{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T04:58:07Z","timestamp":1784005087527,"version":"3.55.0"},"reference-count":55,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,6,13]],"date-time":"2023-06-13T00:00:00Z","timestamp":1686614400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012190","name":"Ministry of Science and Higher Education of the Russian Federation","doi-asserted-by":"publisher","award":["0287-2021-0020"],"award-info":[{"award-number":["0287-2021-0020"]}],"id":[{"id":"10.13039\/501100012190","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>A complex heterogeneous intracellular environment seems to affect enzymatic catalysis by changing the mobility of biomolecules, their stability, and their conformational states, as well as by facilitating or hindering continuously occurring interactions. The evaluation and description of the influence of the cytoplasmic matrix components on enzymatic activity are problems that remain unsolved. In this work, we aimed to determine the mechanisms of action of two-component media with cosolvents of various molecular sizes on the complex multi-stage bioluminescent reaction catalyzed by bacterial luciferase. Kinetic and structural effects of ethylene glycol, glycerol, sorbitol, glucose, sucrose, dextran, and polyethylene glycol on bacterial luciferase were studied using stopped-flow and fluorescence spectroscopy techniques and molecular dynamics simulations. We have found that diffusion limitations in the presence of cosolvents promote the stabilization of flavin substrate and peroxyflavin intermediate of the reaction, but do not provide any advantages in bioluminescence quantum yield, because substrate binding is slowed down as well. The catalytic constant of bacterial luciferase has been found to be viscosity-independent and correlated with parameters of water\u2013cosolvent interactions (Norrish constant, van der Waals interaction energies). Crowding agents, in contrast to low-molecular-weight cosolvents, had little effect on peroxyflavin intermediate decay and enzyme catalytic constant. We attributed specific kinetic effects to the preferential interaction of the cosolvents with enzyme surface and their penetration into the active site.<\/jats:p>","DOI":"10.3390\/life13061384","type":"journal-article","created":{"date-parts":[[2023,6,14]],"date-time":"2023-06-14T01:32:15Z","timestamp":1686706335000},"page":"1384","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["The Role of Cosolvent\u2013Water Interactions in Effects of the Media on Functionality of Enzymes: A Case Study of Photobacterium leiognathi Luciferase"],"prefix":"10.3390","volume":"13","author":[{"given":"Albert E.","family":"Lisitsa","sequence":"first","affiliation":[{"name":"Biophysics Department, Siberian Federal University, 660041 Krasnoyarsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lev A.","family":"Sukovatyi","sequence":"additional","affiliation":[{"name":"Biophysics Department, Siberian Federal University, 660041 Krasnoyarsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1883-8541","authenticated-orcid":false,"given":"Anna A.","family":"Deeva","sequence":"additional","affiliation":[{"name":"Biophysics Department, Siberian Federal University, 660041 Krasnoyarsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8537-4575","authenticated-orcid":false,"given":"Dmitry V.","family":"Gulnov","sequence":"additional","affiliation":[{"name":"Biophysics Department, Siberian Federal University, 660041 Krasnoyarsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2560-3575","authenticated-orcid":false,"given":"Elena N.","family":"Esimbekova","sequence":"additional","affiliation":[{"name":"Biophysics Department, Siberian Federal University, 660041 Krasnoyarsk, Russia"},{"name":"Institute of Biophysics, Siberian Branch of Russian Academy of Sciences, 660036 Krasnoyarsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valentina A.","family":"Kratasyuk","sequence":"additional","affiliation":[{"name":"Biophysics Department, Siberian Federal University, 660041 Krasnoyarsk, Russia"},{"name":"Institute of Biophysics, Siberian Branch of Russian Academy of Sciences, 660036 Krasnoyarsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1725-8625","authenticated-orcid":false,"given":"Elena V.","family":"Nemtseva","sequence":"additional","affiliation":[{"name":"Biophysics Department, Siberian Federal University, 660041 Krasnoyarsk, Russia"},{"name":"Institute of Biophysics, Siberian Branch of Russian Academy of Sciences, 660036 Krasnoyarsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3234","DOI":"10.1016\/S0006-3495(99)77154-7","article-title":"Crowding effects on EcoRV kinetics and binding","volume":"77","author":"Wenner","year":"1999","journal-title":"Biophys. 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