{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,17]],"date-time":"2026-01-17T19:27:33Z","timestamp":1768678053808,"version":"3.49.0"},"reference-count":35,"publisher":"Bentham Science Publishers Ltd.","issue":"6","content-domain":{"domain":["eurekaselect.com"],"crossmark-restriction":true},"short-container-title":["PPL"],"published-print":{"date-parts":[[2021,6,25]]},"abstract":"<jats:sec><jats:title>Background:<\/jats:title><jats:p>Fibrinolytic protease from Euphausia superba (EFP) was isolated.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective:<\/jats:title><jats:p>Biochemical distinctions, regulation of the catalytic function, and the key residues of EFP were investigated.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods:<\/jats:title><jats:p>The serial inhibition kinetic evaluations coupled with measurements of fluorescence spectra in the presence of 4-(2-aminoethyl) benzene sulfonyl fluoride hydrochloride (AEBSF) was conducted. The computational molecular dynamics (MD) simulations were also applied for a comparative study.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results:<\/jats:title><jats:p>The enzyme behaved as a monomeric protein with a molecular mass of about 28.6 kD with K<jats:sub>m<\/jats:sub><jats:sup>BApNA<\/jats:sup>= 0.629 \u00b1 0.02 mM and k<jats:sub>cat<\/jats:sub>\/K<jats:sub>m<\/jats:sub><jats:sup>BApNA<\/jats:sup>= 7.08 s-1\/mM. The real-time interval measurements revealed that the inactivation was a first-order reaction, with the kinetic processes shifting from a monophase to a biphase. Measurements of fluorescence spectra showed that serine residue modification by AEBSF directly caused conspicuous changes of the tertiary structures and exposed hydrophobic surfaces. Some osmolytes were applied to find protective roles. These results confirmed that the active region of EFP is more flexible than the overall enzyme molecule and serine, as the key residue, is associated with the regional unfolding of EFP in addition to its catalytic role. The MD simulations were supportive to the kinetics data.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion:<\/jats:title><jats:p>Our study indicated that EFP has an essential serine residue for its catalyst function and associated folding behaviors. Also, the functional role of osmolytes such as proline and glycine that may play a role in defense mechanisms from environmental adaptation in a krill\u2019s body was suggested.<\/jats:p><\/jats:sec>","DOI":"10.2174\/0929866527666201112123714","type":"journal-article","created":{"date-parts":[[2020,11,13]],"date-time":"2020-11-13T04:31:53Z","timestamp":1605241913000},"page":"651-664","update-policy":"https:\/\/doi.org\/10.2174\/bsp_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity"],"prefix":"10.2174","volume":"28","author":[{"given":"Guo-Ying","family":"Qian","sequence":"first","affiliation":[{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gyutae","family":"Lim","sequence":"additional","affiliation":[{"name":"Genome Editing Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Gwahak-ro, Yuseonggu, Daejeon, 34141, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shang-Jun","family":"Yin","sequence":"additional","affiliation":[{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun-Mo","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Sungkyunkwan University School of Medicine, Samsung Medical Center, Seoul 135-710, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinhyuk","family":"Lee","sequence":"additional","affiliation":[{"name":"Genome Editing Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Gwahak-ro, Yuseonggu, Daejeon, 34141, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong-Doo","family":"Park","sequence":"additional","affiliation":[{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"965","reference":[{"key":"ref=1","doi-asserted-by":"publisher","first-page":"426","DOI":"10.1016\/j.ijbiomac.2014.03.044","volume":"67","author":"Si