Clarification of the Organic Solvent-Stability of Enzymes Based on Protein Engineering and Development of Organic Solvent-Stable Enzymes

点击翻译按钮获取中文摘要

Enzymes exhibit high substrate and reaction specificities as catalysts under mild conditions. Therefore, if enzymes are stable in the presence of organic solvents, they can be applied to develop environmentally friendly synthetic processes of fine-chemicals. In this research project, the clarification of the organic solvent-stability of enzymes based on protein engineering and molecular evolution engineering and fundamental research to develop organic solvent-stable enzymes were performed. The examined item in this research is as follows.1) Growth of organic solvent-tolerant Pseudomonas aeruginosa LST-03 in the presence of an organic solvent and production of organic solvent-stable enzymes2) Purification and characterization of organic solvent-stable LST-03 lipase3) Cloning and sequence of the gene of organic solvent-stable LST-03 lipase4) Cloning and sequence of the lipase genes of Pseudomonas aeruginosa LST-035) Purification and characterization of organic solvent-stable PST-01 protease6) Peptide synthesis catalyzed by PST-01 protease7) Conformational change of enzymes in the presence of an organic solvent8) Cloning and sequence of the gene of organic solvent-stable PST-01 protease9) Role of disulfide bonds of organic solvent-stable PST-01 protease10) Clarification of the organic solvent-stability of enzymes based on molecular evolution engineering