Echinocandin B deacylase (EBDA), from Actinoplanes utahensis ZJB-08196, is capable of cleaving the linoleoyl group from echinocandin B (ECB), forming the echinocandin B nucleus (ECBN), which is a key precursor of semisynthetic antifungal antibiotics. In the present study, molecular evolution of AuEBDA by random mutagenesis combined with site-directed mutagenesis (SDM) and screening was performed. Random mutagenesis on the wild-type (WT) AuEBDA generated two beneficial substitutions of G287Q, R527V. The "best" variant AuEBDA-G287Q/R527V was obtained by combining G287Q with R527V through SDM, which was most active at 35 °C, pH 7.5, with K and v values of 0.68 mM and 395.26 U/mg, respectively. Mutation of G287Q/R527V markedly increased the catalytic efficiency k/K by 290% compared with the WT-AuEBDA.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/s12010-019-03170-3 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Genome-Wide A → G and C → T Mutations Induced by Functional TadA Variants in .
ACS Synth Biol
January 2025
Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, P. R. China.
The fusion expression of deoxyribonucleic acid (DNA) replication-related proteins with nucleotide deaminase enzymes promotes random mutations in bacterial genomes, thereby increasing genetic diversity among the population. Most previous studies have focused on cytosine deaminase, which produces only C → T mutations, significantly limiting the variety of mutation types. In this study, we developed a fusion expression system by combining DnaG (RNA primase) with adenine deaminase TadA-8e (DnaG-TadA) in , which is capable of rapidly introducing A → G mutations into the genome, resulting in a 664-fold increase in terms of mutation rate.
View Article and Find Full Text PDFMetabolic engineering of Priestia megaterium for 2'-fucosyllactose production.
Microb Cell Fact
January 2025
Department of Chemical & Biological Engineering, Korea University, Seoul, 136-763, Korea.
Background: 2'-Fucosyllactose (2'-FL) is a predominant human milk oligosaccharide that significantly enhances infant nutrition and immune health. This study addresses the need for a safe and economical production of 2'-FL by employing Generally Recognized As Safe (GRAS) microbial strain, Priestia megaterium ATCC 14581. This strain was chosen for its robust growth and established safety profile and attributing suitable for industrial-scale production.
View Article and Find Full Text PDFEnhanced cyanophycin accumulation in diazotrophic cyanobacterium through random mutagenesis and tailored selection under varying phosphorus availability.
Bioresour Technol
December 2024
Microalgal Biotechnology Laboratory, The French Associates Institute for Agriculture and Biotechnology of Drylands, The J. Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, Sede Boqer Campus, 8499000, Israel. Electronic address:
This study explored a sustainable alternative to the Haber-Bosch process by enhancing production of nitrogen-rich polymer cyanophycin (CGP) in the diazotrophic cyanobacterium Nostoc sp. PCC7120. Applying UV-mutagenesis followed by canavanine selection, we isolate an initial mutant with enhanced CGP accumulation.
View Article and Find Full Text PDFOptimizing enzyme thermostability by combining multiple mutations using protein language model.
mLife
December 2024
State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai China.
Article Synopsis
- Optimizing enzyme thermostability is crucial for protein science and industry, but combining multiple mutations can lead to inactivation, making traditional methods slow and inefficient.
- Researchers developed an AI-driven method to enhance enzyme thermostability by efficiently recombining beneficial single-point mutations, using data from various mutant groups.
- After two design rounds, the study achieved 50 combinatorial mutants with 100% success, including one exceptional mutant that significantly increased melting temperature and half-life, while also revealing complex interactions (epistasis) among mutations.
Directed evolution of glutamate decarboxylase B for enhancing its enzyme activity towards nearly neutral pHs based on error-prone PCR.
Int J Biol Macromol
December 2024
College of Food Science and Biotechnology, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou, Zhejiang Province 310035, People's Republic of China. Electronic address:
Glutamate decarboxylases (GADs) can catalyze the conversion of l-glutamate to γ-aminobutyric acid (GABA), while consuming one H. However, the GADs found so far are catalytically active in the pHs of 3.8-5.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!