Y.X.","year":"2014","unstructured":"Si Y.X.; Song J.J.; Fang N.Y.; Wang W.; Wang Z.J.; Yang J.M.; Qian G.Y.; Yin S.J.; Park Y.D.; Purification, characterization, and unfolding studies of arginine kinase from Antarctic krill. Int J Biol Macromol 2014,67,426-432","journal-title":"Int J Biol Macromol"},{"key":"ref=2","doi-asserted-by":"publisher","first-page":"3888","DOI":"10.1007\/s12010-014-0802-9","volume":"172","author":"Si Y.X.","year":"2014","unstructured":"Si Y.X.; Fang N.Y.; Wang W.; Wang Z.J.; Yang J.M.; Qian G.Y.; Yin S.J.; Park Y.D.; Folding studies of arginine kinase from Euphausia superba using denaturants. Appl Biochem Biotechnol 2014,172(8),3888-3901","journal-title":"Appl Biochem Biotechnol"},{"key":"ref=3","doi-asserted-by":"publisher","first-page":"266","DOI":"10.1016\/j.ijbiomac.2014.07.001","volume":"70","author":"Wang Z.J.","year":"2014","unstructured":"Wang Z.J.; Lee J.; Si Y.X.; Wang W.; Yang J.M.; Yin S.J.; Qian G.Y.; Park Y.D.; A folding study of Antarctic krill (Euphausia superba) alkaline phosphatase using denaturants. Int J Biol Macromol 2014,70,266-274","journal-title":"Int J Biol Macromol"},{"key":"ref=4","doi-asserted-by":"publisher","first-page":"936","DOI":"10.1016\/j.procbio.2014.03.019","volume":"49","author":"Fang N.Y.","year":"2014","unstructured":"Fang N.Y.; Lee J.; Yin S.J.; Wang W.; Wang Z.J.; Yang J.M.; Qian G.Y.; Si Y.X.; Park Y.D.; Effects of osmolytes on arginine kinase from Euphausia superba: a study on thermal denaturation and aggregation. Process Biochem 2014,49,936-947","journal-title":"Process Biochem"},{"key":"ref=5","doi-asserted-by":"publisher","first-page":"821","DOI":"10.1016\/j.procbio.2014.02.004","volume":"49","author":"Jin Q.X.","year":"2014","unstructured":"Jin Q.X.; Yin S.J.; Wang W.; Wang Z.J.; Yang J.M.; Qian G.Y.; Si Y.X.; Park Y.D.; The effect of Zn on Euphausia superba arginine kinase: unfolding and aggregation studies. Process Biochem 2014,49,821-829","journal-title":"Process Biochem"},{"key":"ref=6","doi-asserted-by":"publisher","first-page":"395","DOI":"10.1016\/j.procbio.2014.12.010","volume":"50","author":"Cheng J.G.","year":"2015","unstructured":"Cheng J.G.; Si Y.X.; Lee J.; Zhao F.; Yang J.M.; Qian G.Y.; Yin S.J.; Park Y.D.; The effect of Cu on arginine kinase from Euphausia superba: a computational simulation integrating unfolding and aggregation studies. Process Biochem 2015,50,395-405","journal-title":"Process Biochem"},{"key":"ref=7","first-page":"85","volume":"33","author":"Turkiewicz M.","year":"1986","unstructured":"Turkiewicz M.; Galas E.; Kalinowska H.; Romanowska I.; Zieli\u0144ska M.; Purification and characterization of a proteinase from Euphausia superba Dana (Antarctic krill). Acta Biochim Pol 1986,33(2),85-99","journal-title":"Acta Biochim Pol"},{"key":"ref=8","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1016\/0305-0491(91)90056-J","volume":"99","author":"Turkiewicz M.","year":"1991","unstructured":"Turkiewicz M.; Galas E.; Kalinowska H.; Collagenolytic serine proteinase from Euphausia superba Dana (Antarctic krill). Comp Biochem Physiol B 1991,99(2),359-371","journal-title":"Comp Biochem Physiol B"},{"key":"ref=9","first-page":"79","volume":"38","author":"Turkiewicz M.","year":"1991","unstructured":"Turkiewicz M.; Kalinowska H.; Galas E.; An endo-(1----3)-beta-glucanase and a collagenolytic serine proteinase from Euphausia superba Dana (Antarctic krill). Acta Biochim Pol 1991,38(1),79-85","journal-title":"Acta Biochim Pol"},{"key":"ref=10","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1016\/S1046-5928(02)00519-3","volume":"26","author":"Sj\u00f6dahl J.","year":"2002","unstructured":"Sj\u00f6dahl J.; Emmer A.; Vincent J.; Roeraade J.; Characterization of proteinases from Antarctic krill (Euphausia superba). Protein Expr Purif 2002,26(1),153-161","journal-title":"Protein