Artificially designed penicillin G acylase proenzyme and coding sequence and applications thereof

• Main Authors: CAO CHUNLAI, LIANG XIONGJI, LIU HEDONG, HUANG GUOZHOU
• Format: Patent
• Language: Chinese; English
• Published: 16.10.2018
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses artificially designed penicillin G acylase proenzyme and a coding sequence and applications thereof. Substrate combining steric effects can be reduced and the substrate can beurged to rapidly enter an active pocket by mutating more than one of large side chain amino acids near a substrate binding pocket of wild penicillin G acylase proenzyme into side chain amino acids which are smaller than the amino acids before mutation; and at the same time, through the mutation of more than one of key hydrophilic amino acids or weakly hydrophobic amino acids in the active pocket near a substrate-active center into the amino acids whose hydrophobicity is stronger than that of amino acids before the mutation, the hydrophobicity around the substrate binding pocket can be increased, the combination of water molecules can be reduced, hydrolysis can be reduced, S/H can be increased from reducing hydrolysis and increasing synthesis, and the capability of synthesizing penicillin Gacylase into amoxicillin