of the halotolerant cyanobacterium Aphanothece halophytica or plant Limonium latifolium to synthesize the internal glycine betaine or β-alanine betaine, respectively. These betaines equally allowed the recombinant pseudomonads to grow effectively and to synthesize a high level of cobalamin, as well as to increase their protective properties against abiotic stresses to a degree comparable with the supplementation of an exogenous betaine. Both de novo betaine pathways significantly enforced the protection of bacterial cells against lowering temperature to 15 °C and increasing salinity to 400 mM of NaCl. However, the expression of the single plant-derived gene for the β-alanine-specific N-methyltransferase additionally increased the effectiveness of exogenous glycine betaine almost twofold on cobalamin biosynthesis, probably due to the Pseudomonas’ ability to use two independent pathways, their own choline/betaine pathway and the plant β-alanine betaine biosynthetic pathway. MDPI 2022-11-11 /pmc/articles/PMC9699165/ /pubmed/36430408 http://dx.doi.org/10.3390/ijms232213934 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Brief Report
Balabanova, Larissa
Pentekhina, Iuliia
Nedashkovskaya, Olga
Degtyarenko, Anton
Grigorchuk, Valeria
Yugay, Yulia
Vasyutkina, Elena
Kudinova, Olesya
Seitkalieva, Aleksandra
Slepchenko, Lubov
Son, Oksana
Tekutyeva, Liudmila
Shkryl, Yury
Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection
title Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection
title_full Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection
title_fullStr Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection
title_full_unstemmed Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection
title_short Shift of Choline/Betaine Pathway in Recombinant Pseudomonas for Cobalamin Biosynthesis and Abiotic Stress Protection
title_sort shift of choline/betaine pathway in recombinant pseudomonas for cobalamin biosynthesis and abiotic stress protection
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9699165/
https://www.ncbi.nlm.nih.gov/pubmed/36430408
http://dx.doi.org/10.3390/ijms232213934
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