Article
Biochemistry & Molecular Biology
Qiuya Gu, Yu Lu, Jianli Zhou, Wenhua Yang, Ke Wang, Xiaobo Liu, Xiaobin Yu
Summary: Combinational mutagenesis was used to improve the catalytic efficiency and adaptability of alginate lyase from Pseudoalteromonas sp. Alg6B, which showed significant potential for the production of AOS from marine alginate.
PROCESS BIOCHEMISTRY
(2023)
Article
Biotechnology & Applied Microbiology
Li-Qun Jin, Yi-Ting Jin, Jing-Wei Zhang, Zhi-Qiang Liu, Yu-Guo Zheng
Summary: Glucose isomerase mutant TEGI-M2 showed optimal activity at high temperature and neutral pH, with structural differences compared to the original enzyme investigated through molecular docking. TEGI-M2 exhibited increased enzyme activity and decreased Km, resulting in higher D-fructose yield in one-step biosynthesis of HFCS. This improved catalytic performance of TEGI-M2 is significant for industrial production of D-fructose.
ENZYME AND MICROBIAL TECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Sachin Kumar, Vijay Kumar Bhardwaj, Shweta Guleria, Rituraj Purohit, Sanjay Kumar
Summary: This study utilized a structure-guided consensus approach to improve the enzymatic properties of Cu,Zn SOD from the high-altitude plant Potentilla atrosanguinea. The T97D substitution in SOD led to improved dimer stability and catalytic efficiency compared to the wild-type (WT), as validated by computational and experimental tools. This mutation not only enhanced enzyme thermostability but also laid the groundwork for designing SODs with improved kinetics by exploring interface residues as potential targets.
BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS
(2022)
Article
Biochemistry & Molecular Biology
Satoru Watanabe, Masahiro Ito, Takanori Kigawa
Summary: DiRect is a novel PCR-based mutagenesis method that achieves high quality product (>99% substitution) without the use of recombinant DNA technology. It has been successfully applied in the study of an industrially relevant enzyme, showing simplicity, efficiency, and potential scalability in site-directed mutagenesis.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
(2021)
Article
Food Science & Technology
Huabiao Miao, Xia Xiang, Nanyu Han, Qian Wu, Zunxi Huang
Summary: This study aimed to improve the thermostability of the serine protease PB92 from Bacillus alcalophilus to meet the high-temperature requirements of biotechnological treatments. Mutations were introduced at eight sites, and 21 mutants were constructed, among which 15 mutants showed increased half-life values at 65 degrees C. Six complex mutants with improved thermal stability were identified based on enzyme activity and thermostability scores, and structural analysis suggested that the increased stability might be due to additional hydrophobic interactions and reduced flexibility. Overall, the N18L/R143L/S97A, N18L/R143L/S99L, and N18L/R143L/G100A mutants showed 4-fold increase in thermal stability, indicating potential industrial applications.
Article
Agriculture, Multidisciplinary
Cheng-Hua Wang, Wu-Ping Xiong, Cheng Huang, Xiao-Ming Li, Qing-Yan Wang, Ri-Bo Huang
Summary: This study successfully increased the catalytic activity and efficiency of an exoinulinase under acidic conditions through rational site-directed mutagenesis. The mutants showed improved substrate preference for inulin and may serve as promising candidates for industrial inulin hydrolysis. The study demonstrates the potential of mutating non-conserved substrate recognition residues for industrial enzyme improvements.
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
(2021)
Article
Biochemistry & Molecular Biology
Sitian Gu, Xiaojun Dai, Jiang Jiang, Yuanfa Liu
Summary: In this study, three single-site mutations were introduced into OaCLH to improve its catalytic efficiency. The mutants W160R and V228C showed significantly increased enzyme affinity and catalytic activity against all substrates, while the D224 N mutant changed its preferred substrate. These findings provide structural basis information for the catalytic activity and substrate specificity of OaCLH.
