Structural characterization of an L-fuculose-1-phosphate aldolase from Klebsiella pneumoniae

• Published in: Biochemical and biophysical research communications 2022-06, Vol.607, p.15-19
• Main Authors: Lou, Xiaorui, Zhang, Jianyu, Liu, Shimeng, Wang, Runhao, Li, Weiping, Liu, Ruihua, Zhang, Qionglin, Bartlam, Mark
• Format: Article
• Language: English
• Subjects: Aldehyde-Lyases - metabolism; Catalysis; Crystal structure; Enzyme activity; Fructose-Bisphosphate Aldolase - chemistry; Fuculose 1-phosphate aldolase; Klebsiella pneumoniae; Klebsiella pneumoniae - metabolism
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2022.03.127

Fuculose phosphate aldolases play an important role in glycolysis and gluconeogenesis pathways. L-fuculose 1-phosphate aldolase catalyzes the reversible cleavage of L-fuculose 1-phosphate to DHAP and L-lactaldehyde. Class II aldolases found in bacteria are linked to pathogenesis of human pathogens, and have potential applications in the biosynthesis of carbohydrates and other chiral compounds. Here we report the structure of a putative L-fuculose 1-phosphate aldolase (KpFucA) from the nosocomial pathogen Klebsiella pneumoniae to 1.85 Å resolution. The enzyme crystallizes in space group P422 with one monomer per asymmetric unit. Analytical ultracentrifugation analysis confirms that KpFucA is a tetramer in solution. A magnesium ion cofactor and sulfate ion were identified in the active pocket. Enzyme activity assays confirmed that KpFcuA has a strong preference for L-fuculose 1-phosphate as a substrate, but can also catalyze the cleavage of fructose-1,6-bisphosphate and glucose-6-phosphate. This work should provide a starting point for further investigation of the role of KpFucA in K. pneumoniae pathogenesis or in industrial applications. •The crystal structure of the putative Klebsiella pneumoniae L-fuculose 1-phosphate aldolase (KpFucA) was determined.•KpFucA forms a stable tetramer in solution.•The structure reveals a class II aldolase, including a metal cofactor and a conserved His-His-His catalytic triad.•KpFcuA has a strong preference for L-fuculose 1-phosphate as a substrate.•This KpFcuA structure provides a starting point for further investigation of its role in K. pneumoniae pathogenesis.