Alkaline protease mutant, and gene, engineered strain, preparation method and application thereof

US11655464B2 · kind B2 · utility

Assignees

• Tianjin University of Science and Technology · CN
• Shandong Lonct Enzymes Co. LTD · CN

Inventors

• Fuping Lu · Tianjin, CN
• Yihan Liu · Tianjin, CN
• Yu Li · Shanghai, CN
• Xingji WANG · Linyi, CN
• Kefen Wang · Linyi, CN
• Wenlong Liu · Qingdao, CN
• Fufeng Liu · Tianjin, CN
• Huitu Zhang · Tianjin, CN

Key dates

• Filing date: Jul 31, 2022
• Grant date: May 23, 2023
• Expiry date: Jul 31, 2042

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N9/54
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

An alkaline protease mutant, and a gene, engineered strain, a preparation method and application thereof are provided. The method comprises the following steps of extracting genome DNA of Bacillus clausii, performing PCR amplification to obtain a wild-type alkaline protease gene sequence, mutating the wild-type alkaline protease gene obtained by the amplification through an error-prone PCR, performing high-throughput screening to obtain a plurality of highly active alkaline protease genes, performing DNA shuffling on the highly active alkaline protease genes, and performing screening to obtain eight alkaline protease mutant genes with higher activity.