PROCESS BIOCHEMISTRY
(2022)
Article
Biotechnology & Applied Microbiology
Hai-Yan Zhou, Xiao-Nan Yi, Qi Chen, Jian-Bao Zhou, Shu-Fang Li, Xue Cai, De-Shui Chen, Xin-Ping Cheng, Mian Li, Hong-Yan Wang, Kai-Qian Chen, Zhi-Qiang Liu, Yu-Guo Zheng
Summary: In this study, a novel endoglucanase (CgEndo) from Colletotrichum graminicola was modified by site-directed mutagenesis to improve its catalytic efficiency. Two mutants, Y63S and N20D/S113T, showed significantly increased enzyme activities. The mutations improved the proximity between the enzyme and its substrate, leading to higher enzyme activities and catalysis efficiency. These findings lay an important foundation for further engineering of the endoglucanase and its practical application in efficient degradation of cellulosic biomass.
ENZYME AND MICROBIAL TECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Kemin Lv, Wenyu Shao, Marcelo Monteiro Pedroso, Jiayu Peng, Bin Wu, Jiahuang Li, Bingfang He, Gerhard Schenk
Summary: In this study, the processive endoglucanase EG5C-1 from Bacillus subtilis was used as the basis for enzyme engineering, resulting in the discovery of variants D70Q and S235W, as well as the double mutant D70Q/S235W, which showed significantly improved activity towards substrates CMC and Avicel. Kinetic measurements demonstrated that the double mutant had higher substrate affinity and catalytic efficiency.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Biotechnology & Applied Microbiology
Zhanqing Fan, Li Fang, Limei Wu, Zheren Wang, Yanan Wang, Caijing Han, Xiaoting Liu
Summary: This study aimed to improve the catalytic activity of aspartate kinase (AK), the first key enzyme in the aspartic acid metabolism pathway, and weaken the feedback inhibition of metabolites. The mutant A380M with significantly increased enzyme activity showed enhanced substrate affinity. Molecular dynamics simulations revealed changes in the binding force and interaction between AK and its substrates.
BIOPROCESS AND BIOSYSTEMS ENGINEERING
(2022)
Article
Green & Sustainable Science & Technology
Fatima Akram, Ikram ul Haq, Amna Aqeel, Zeeshan Ahmed, Fatima Iftikhar Shah
Summary: Human culture has been exploring the potential of using hyperthermophiles and their enzymes as a bio-competitive alternative to fossil-based fuel resources; research has characterized cellulolytic glycoside hydrolases and discussed engineering techniques to enhance the production and stability of thermostable cellulolytic enzymes.
RENEWABLE & SUSTAINABLE ENERGY REVIEWS
(2021)
Article
Biotechnology & Applied Microbiology
Guanglu Wang, Mengyuan Wang, Lanxi Liu, Xiaohan Hui, Bingyang Wang, Ke Ma, Xuepeng Yang
Summary: By site-directed mutagenesis of glycerol kinase gene in Bacillus subtilis, the glycerol utilization efficiency was improved, leading to increased growth rate and biomass.
BIOTECHNOLOGY LETTERS
(2022)
Article
Biotechnology & Applied Microbiology
Qi Li, Xinyi Tong, Yunpeng Jiang, Dongdong Li, Linguo Zhao
Summary: This study successfully enhanced the xylose tolerance of GH39 family beta-xylosidase Xln-DT through gene mutation, significantly improving its activity and thermal stability.
ENZYME AND MICROBIAL TECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Zhongxiu Chen, Longbin Wang, Yuyu Shen, Dunji Hu, Liying Zhou, Fuping Lu, Ming Li
Summary: Recombinant genes GA1 and GA2 were expressed in Saccharomyces cerevisiae, showing higher thermostability and catalytic efficiency compared to the original genes.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Hongguan Xing, Pingping Wang, Xing Yan, Yi Yang, Xinliang Li, Rui Liu, Zhihua Zhou
Summary: Directed evolution was used to enhance the thermostability of phytase, resulting in the generation of five mutants with higher residual activities and catalytic efficiencies. The substitution of C75 residue was identified as the main reason for the improved thermostability, indicating the crucial role of this amino acid in the stability and catalytic efficiency of phytase.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)