Article
Biochemistry & Molecular Biology
Mohammed Hamed Alqarni, Ahmed Ibrahim Foudah, Magdy Mohamed Muharram, Haritium Budurian, Nikolaos E. Labrou
Summary: AMPS-HDB reacts covalently with CbFDH, leading to complete inactivation of the enzyme activity, while NAD(+) and NADH protect CbFDH from inactivation by AMPS-HDB. Molecular modelling studies identified Arg174 as the target for AMPS-HDB, confirmed by site-directed mutagenesis.
Article
Biochemical Research Methods
Wen-Ching Lin, Hao-Cheng Tang, Han Ying Wang, Chia-Yi Kao, You-Chiun Chang, Athena Hsu Li, Shi-Bing Yang, Kurt Yun Mou
Summary: This study introduces a unique mutagenesis method called the reverse Kunkel method, which combines the advantages of user-defined mutagenesis and random mutagenesis. By coupling with phage display and yeast display selections, the method successfully generates improved antibodies with enhanced affinity and immunostaining performance.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Jian Li, Qi-Long Hu, Zhendong Song, Albert S. C. Chan, Xiao-Feng Xiong
Summary: Site-selective modification of peptides/proteins is a crucial method for studying post-translational modifications, chemical biology, and drug development. In this study, a visible-light-driven and Cys-directed Lys site-selective stapling approach was developed, enabling selective modification and multi-functionalization.
SCIENCE CHINA-CHEMISTRY
(2022)
Article
Microbiology
Emily C. Hoedt, Francesca Bottacini, Nora Cash, Roger S. Bongers, Kees van Limpt, Kaouther Ben Amor, Jan Knol, John MacSharry, Douwe van Sinderen
Summary: Members of the genus Bifidobacterium are difficult to genetically manipulate due to their extensive and variable Restriction-Modification systems. A modified suicide vector, pFREM28, was developed to target specific genes in B. breve by removing known restriction sites, reducing the time, effort, and resources required for generating site-directed mutants. This approach could be applied to other (bifido)bacterial species as well.
FRONTIERS IN MICROBIOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Siying Xiang, Minsong Gao, Jie Cao, Xiao Shu, Mohan Cheng, Fengqin Wang, Ting Deng, Jianzhao Liu
Summary: The method utilizes allyl-substituted methyltransferase cofactor to label a specific base in an RNA substrate, enabling precise identification of the labeling site using mutation signals from standard nucleic acid sequencing. It provides a platform for studying enzymatic methylations in long and complex RNA substrates, as well as facilitating the discovery of new methyltransferases.
CHEMICAL COMMUNICATIONS
(2021)
Article
Agriculture, Multidisciplinary
Su-Jin Jun, Jung-A Lee, Young-Wan Kim, Sang-Ho Yoo
Summary: Through site-directed mutagenesis, a robust mutant Bifidobacterium thermophilum (BtAS) strain BtAS(Y414F/P200R) with enhanced turanose productivity was successfully developed, showing an increase in turanose yield by two times compared to the wild type BtAS and completing the reaction in half the time.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Soroosh Pouyan, Milad Lagzian, Mohammad Hossein Sangtarash
Summary: This study characterized a new alpha-amylase from Bacillus cereus GL96 and used an in-silico approach to redesign the enzyme for improved thermal stability. The engineered enzyme showed higher stability and activity at elevated temperatures compared to the wild-type enzyme. Specifically, the D162K mutation resulted in the best performance, with increased optimum temperature and improved stability. This mutant enzyme holds promise for further investigation and industrial applications.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2022)
Article
Plant Sciences
Xia Li, Geng Chen, Qing-Qing Gao, Chun-Fan Xiang, Cheng-Xiao Yuan, Xiao-Ning Li, Yan-Yu Shu, Guang-Hui Zhang, Yan-Li Liang, Sheng-Chao Yang, Chen-Xi Zhai, Yan Zhao
Summary: This study explored the key enzymes involved in cucurbitacins biosynthesis in the Chinese traditional medicinal plant Hemsleya chinensis. Using AlphaFold2 and molecular docking, the researchers successfully predicted the 3D structure of HcOSC6 and its binding features to 2,3-oxidosqualene. Mutagenesis experiments identified three key amino acid residues, E246, M261, and D490, which significantly affected the activity of HcOSC6. This study not only characterized three potential key residues for producing cucurbitacins, but also provided insights into their significant role in metabolic engineering.
FRONTIERS IN PLANT SCIENCE
(2023)
Article
Biochemistry & Molecular Biology
Lena Goshayeshi, Sara Yousefi Taemeh, Nima Dehdilani, Mohammadreza Nasiri, Mohammad M. Ghahramani Seno, Hesam Dehghani
Summary: The study demonstrates the use of an RNA-guided piggyBac transposase and dual sgRNAs for site-directed integrations in the human ROSA26 safe harbor region. This methodology shows promise for targeting specific genomic regions, such as in cancer cells for inserting sequences into target regions or activating tumor suppressor genes.
Article
Biochemistry & Molecular Biology
Deborah Giordano, Bernardina Scafuri, Luigi De Masi, Lucia Capasso, Viviana Maresca, Lucia Altucci, Angela Nebbioso, Angelo Facchiano, Paola Bontempo
Summary: This study aimed to investigate the effects of Cambinol and its underlying mechanisms on cell differentiation. The findings demonstrated that Cambinol induced differentiation in MCF-7, NB4, and 3T3-L1 cell lines by modulating the expression of cell cycle-related proteins and nuclear receptors. In addition, molecular simulations revealed that Cambinol showed inhibitory effects on human SIRT1 and SIRT2, with stronger interaction with the substrate binding site of SIRT1. Overall, Cambinol may serve as a potential alternative for epigenetic therapies targeting SIRTs.
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)
Review
Plant Sciences
Olawale Samuel Adeyinka, Bushra Tabassum, Brice Landry Koloko, Ifedayo Victor Ogungbe
Summary: The improvement of CRISPR-Cas gene editing technology has facilitated genetic modification of desirable agronomic traits in crops, such as resistance to pathogens, drought tolerance, nutritional value, and yield-related traits. This presents a significant opportunity for precise genetic diversification in plants.
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)
Review
Chemistry, Organic
Houchao Xu, Jeroen S. Dickschat
Summary: In the past three decades, numerous terpene synthases have been characterized from various life kingdoms. The mechanisms of terpene synthases have been investigated through site-directed mutagenesis, providing valuable insights into their catalysis and resulting in mutants with improved yields.
SYNTHESIS-STUTTGART
(2022)
Article
Biotechnology & Applied Microbiology
Sitipon Leerawatthanakun, Thanapon Charoenwongpaiboon, Methus Klaewkla, Surasak Chunsrivirot, Jitnapa Sirirak, Tanin-ek Sriwitool, Karan Wangpaiboon, Rath Pichyangkura
Summary: In this study, the beta-mannanase from Bacillus subtilis HM7 (Man26HM7) isolated from Dynastes hercules larvae excrement was cloned and expressed. It exhibited excellent surfactant stability and has potential applications in detergent and cleaning products. Mutagenesis and computational analysis were performed to enhance the surfactant stability of beta-mannanase.
ENZYME AND MICROBIAL TECHNOLOGY
(2022)