Expr Purif"},{"key":"ref=11","first-page":"99","volume":"20","author":"Hellgren K.","year":"2009","unstructured":"Hellgren K.; Assessment of Krillase chewing gum for the reduction of gingivitis and dental plaque. J Clin Dent 2009,20(3),99-102","journal-title":"J Clin Dent"},{"key":"ref=12","doi-asserted-by":"publisher","first-page":"316","DOI":"10.1034\/j.1600-0722.2001.00099.x","volume":"109","author":"Berg C.H.","year":"2001","unstructured":"Berg C.H.; Kalfas S.; Malmsten M.; Arnebrant T.; Proteolytic degradation of oral biofilms in vitro and in vivo: potential of proteases originating from Euphausia superba for plaque control. Eur J Oral Sci 2001,109(5),316-324","journal-title":"Eur J Oral Sci"},{"key":"ref=13","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1046\/j.1432-1327.2001.01857.x","volume":"268","author":"Benjamin D.C.","year":"2001","unstructured":"Benjamin D.C.; Kristj\u00e1nsd\u00f3ttir S.; Gudmundsd\u00f3ttir A.; Increasing the thermal stability of euphauserase. A cold-active and multifunctional serine protease from Antarctic krill. Eur J Biochem 2001,268(1),127-131","journal-title":"Eur J Biochem"},{"key":"ref=14","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1007\/BF02118628","volume":"42","author":"Hellgren L.","year":"1986","unstructured":"Hellgren L.; Mohr V.; Vincent J.; Proteases of Antarctic krill--a new system for effective enzymatic debridement of necrotic ulcerations. Experientia 1986,42(4),403-404","journal-title":"Experientia"},{"key":"ref=15","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1007\/BF00426587","volume":"281","author":"Anheller J.E.","year":"1989","unstructured":"Anheller J.E.; Hellgren L.; Karlstam B.; Vincent J.; Biochemical and biological profile of a new enzyme preparation from Antarctic krill (E. superba) suitable for debridement of ulcerative lesions. Arch Dermatol Res 1989,281(2),105-110","journal-title":"Arch Dermatol Res"},{"key":"ref=16","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1097\/00003226-199911000-00014","volume":"18","author":"Sangwan V.S.","year":"1999","unstructured":"Sangwan V.S.; Akpek E.K.; Voo I.; Zhao T.; Pinar V.; Yang J.; Christen W.; Baltatzis S.; Wild R.; Foster C.S.; Krill protease effects on wound healing after corneal alkali burn. Cornea 1999,18(6),707-711","journal-title":"Cornea"},{"key":"ref=17","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1016\/S0896-6273(00)80290-1","volume":"17","author":"Citron M.","year":"1996","unstructured":"Citron M.; Diehl T.S.; Capell A.; Haass C.; Teplow D.B.; Selkoe D.J.; Inhibition of amyloid beta-protein production in neural cells by the serine protease inhibitor AEBSF. Neuron 1996,17(1),171-179","journal-title":"Neuron"},{"key":"ref=18","doi-asserted-by":"publisher","first-page":"4293","DOI":"10.1021\/acs.analchem.7b05316","volume":"90","author":"Huang J.L.","year":"2018","unstructured":"Huang J.L.; Nagy A.; Ivleva V.B.; Blackstock D.; Arnold F.; Cai C.X.; Hydrolysis-kinetic study of AEBSF, a protease inhibitor used during cell-culture processing of the HIV-1 broadly neutralizing antibody CAP256-VRC25.26. Anal Chem 2018,90(7),4293-4296","journal-title":"Anal Chem"},{"key":"ref=19","first-page":"e25180735","volume":"91","author":"Souto X.M.","year":"2019","unstructured":"Souto X.M.; Branquinha M.H.; Santos A.L.S.; Chymotrypsin- and trypsin-like activities secreted by the multidrug-resistant yeasts forming the Candida haemulonii complex An Acad Bras Cienc 2019,91,e25180735","journal-title":"An Acad Bras Cienc"},{"key":"ref=20","doi-asserted-by":"crossref","first-page":"1060","DOI":"10.1021\/acs.bioconjchem.7b00717","volume":"29","author":"Lim\u00f3n D.","year":"2018","unstructured":"Lim\u00f3n D.; F\u00e1brega M.J.; Calpena A.C.; Badia J.; Baldom\u00e0 L.; P\u00e9rez-Garc\u00eda L.; Multifunctional serine protease inhibitor-coated water-soluble gold nanoparticles as a novel targeted