Thermostability is a key factor for the industrial application of enzymes. This review categorizes enzymes by their applications and discusses the importance of engineering thermostability for practical use. It summarizes fundamental theories and recent advancements in enzyme thermostability modification, including directed evolution, semi-rational design, and rational design. Directed evolution uses high-throughput screening to generate random mutations, while semi-rational design combines hotspot identification with screening. Rational design focuses on key residues to enhance stability by improving rigidity, foldability, and reducing aggregation. The review also covers rational strategies like engineering folding energy, surface charge, machine learning methods, and consensus design, along with tools that support these approaches. Practical examples are critically assessed to highlight the benefits and limitations of these strategies. Finally, the challenges and potential contributions of artificial intelligence in enzyme thermostability engineering are discussed.
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http://dx.doi.org/10.1016/j.ijbiomac.2024.139067 | DOI Listing |
Int J Biol Macromol
December 2024
Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology and School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China; Science Center for Future Foods, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China. Electronic address:
Thermostability is a key factor for the industrial application of enzymes. This review categorizes enzymes by their applications and discusses the importance of engineering thermostability for practical use. It summarizes fundamental theories and recent advancements in enzyme thermostability modification, including directed evolution, semi-rational design, and rational design.
View Article and Find Full Text PDFJ Agric Food Chem
December 2024
College of Light Industry and Food Engineering, Guangxi University, 100 Daxue Road, Nanning 530004, Guangxi, China.
d-Allulose 3-epimerase (DAEase) derived from has excellent properties in the catalytic production of d-allulose, a rare sugar with unique biological functions. However, the industrial application of DAEase (Cb-DAEase) for d-allulose production is hindered by its low enzyme activity, poor long-term thermostability, and pH tolerance. In this study, we identified potential noncatalytic residues in Cb-DAEase using methods such as proline substitution, surface charge engineering, and surface residue prediction.
View Article and Find Full Text PDFBiophys Chem
December 2024
School of Food Science and Engineering, Ocean University of China, Qingdao, Shandong 266500, China.
Glucose isomerase is generally used in the industrial production of high-fructose corn syrup, and a heat- and acid-resistant glucose isomerase is preferred. However, most glucose isomerases exhibit low activity or inactivation at low pH. In this study, we demonstrated that two combination mutants formed by introducing positive and negative charges near the active site and on the surface of the enzyme demonstrated a successful reduction in the optimal pH and increase in the specific activity of glucose isomerase from Thermotoga maritima (TMGI).
View Article and Find Full Text PDFJ Appl Glycosci (1999)
November 2024
1 Matsutani Chemical Industry Co., Ltd.
D-Allulose 3-epimerase catalyzes C-3 epimerization between D-fructose and D-allulose was found in strain M30. The enzyme gene was cloned, and its recombinant enzyme and the mutant variants were expressed in Using the information of the sequence and model structure, we succeed in the improvement of melting temperature for the enzyme without significant loss of the enzyme activity by protein engineering method. The melting temperatures were increased by 2.
View Article and Find Full Text PDFActa Biochim Biophys Sin (Shanghai)
December 2024
Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
The use of green fluorescence protein (GFP) has advanced numerous areas of life sciences. An ultra-thermostable GFP (TGP), engineered from a coral GFP, offers potential advantages over traditional jellyfish-derived GFP because of its high stability. However, owing to its later discovery, TGP lacks the extensive toolsets available for GFP, such as heavy chain-only antibody binders known as nanobodies.
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