Pfeifenschneider, Johannes; Markert, Benno; Stolzenberger, Jessica; Brautaset, Trygve; Wendisch, Volker F.: Transaldolase in Bacillus methanolicus: Biochemical Characterization and Biological Role in Ribulose Monophosphate Cycle. In: BMC Microbiology. Jg.20. 2020
Inhalt
- Results
- The genomes from B. methanolicus PB1 and MGA3 encode transaldolase
- Enzyme assays in crude extracts demonstrated Ta activities for both strains
- Construction of mutant taMGA3 genes expressing full-length Ta proteins
- Heterologous expression of different MGA3 ta variants in E. coli
- Overexpression and purification of the TaMGA3rec and TaPB1 proteins
- Biochemical characterization of TaPB1
- Heterologous expression of taPB1 and TaMGA3rec complemented Ta-deficiency in C. glutamicum ∆tal
- Overexpression of taPB1 in B. methanolicus MGA3
- Size exclusion chromatography, SDS-PAGE and MALDI-TOF analysis of MGA3 crude extracts
- Identification of a possible transcription start site upstream of the MGA3 ta gene
- Proteome data indicating biosynthesis of functional Ta encoded by taMGA3put
- Discussion
- Conclusions
- Methods
- Microorganisms and cultivation conditions
- Construction of tal (cg1776) deletion mutant in C. glutamicum
- Vector constructions for heterologous expression of taPB1 and taMGA3rec of B. methanolicus and homologous expression of talCG in C. glutamicum
- Vector construction for heterologous expression of different taMGA3 variants in E. coli
- Vector construction for homologous overexpression of taPB1 in B. methanolicus MGA3
- Purification and molecular mass determination of Ta proteins
- Enzyme assays for the purified Ta proteins
- Tryptic digestion of proteins and mass spectrometry analysis
- Computational analysis
- Abbreviations
- Acknowledgements
- Authors’ contributions
- Funding
- Availability of data and materials
- Ethics approval and consent to participate
- Consent for publication
- Competing interests
- Author details
- References
- Publisher’s Note