Markert, Benno; Stolzenberger, Jessica; Brautaset, Trygve; Wendisch, Volker F.: Characterization of two transketolases encoded on the chromosome and the plasmid pBM19 of the facultative ribulose monophosphate cycle methylotroph Bacillus [...]. In: BMC Microbiology. Jg.14 H. 1. 2014
Inhalt
- Background
- Results
- Bioinformatic analysis and phylogeny of the TKTP and TKTC from B. methanolicus
- Overexpression of tktC and tktP resulted in increased TKT activity in B. methanolicus
- Heterologous expression, purification and biochemical characterization of the TKTP and TKTC
- (I) Overexpression, purification and molecular mass detection
- (II) Determining the optimal conditions for TKT activity
- (III) TKTC displays higher temperature stability than TKTP
- (IV) Formation of the TKT apoform and reconstitution of the holoenzyme revealed a bivalent metal ion dependency for activity
- (V) TKT activities are inhibited by ATP, ADP, EDTA and Ni2+
- TKTP and TKTC share similar kinetic parameters and substrate spectrum
- Discussion
- Conclusion
- Methods
- Microorganisms and cultivation conditions
- Homologous overexpression of tktC and tktP in B. methanolicus
- Purification molecular mass determination of TKT proteins
- Enzyme assays for the purified TKT proteins
- Formation of the transketolase apoform and reconstitution of the holoenzyme
- Computational analysis
- Abbreviations
- Authors' contribution
- Author details
- References