Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>

ABSTRACT Bacillus methanolicus MGA3 is a thermotolerant and relatively fast-growing methylotroph able to secrete large quantities of glutamate and lysine. These natural characteristics make B. methanolicus a good candidate to become a new industrial chassis organism, especially in a methanol-based e...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Baudoin Delépine, Marina Gil López, Marc Carnicer, Cláudia M. Vicente, Volker F. Wendisch, Stéphanie Heux
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	natural methylotrophy 13C metabolic flux analysis nonstationary MFA Bacillus methanolicus MGA3 methanol Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7d2c6ae488da47398ab71fc9e41bb96c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:7d2c6ae488da47398ab71fc9e41bb96c
record_format	dspace
spelling	oai:doaj.org-article:7d2c6ae488da47398ab71fc9e41bb96c2021-12-02T18:15:46ZCharting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>10.1128/mSystems.00745-202379-5077https://doaj.org/article/7d2c6ae488da47398ab71fc9e41bb96c2020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00745-20https://doaj.org/toc/2379-5077ABSTRACT Bacillus methanolicus MGA3 is a thermotolerant and relatively fast-growing methylotroph able to secrete large quantities of glutamate and lysine. These natural characteristics make B. methanolicus a good candidate to become a new industrial chassis organism, especially in a methanol-based economy. Intriguingly, the only substrates known to support B. methanolicus growth as sole sources of carbon and energy are methanol, mannitol, and, to a lesser extent, glucose and arabitol. Because fluxomics provides the most direct readout of the cellular phenotype, we hypothesized that comparing methylotrophic and nonmethylotrophic metabolic states at the flux level would yield new insights into MGA3 metabolism. In this study, we designed and performed a 13C metabolic flux analysis (13C-MFA) of the facultative methylotroph B. methanolicus MGA3 growing on methanol, mannitol, and arabitol to compare the associated metabolic states. On methanol, results showed a greater flux in the ribulose monophosphate (RuMP) pathway than in the tricarboxylic acid (TCA) cycle, thus validating previous findings on the methylotrophy of B. methanolicus. New insights related to the utilization of cyclic RuMP versus linear dissimilation pathways and between the RuMP variants were generated. Importantly, we demonstrated that the linear detoxification pathways and the malic enzyme shared with the pentose phosphate pathway have an important role in cofactor regeneration. Finally, we identified, for the first time, the metabolic pathway used to assimilate arabitol. Overall, those data provide a better understanding of this strain under various environmental conditions. IMPORTANCE Methanol is inexpensive, is easy to transport, and can be produced both from renewable and from fossil resources without mobilizing arable lands. As such, it is regarded as a potential carbon source to transition toward a greener industrial chemistry. Metabolic engineering of bacteria and yeast able to efficiently consume methanol is expected to provide cell factories that will transform methanol into higher-value chemicals in the so-called methanol economy. Toward that goal, the study of natural methylotrophs such as Bacillus methanolicus is critical to understand the origin of their efficient methylotrophy. This knowledge will then be leveraged to transform such natural strains into new cell factories or to design methylotrophic capability in other strains already used by the industry.Baudoin DelépineMarina Gil LópezMarc CarnicerCláudia M. VicenteVolker F. WendischStéphanie HeuxAmerican Society for Microbiologyarticlenatural methylotrophy13C metabolic flux analysisnonstationary MFABacillus methanolicus MGA3methanolMicrobiologyQR1-502ENmSystems, Vol 5, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	natural methylotrophy 13C metabolic flux analysis nonstationary MFA Bacillus methanolicus MGA3 methanol Microbiology QR1-502
spellingShingle	natural methylotrophy 13C metabolic flux analysis nonstationary MFA Bacillus methanolicus MGA3 methanol Microbiology QR1-502 Baudoin Delépine Marina Gil López Marc Carnicer Cláudia M. Vicente Volker F. Wendisch Stéphanie Heux Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>
