Alkaline protease mutant with improved stability and application thereof

• Main Authors: LI YU, LIU FUFENG, ZHANG HUITU, LIU YIHAN, WANG HONGBIN, LU FUPING, GUO ENPING
• Format: Patent
• Language: Chinese; English
• Published: 27.10.2023
• Subjects: ANIMAL AND VEGETABLE OILS, FATS, FATTY SUBSTANCES AND WAXES; BEER; BIOCHEMISTRY; CANDLES; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; DETERGENT COMPOSITIONS; DETERGENTS; ENZYMOLOGY; FATTY ACIDS THEREFROM; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; RECOVERY OF GLYCEROL; RESIN SOAPS; SOAP OR SOAP-MAKING; SPIRITS; USE OF SINGLE SUBSTANCES AS DETERGENTS; VINEGAR; WINE

The invention belongs to the technical field of bioengineering, and particularly relates to alkaline protease mutants which are obtained by random mutation through an error-prone PCR technology and have improved stability in two negative surfactants and application of the alkaline protease mutants. The alkaline protease mutant is a G95P/A96D/S99W/S101 T/P127S/S126T/N212S mutant, and the amino acid sequence of the alkaline protease mutant is as shown in SEQ ID NO. 5. The residual enzyme activity reaches 85.9% after 12 h in the presence of 2% alkyl benzene sulfonic acid AES, and the residual enzyme activity reaches 61.2% after 4 h in the presence of 2% alkyl benzene sulfonic acid LAS. The problem of low stability of the alkaline protease in the liquid washing liquid can be effectively solved. 本发明属于生物工程技术领域，具体涉及通过易错PCR技术进行随机突变获得在两种阴表面活性剂中稳定性提高的碱性蛋白酶突变体及其应用。所述碱性蛋白酶突变体为G95P/A96D/S99W/S101T/P127S/S126T/N212S突变体，氨基酸序列如SEQ ID NO.5所示。在2％烷基苯磺酸AES存在情况下12h后的残余酶活达到85.9％，在2％烷基苯磺酸LAS存在情况下4h后的残余酶活达到61.2％。能够有效解决碱性蛋白酶在液体洗涤液中稳定