approach for the treatment of inflammatory skin diseases Bioconjug Chem 2018,29,1060-1072","journal-title":"Bioconjug Chem"},{"key":"ref=21","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1006\/bbrc.1998.9760","volume":"253","author":"Chu T.M.","year":"1998","unstructured":"Chu T.M.; Kawinski E.; Plasmin, substilisin-like endoproteases, tissue plasminogen activator, and urokinase plasminogen activator are involved in activation of latent TGF-beta 1 in human seminal plasma. Biochem Biophys Res Commun 1998,253(1),128-134","journal-title":"Biochem Biophys Res Commun"},{"key":"ref=22","doi-asserted-by":"publisher","first-page":"157","DOI":"10.1186\/s12985-017-0824-3","volume":"14","author":"Van der Gucht W.","year":"2017","unstructured":"Van der Gucht W.; Leemans A.; De Schryver M.; Heykers A.; Caljon G.; Maes L.; Cos P.; Delputte P.L.; Respiratory syncytial virus (RSV) entry is inhibited by serine protease inhibitor AEBSF when present during an early stage of infection. Virol J 2017,14(1),157","journal-title":"Virol J"},{"key":"ref=23","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2019.197672","volume":"271","author":"Sreelatha L.","year":"2019","unstructured":"Sreelatha L.; Malakar S.; Songprakhon P.; Morchang A.; Srisawat C.; Noisakran S.; Yenchitosomanus P.T.; Limjindaporn T.; Serine protease inhibitor AEBSF reduces dengue virus infection via decreased cholesterol synthesis. Virus Res 2019,271","journal-title":"Virus Res"},{"key":"ref=24","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1023\/A:1016383212244","volume":"10","author":"Salamanca M.H.","year":"2001","unstructured":"Salamanca M.H.; Barr\u00eda C.; Asenjo J.A.; Andrews B.A.; Isolation, purification and preliminary characterization of cryophilic proteases of marine origin. Bioseparation 2001,10(4-5),237-241","journal-title":"Bioseparation"},{"key":"ref=25","doi-asserted-by":"publisher","first-page":"7573","DOI":"10.1021\/bi953067l","volume":"35","author":"Tams J.W.","year":"1996","unstructured":"Tams J.W.; Welinder K.G.; Unfolding and refolding of Coprinus cinereus peroxidase at high pH, in urea, and at high temperature. Effect of organic and ionic additives on these processes. Biochemistry 1996,35(23),7573-7579","journal-title":"Biochemistry"},{"key":"ref=26","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1016\/j.ijbiomac.2018.05.165","volume":"117","author":"Wang Z.J.","year":"2018","unstructured":"Wang Z.J.; Ma W.; Yang J.M.; Kang Y.; Park Y.D.; Effects of Cu on alkaline phosphatase from Macrobrachium rosenbergii. Int J Biol Macromol 2018,117,116-123","journal-title":"Int J Biol Macromol"},{"key":"ref=27","doi-asserted-by":"publisher","first-page":"667","DOI":"10.1016\/j.ijbiomac.2018.02.011","volume":"112","author":"Wang Z.J.","year":"2018","unstructured":"Wang Z.J.; Zheng L.; Yang J.M.; Kang Y.; Park Y.D.; Proteomic analyses for profiling regulated proteins\/enzymes by Fucus vesiculosus fucoidan in B16 melanoma cells: A combination of enzyme kinetics functional study. Int J Biol Macromol 2018,112,667-674","journal-title":"Int J Biol Macromol"},{"key":"ref=28","doi-asserted-by":"publisher","first-page":"W296","DOI":"10.1093\/nar\/gky427","volume":"46","author":"Waterhouse A.","year":"2018","unstructured":"Waterhouse A.; Bertoni M.; Bienert S.; Studer G.; Tauriello G.; Gumienny R.; Heer F.T.; de Beer T.A.P.; Rempfer C.; Bordoli L.; Lepore R.; Schwede T.; SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 2018,46(W1),W296-W303","journal-title":"Nucleic Acids Res"},{"key":"ref=29","doi-asserted-by":"publisher","first-page":"D48","DOI":"10.1093\/nar\/gkr1202","volume":"40","author":"Benson D.A.","year":"2012","unstructured":"Benson D.A.; Karsch-Mizrachi I.; Clark K.; Lipman D.J.; Ostell J.; Sayers E.W.; GenBank. Nucleic Acids Res 2012,40(Database issue),D48-D53","journal-title":"Nucleic Acids