description	ABSTRACT Bacillus methanolicus MGA3 is a thermotolerant and relatively fast-growing methylotroph able to secrete large quantities of glutamate and lysine. These natural characteristics make B. methanolicus a good candidate to become a new industrial chassis organism, especially in a methanol-based economy. Intriguingly, the only substrates known to support B. methanolicus growth as sole sources of carbon and energy are methanol, mannitol, and, to a lesser extent, glucose and arabitol. Because fluxomics provides the most direct readout of the cellular phenotype, we hypothesized that comparing methylotrophic and nonmethylotrophic metabolic states at the flux level would yield new insights into MGA3 metabolism. In this study, we designed and performed a 13C metabolic flux analysis (13C-MFA) of the facultative methylotroph B. methanolicus MGA3 growing on methanol, mannitol, and arabitol to compare the associated metabolic states. On methanol, results showed a greater flux in the ribulose monophosphate (RuMP) pathway than in the tricarboxylic acid (TCA) cycle, thus validating previous findings on the methylotrophy of B. methanolicus. New insights related to the utilization of cyclic RuMP versus linear dissimilation pathways and between the RuMP variants were generated. Importantly, we demonstrated that the linear detoxification pathways and the malic enzyme shared with the pentose phosphate pathway have an important role in cofactor regeneration. Finally, we identified, for the first time, the metabolic pathway used to assimilate arabitol. Overall, those data provide a better understanding of this strain under various environmental conditions. IMPORTANCE Methanol is inexpensive, is easy to transport, and can be produced both from renewable and from fossil resources without mobilizing arable lands. As such, it is regarded as a potential carbon source to transition toward a greener industrial chemistry. Metabolic engineering of bacteria and yeast able to efficiently consume methanol is expected to provide cell factories that will transform methanol into higher-value chemicals in the so-called methanol economy. Toward that goal, the study of natural methylotrophs such as Bacillus methanolicus is critical to understand the origin of their efficient methylotrophy. This knowledge will then be leveraged to transform such natural strains into new cell factories or to design methylotrophic capability in other strains already used by the industry.
format	article
author	Baudoin Delépine Marina Gil López Marc Carnicer Cláudia M. Vicente Volker F. Wendisch Stéphanie Heux
author_facet	Baudoin Delépine Marina Gil López Marc Carnicer Cláudia M. Vicente Volker F. Wendisch Stéphanie Heux
author_sort	Baudoin Delépine
title	Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>
title_short	Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>
title_full	Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>
title_fullStr	Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>
title_full_unstemmed	Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>
title_sort	charting the metabolic landscape of the facultative methylotroph <named-content content-type="genus-species">bacillus methanolicus</named-content>
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/7d2c6ae488da47398ab71fc9e41bb96c
work_keys_str_mv	AT baudoindelepine chartingthemetaboliclandscapeofthefacultativemethylotrophnamedcontentcontenttypegenusspeciesbacillusmethanolicusnamedcontent AT marinagillopez chartingthemetaboliclandscapeofthefacultativemethylotrophnamedcontentcontenttypegenusspeciesbacillusmethanolicusnamedcontent AT marccarnicer chartingthemetaboliclandscapeofthefacultativemethylotrophnamedcontentcontenttypegenusspeciesbacillusmethanolicusnamedcontent AT claudiamvicente chartingthemetaboliclandscapeofthefacultativemethylotrophnamedcontentcontenttypegenusspeciesbacillusmethanolicusnamedcontent AT volkerfwendisch chartingthemetaboliclandscapeofthefacultativemethylotrophnamedcontentcontenttypegenusspeciesbacillusmethanolicusnamedcontent AT stephanieheux chartingthemetaboliclandscapeofthefacultativemethylotrophnamedcontentcontenttypegenusspeciesbacillusmethanolicusnamedcontent
_version_	1718378342743277568

Charting the Metabolic Landscape of the Facultative Methylotroph <named-content content-type="genus-species">Bacillus methanolicus</named-content>

Ejemplares similares