Targeting Efficient Features of Urate Oxidase to Increase Its Solubility.

Autor: Rahbar MR; Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran., Nezafat N; Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.; Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, P.O. Box 71345-1583, Shiraz, Iran., Morowvat MH; Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.; Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, P.O. Box 71345-1583, Shiraz, Iran., Savardashtaki A; Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran., Ghoshoon MB; Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.; Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, P.O. Box 71345-1583, Shiraz, Iran., Mehrabani-Zeinabad K; Department of Biostatistics, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran., Ghasemi Y; Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. ghasemiy@sums.ac.ir.; Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, P.O. Box 71345-1583, Shiraz, Iran. ghasemiy@sums.ac.ir.
Jazyk: angličtina
Zdroj: Applied biochemistry and biotechnology [Appl Biochem Biotechnol] 2024 Feb 03. Date of Electronic Publication: 2024 Feb 03.
DOI: 10.1007/s12010-023-04819-w
Abstrakt: With the demand for mass production of protein drugs, solubility has become a serious issue. Extrinsic and intrinsic factors both affect this property. A homotetrameric cofactor-free urate oxidase (UOX) is not sufficiently soluble. To engineer UOX for optimum solubility, it is important to identify the most effective factor that influences solubility. The most effective feature to target for protein engineering was determined by measuring various solubility-related factors of UOX. A large library of homologous sequences was obtained from the databases. The data was reduced to six enzymes from different organisms. On the basis of various sequence- and structure-derived elements, the most and the least soluble enzymes were defined. To determine the best protein engineering target for modification, features of the most and least soluble enzymes were compared. Metabacillus fastidiosus UOX was the most soluble enzyme, while Agrobacterium globiformis UOX was the least soluble. According to the comparison-constant method, positive surface patches caused by arginine residue distribution are appropriate targets for modification. Two Arg to Ala mutations were introduced to the least soluble enzyme to test this hypothesis. These mutations significantly enhanced the mutant's solubility. While different algorithms produced conflicting results, it was difficult to determine which proteins were most and least soluble. Solubility prediction requires multiple algorithms based on these controversies. Protein surfaces should be investigated regionally rather than globally, and both sequence and structural data should be considered. Several other biotechnological products could be engineered using the data reduction and comparison-constant methods used in this study.
(© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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