TY - JOUR
T1 - Charting the metabolic landscape of the facultative methylotroph bacillus methanolicus
AU - Delépine, Baudoin
AU - López, Marina Gil
AU - Carnicer, Marc
AU - Vicente, Cláudia M.
AU - Wendisch, Volker F.
AU - Heux, Stéphanie
N1 - Funding Information:
We thank MetaToul (www.metatoul.fr), which is part of MetaboHub (ANR-11-INBS-0010, www.metabohub.fr), for their help in collecting, processing, and interpreting 13C NMR and MS data. We also thank Marcus Persicke, Maud Heuillet, Edern Cahoreau, Lindsay Peyriga, Pierre Millard, Gilles Vieira, and Jean-Charles Portais for the insights they provided. This work was supported by ERA-CoBioTech's project C1Pro (ANR-17-COBI-0003-05 and FNR-22023617).
Funding Information:
We thank MetaToul (www.metatoul.fr), which is part of MetaboHub (ANR-11-INBS-0010, www.metabohub.fr), for their help in collecting, processing, and interpreting 13C NMR and MS data. We also thank Marcus Persicke, Maud Heuillet, Edern Cahoreau, Lindsay Peyriga, Pierre Millard, Gilles Vieira, and Jean-Charles Portais for the insights they provided. This work was supported by ERA-CoBioTech’s project C1Pro (ANR-17-COBI-0003-05 and FNR-22023617).
Publisher Copyright:
Copyright © 2020 Delépine et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.
PY - 2020/10
Y1 - 2020/10
N2 - 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.
AB - 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.
KW - Bacillus methanolicus MGA3
KW - C metabolic flux analysis
KW - Methanol
KW - Natural methylotrophy
KW - Nonstationary MFA
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UR - https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=pure_univeritat_ramon_llull&SrcAuth=WosAPI&KeyUT=WOS:000579368300017&DestLinkType=FullRecord&DestApp=WOS_CPL
U2 - 10.1128/mSystems.00745-20
DO - 10.1128/mSystems.00745-20
M3 - Article
C2 - 32963101
AN - SCOPUS:85092273018
SN - 2379-5077
VL - 5
JO - mSystems
JF - mSystems
IS - 5
M1 - e0074520
ER -