Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle

Pfeifenschneider J, Markert B, Stolzenberger J, Brautaset T, Wendisch VF (2020)
BMC Microbiology 20: 63.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
Background The Gram-positive facultative methylotrophic bacterium Bacillus methanolicus uses the sedoheptulose-1,7-bisphosphatase (SBPase) variant of the ribulose monophosphate (RuMP) cycle for growth on the C1 carbon source methanol. Previous genome sequencing of the physiologically different B. methanolicus wild-type strains MGA3 and PB1 has unraveled all putative RuMP cycle genes and later, several of the RuMP cycle enzymes of MGA3 have been biochemically characterized. In this study, the focus was on the characterization of the transaldolase (Ta) and its possible role in the RuMP cycle in B. methanolicus. Results The Ta genes of B. methanolicus MGA3 and PB1 were recombinantly expressed in Escherichia coli, and the gene products were purified and characterized. The PB1 Ta protein was found to be active as a homodimer with a molecular weight of 54 kDa and displayed KM of 0.74 mM and Vmax of 16.3 U/mg using Fructose-6 phosphate as the substrate. In contrast, the MGA3 Ta gene, which encodes a truncated Ta protein lacking 80 amino acids at the N-terminus, showed no Ta activity. Seven different mutant genes expressing various full-length MGA3 Ta proteins were constructed and all gene products displayed Ta activities. Moreover, MGA3 cells displayed Ta activities similar as PB1 cells in crude extracts. Conclusions While it is well established that B. methanolicus can use the SBPase variant of the RuMP cycle this study indicates that B. methanolicus possesses Ta activity and may also operate the Ta variant of the RuMP.
Stichworte
Methylotrophy; Transaldolase; RuMP cycle; Bacillus methanolicus
Erscheinungsjahr
2020
Zeitschriftentitel
BMC Microbiology
Band
20
Art.-Nr.
63
eISSN
1471-2180
Finanzierungs-Informationen
Open-Access-Publikationskosten wurden durch die Deutsche Forschungsgemeinschaft und die Universität Bielefeld gefördert.
Page URI
https://pub.uni-bielefeld.de/record/2941820

Zitieren

Pfeifenschneider J, Markert B, Stolzenberger J, Brautaset T, Wendisch VF. Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle. BMC Microbiology. 2020;20: 63.
Pfeifenschneider, J., Markert, B., Stolzenberger, J., Brautaset, T., & Wendisch, V. F. (2020). Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle. BMC Microbiology, 20, 63. https://doi.org/10.1186/s12866-020-01750-6
Pfeifenschneider, J., Markert, B., Stolzenberger, J., Brautaset, T., and Wendisch, V. F. (2020). Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle. BMC Microbiology 20:63.
Pfeifenschneider, J., et al., 2020. Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle. BMC Microbiology, 20: 63.
J. Pfeifenschneider, et al., “Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle”, BMC Microbiology, vol. 20, 2020, : 63.
Pfeifenschneider, J., Markert, B., Stolzenberger, J., Brautaset, T., Wendisch, V.F.: Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle. BMC Microbiology. 20, : 63 (2020).
Pfeifenschneider, Johannes, Markert, Benno, Stolzenberger, Jessica, Brautaset, Trygve, and Wendisch, Volker F. “Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle”. BMC Microbiology 20 (2020): 63.
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2020-04-09T09:17:25Z
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