{"title":"Purification, biochemical characterization and structural analysis of uricase produced from a new strain of Streptomyces.","authors":"Anand Kumar Dakuri, Girijasankar Guntuku, Swathi Nageswara, Sowjanya Gurugubelli, Jagadeesh Chiluvuru","doi":"10.1007/s10529-026-03717-5","DOIUrl":null,"url":null,"abstract":"<p><p>The isolate A85 was identified as Streptomyces enissocaesilis, and it might serve as a novel actinobacteria source for uricase production. The uricase was successfully produced and purified from this novel strain, demonstrating that it could be an efficient source for future applications. Characterization studies of the purified enzyme showed maximal activity at pH 9 and 37 °C. From the differential scanning calorimetry, a melting point of 55.6 °C was observed, which ensured that the purified uricase is thermostable. Overall, 32 fold purification with a 48% yield of uricase was recovered at the end of purification steps. Inhibitor studies revealed that the purified uricase is metalloprotease. V<sub>max</sub> and K<sub>m</sub> values were found to be 30.39 U/mL and 0.0212 mM, respectively. The molecular mass of the purified uricase was found to be around 32 KDa. Further MALDI-TOF MS analysis identified four peptides as uricase, with a significant score. The purified enzyme was classified as an uricase based on a comprehensive analysis that integrated its amino acid sequence and conserved regions. The secondary structure of the purified uricase using PDBsum revealed 1 beta sheet, 1 beta hairpin, 1 beta bulge, 2 strands, and 2 beta turns. The 3D structure of the purified uricase was predicted through homology modeling, and the structure was validated using Ramachandran plot analysis. Docking results revealed significant binding affinity of -4.3 kcal/mol, which indicates strong and stable interaction of uricase and uric acid substrate.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 3","pages":""},"PeriodicalIF":2.5000,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology Letters","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s10529-026-03717-5","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
The isolate A85 was identified as Streptomyces enissocaesilis, and it might serve as a novel actinobacteria source for uricase production. The uricase was successfully produced and purified from this novel strain, demonstrating that it could be an efficient source for future applications. Characterization studies of the purified enzyme showed maximal activity at pH 9 and 37 °C. From the differential scanning calorimetry, a melting point of 55.6 °C was observed, which ensured that the purified uricase is thermostable. Overall, 32 fold purification with a 48% yield of uricase was recovered at the end of purification steps. Inhibitor studies revealed that the purified uricase is metalloprotease. Vmax and Km values were found to be 30.39 U/mL and 0.0212 mM, respectively. The molecular mass of the purified uricase was found to be around 32 KDa. Further MALDI-TOF MS analysis identified four peptides as uricase, with a significant score. The purified enzyme was classified as an uricase based on a comprehensive analysis that integrated its amino acid sequence and conserved regions. The secondary structure of the purified uricase using PDBsum revealed 1 beta sheet, 1 beta hairpin, 1 beta bulge, 2 strands, and 2 beta turns. The 3D structure of the purified uricase was predicted through homology modeling, and the structure was validated using Ramachandran plot analysis. Docking results revealed significant binding affinity of -4.3 kcal/mol, which indicates strong and stable interaction of uricase and uric acid substrate.
期刊介绍：
Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them.
All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included.
Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields.
The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories.
Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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