{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T17:57:04Z","timestamp":1785520624643,"version":"3.56.0"},"reference-count":33,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T00:00:00Z","timestamp":1712707200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Russian Science Foundation","award":["22-74-10100"],"award-info":[{"award-number":["22-74-10100"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>This study investigated the effect of polycationic and uncharged polymers (and oligomers) on the catalytic parameters and thermostability of L-asparaginase from Thermococcus sibiricus (TsA). This enzyme has potential applications in the food industry to decrease the formation of carcinogenic acrylamide during the processing of carbohydrate-containing products. Conjugation with the polyamines polyethylenimine and spermine (PEI and Spm) or polyethylene glycol (PEG) did not significantly affect the secondary structure of the enzyme. PEG contributes to the stabilization of the dimeric form of TsA, as shown by HPLC. Furthermore, neither polyamines nor PEG significantly affected the binding of the L-Asn substrate to TsA. The conjugates showed greater maximum activity at pH 7.5 and 85 \u00b0C, 10\u201350% more than for native TsA. The pH optima for both TsA-PEI and TsA-Spm conjugates were shifted to lower pH ranges from pH 10 (for the native enzyme) to pH 8.0. Additionally, the TsA-Spm conjugate exhibited the highest activity at pH 6.5\u20139.0 among all the samples. Furthermore, the temperature optimum for activity at pH 7.5 shifted from 90\u201395 \u00b0C to 80\u201385 \u00b0C for the conjugates. The thermal inactivation mechanism of TsA-PEG appeared to change, and no aggregation was observed in contrast to that of the native enzyme. This was visually confirmed and supported by the analysis of the CD spectra, which remained almost unchanged after heating the conjugate solution. These results suggest that TsA-PEG may be a more stable form of TsA, making it a potentially more suitable option for industrial use.<\/jats:p>","DOI":"10.3390\/ijms25084174","type":"journal-article","created":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T03:07:48Z","timestamp":1712718468000},"page":"4174","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["L-Asparaginase Conjugates from the Hyperthermophilic Archaea Thermococcus sibiricus with Improved Biocatalytic Properties"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-4405-725X","authenticated-orcid":false,"given":"Natalia V.","family":"Dobryakova","sequence":"first","affiliation":[{"name":"Group of Fungal Genetic Engineering, Federal Research Center \u201cFundamentals of Biotechnology of the Russian Academy of Sciences\u201d, 117312 Moscow, Russia"},{"name":"Chemical Faculty, Lomonosov Moscow State University, Leninskie Gory St. 1, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria V.","family":"Dumina","sequence":"additional","affiliation":[{"name":"Group of Fungal Genetic Engineering, Federal Research Center \u201cFundamentals of Biotechnology of the Russian Academy of Sciences\u201d, 117312 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8672-8638","authenticated-orcid":false,"given":"Alexander A.","family":"Zhgun","sequence":"additional","affiliation":[{"name":"Group of Fungal Genetic Engineering, Federal Research Center \u201cFundamentals of Biotechnology of the Russian Academy of Sciences\u201d, 117312 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-2726-3632","authenticated-orcid":false,"given":"Marina V.","family":"Pokrovskaya","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Biotechnology, Institute of Biomedical Chemistry, 119121 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4248-765X","authenticated-orcid":false,"given":"Svetlana S.","family":"Aleksandrova","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Biotechnology, Institute of Biomedical Chemistry, 119121 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4753-7588","authenticated-orcid":false,"given":"Dmitry D.","family":"Zhdanov","sequence":"additional","affiliation":[{"name":"Group of Fungal Genetic Engineering, Federal Research Center \u201cFundamentals of Biotechnology of the Russian Academy of Sciences\u201d, 117312 Moscow, Russia"},{"name":"Laboratory of Medical Biotechnology, Institute of Biomedical Chemistry, 119121 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9761-7757","authenticated-orcid":false,"given":"Elena V.","family":"Kudryashova","sequence":"additional","affiliation":[{"name":"Group of Fungal Genetic Engineering, Federal Research Center \u201cFundamentals of Biotechnology of the Russian Academy of Sciences\u201d, 117312 Moscow, Russia"},{"name":"Chemical Faculty, Lomonosov Moscow State University, Leninskie Gory St. 1, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Jia, R., Wan, X., Geng, X., Xue, D., Xie, Z., and Chen, C. 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