Molecular modeling and redesign of alginate lyase from Pseudomonas aeruginosa for accelerating CRPA biofilm degradation.

Abstract

Administration of an efficient alginate lyase (AlgL) or AlgL mutant may be a promising therapeutic strategy for treatment of cystic fibrosis patients with Pseudomonas aeruginosa infections. Nevertheless, the catalytic activity of wild-type AlgL is not sufficiently high. It is highly desired to design and discover an AlgL mutant with significantly improved catalytic efficiency against alginate substrates. For the purpose of identifying an AlgL mutant with significantly improved catalytic activity, in this study, we first constructed and validated a structural model of AlgL interacting with substrate, providing a better understanding of the interactions between AlgL and its substrate. Based on the modeling insights, further enzyme redesign and experimental testing led to discovery of AlgL mutants, including the K197D/K321A mutant, with significantly improved catalytic activities against alginate and acetylated alginate in ciprofloxacin-resistant P. aeruginosa (CRPA) biofilms. Further anti-biofilm activity assays have confirmed that the K197D/K321A mutant with piperacillin/tazobactam is indeed effective in degrading the CRPA biofilms. Co-administration of the potent mutant AlgL and an antibiotic (such as a nebulizer) could be effective for therapeutic treatment of CRPA-infected patients with cystic fibrosis. Proteins 2016; 84:1875-1887. © 2016 Wiley Periodicals, Inc.

• Authors: Hoon Cho, Xiaoqin Huang, Yu Lan Piao, Da Eun Kim, So Yeon Lee, Eun Jeong Yoon, So Hee Park, Kyoung Lee, Chul Ho Jang, Chang-Guo Zhan
• Journal: Proteins (Proteins) Vol. 84 Issue 12 Pg. 1875-1887 (12 2016) ISSN: 1097-0134 [Electronic] United States
• PMID: 27676452 (Publication Type: Journal Article)
• Copyright: © 2016 Wiley Periodicals, Inc.

Chemical References

• Alginates
• Anti-Bacterial Agents
• Bacterial Proteins
• Hexuronic Acids
• Recombinant Proteins
• Piperacillin, Tazobactam Drug Combination
• Ciprofloxacin
• Penicillanic Acid
• Glucuronic Acid
• Polysaccharide-Lyases
• poly(beta-D-mannuronate) lyase
• Piperacillin

Topics

• Acetylation
• Alginates (chemistry, metabolism)
• Amino Acid Sequence
• Anti-Bacterial Agents (pharmacology)
• Bacterial Proteins (genetics, metabolism, pharmacology)
• Biocatalysis
• Biofilms (drug effects, growth & development)
• Ciprofloxacin (pharmacology)
• Cloning, Molecular
• Drug Resistance, Bacterial (drug effects)
• Drug Synergism
• Drug Therapy, Combination
• Escherichia coli (genetics, metabolism)
• Gene Expression
• Glucuronic Acid (chemistry, metabolism)
• Hexuronic Acids (chemistry, metabolism)
• Hydrolysis
• Kinetics
• Molecular Dynamics Simulation
• Mutation
• Penicillanic Acid (analogs & derivatives, pharmacology)
• Piperacillin (pharmacology)
• Piperacillin, Tazobactam Drug Combination
• Polysaccharide-Lyases (genetics, metabolism, pharmacology)
• Protein Domains
• Protein Engineering
• Protein Structure, Secondary
• Pseudomonas aeruginosa (chemistry, drug effects, enzymology, growth & development)
• Recombinant Proteins (genetics, metabolism, pharmacology)
• Sequence Alignment
• Structural Homology, Protein