Novel beta-xylosidase and preparation thereof

• Main Authors: GE XIUQI, WANG FENGHUA, LIU YIHAN, LU FUPING
• Format: Patent
• Language: Chinese; English
• Published: 22.12.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention belongs to the technical field of genetic engineering of enzymes, and particularly relates to a beta-xylosidase mutant with improved enzyme activity and preparation thereof. According tothe invention, a wild type beta-xylosidase gene of Bacillus pumilus is obtained through a molecular biological technology means, random mutation is carried out on the wild type beta-xylosidase gene by utilizing an error-prone PCR technology to obtain a beta-xylosidase mutant E455G and a coding gene xylm thereof, recombinant plasmids are reconstructed, efficient expression of the recombinant plasmids in Bacillus subtilis, Bacillus licheniformis and Bacillus amyloliquefaciens is realized, and high-activity beta-xylosidase is obtained through technologies such as fermentation and extraction. 本发明属于酶的基因工程技术领域，具体涉及一种酶活力提高的β-木糖苷酶突变体及其制备。本发明通过分子生物学技术手段获得短小芽孢杆菌(Bacillus pumilus)野生型的β-木糖苷酶基因，利用易错PCR技术对野生型β-木糖苷酶基因进行随机突变，得到β-木糖苷酶突变体E455G，及其编码基因xylm，重新构建重组质粒，并实现了其在枯草芽孢杆菌、地衣芽孢杆菌和解淀粉芽孢杆菌中的高效表达，通过发酵、提取等技术获得高活力的β-木糖苷酶。