Cargando…
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...
Autores principales: | , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1784839004323053568 |
---|---|
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. |
format | Online Article Text |
id | pubmed-9699165 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT balabanovalarissa shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT pentekhinaiuliia shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT nedashkovskayaolga shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT degtyarenkoanton shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT grigorchukvaleria shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT yugayyulia shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT vasyutkinaelena shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT kudinovaolesya shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT seitkalievaaleksandra shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT slepchenkolubov shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT sonoksana shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT tekutyevaliudmila shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection AT shkrylyury shiftofcholinebetainepathwayinrecombinantpseudomonasforcobalaminbiosynthesisandabioticstressprotection |