Schultenkämper, Kerstin; Gütle, Desirée D.; Gil Lopez, Marina; Keller, Laura B.; Zhang, Lin; Einsle, Oliver; Jacquot, Jean-Pierre; Wendisch, Volker F.: Interrogating the role of the two distinct fructose-bisphosphate aldolases of Bacillus methanolicus by site-directed mutagenesis of key amino acids and [...]. In: Front Microbiol. Jg.12. 2021
Inhalt
- Introduction
- Materials and Methods
- Structure Determination and Analysis
- Strains, Media, and Culture Conditions
- Recombinant DNA Work
- CRISPRi Targeting of fba C and of fba P
- Protein Overproduction and Purification
- Enzyme Activity Assays
- Gene Expression Analysis Using qRT-PCR
- Results
- Structural Comparison of Native FBA C and FBA P
- Influence of Amino Acid Residues 51 and 140 of FBA P and FBA C on Their Glycolytic Aldol Cleavage Activities
- Influence of Amino Acid Residues 51 and 140 of FBA P and FBA C on Their Gluconeogenic Aldol Condensation Activities
- Repression of fba C and fba P by CRISPR Interference
- Discussion
- Data Availability Statement
- Author Contributions
- Supplementary Material
- References