Res"},{"key":"ref=30","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1038\/nmeth.1818","volume":"9","author":"Remmert M.","year":"2011","unstructured":"Remmert M.; Biegert A.; Hauser A.; S\u00f6ding J.; HHblits: lightning-fast iterative protein sequence searching by HMM-HMM alignment. Nat Methods 2011,9(2),173-175","journal-title":"Nat Methods"},{"key":"ref=31","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/0263-7855(96)00018-5","volume":"14","author":"Humphrey W.","year":"1996","unstructured":"Humphrey W.; Dalke A.; Schulten K.; VMD: visual molecular dynamics J Mol Graph 1996,14,33-38","journal-title":"J Mol Graph"},{"key":"ref=32","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1002\/jcc.21334","volume":"31","author":"Trott O.","year":"2010","unstructured":"Trott O.; Olson A.J.; AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem 2010,31(2),455-461","journal-title":"J Comput Chem"},{"key":"ref=33","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/j.softx.2015.06.001","volume":"1-2","author":"Abraham M.J.","year":"2015","unstructured":"Abraham M.J.; Murtola T.; Schulz R.; P\u00e1ll S.; Smith J.C.; Hess B.; Lindahl E.; GROMACS: high performance molecular simulations through multi-level parallelism from laptops to supercomputers. SoftwareX 2015,1-2,19-25","journal-title":"SoftwareX"},{"key":"ref=34","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1002\/jcc.21367","volume":"31","author":"Vanommeslaeghe K.","year":"2010","unstructured":"Vanommeslaeghe K.; Hatcher E.; Acharya C.; Kundu S.; Zhong S.; Shim J.; Darian E.; Guvench O.; Lopes P.; Vorobyov I.; Mackerell A.D.; CHARMM general force field: a force field for drug-like molecules compatible with the CHARMM all-atom additive biological force fields. J Comput Chem 2010,31(4),671-690","journal-title":"J Comput Chem"},{"key":"ref=35","doi-asserted-by":"publisher","first-page":"1605","DOI":"10.1002\/jcc.20084","volume":"25","author":"Pettersen E.F.","year":"2004","unstructured":"Pettersen E.F.; Goddard T.D.; Huang C.C.; Couch G.S.; Greenblatt D.M.; Meng E.C.; Ferrin T.E.; UCSF Chimera a visualization system for exploratory research and analysis. J Comput Chem 2004,25(13),1605-1612","journal-title":"J Comput Chem"}],"container-title":["Protein &amp; Peptide Letters"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/eurekaselect.com\/article\/download\/187909","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.eurekaselect.com\/187909\/article","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/eurekaselect.com\/article\/download\/187909","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,8,17]],"date-time":"2024-08-17T10:17:54Z","timestamp":1723889874000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.eurekaselect.com\/187909\/article"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,25]]},"references-count":35,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2021,6,25]]}},"alternative-id":["LiveAll1"],"URL":"https:\/\/doi.org\/10.2174\/0929866527666201112123714","relation":{},"ISSN":["0929-8665"],"issn-type":[{"value":"0929-8665","type":"print"}],"subject":[],"published":{"date-parts":[[2021,6,25]]},"assertion":[{"value":"Peer Reviewed","order":0,"name":"review_status","label":"Review Status","group":{"name":"peer_review_details","label":"Peer Review Details"}},{"value":"Single blind","order":1,"name":"review_process","label":"Review Process","group":{"name":"peer_review_details","label":"Peer Review Details"}},{"value":"Checked with iThenticate","order":0,"name":"screening_status","label":"Screening Status","group":{"name":"plagiarism_screening","label":"Plagiarism Screening"}},{"value":"2020-05-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-09-27","order":1,"name":"revised","label":"Revised","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-10-02","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-06-25","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}