Приказ основних података о документу

dc.creatorRašković, Brankica
dc.creatorVatić, Saša
dc.creatorAnđelković, Boban D.
dc.creatorBlagojević, Vladimir A.
dc.creatorPolović, Natalija
dc.date.accessioned2018-11-22T00:35:21Z
dc.date.available2018-11-22T00:35:21Z
dc.date.issued2016
dc.identifier.issn1369-703X
dc.identifier.urihttps://cherry.chem.bg.ac.rs/handle/123456789/2024
dc.description.abstractTrypsin is a serine protease with widespread applications, including protein sequencing and typsin mass fingerprinting. In the present study, the storage of trypsin in acidic conditions significantly affected the recovery of activity (40%) after 7 freeze-thaw cycles. Further, trypsin lost parts of its native secondary structure elements, which resulted in a 10% increase in beta-sheet content (band maximum detected at a frequency of 1634 cm in the Fourier transform infrared (FT-IR) spectrum) indicative of freezing-induced denaturation of the protein. The cold storage of trypsin in ammonium bicarbonate (pH 8.2) with the addition of ayoprotectants, such as glycerol or lysine, led to protein stabilization (complete secondary structure content preservation was detected by FT-IR), higher activity recovery ( gt 90%) and modest autolysis ( lt 10%). High activity recovery ( gt 90%) was also detected with the addition of propylene glycol and polyethylene glycol, saccharides and arginine. Nevertheless, trypsin stored at pH 8.2 with the addition of glycerol or lysine was as efficient as untreated trypsin in the trypsin mass fingerprinting analysis of BSA, suggesting that the cold storage of trypsin in slightly alkaline conditions with the addition of cryoprotectants could prolong its shelf life. (C) 2015 Elsevier B.V. All rights reserved.en
dc.publisherElsevier Science Bv, Amsterdam
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172049/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172057/RS//
dc.rightsrestrictedAccess
dc.sourceBiochemical Engineering Journal
dc.subjectEnzyme activityen
dc.subjectCold stabilityen
dc.subjectProtein recoveryen
dc.subjectProtein denaturationen
dc.subjectProteolysisen
dc.subjectProteomicsen
dc.titleOptimizing storage conditions to prevent cold denaturation of trypsin for sequencing and to prolong its shelf lifeen
dc.typearticle
dc.rights.licenseARR
dcterms.abstractAнделковиц, Бобан; Ватиц, Саса; Половић, Наталија; Благојевиц, Владимир; Расковиц, Бранкица;
dc.citation.volume105
dc.citation.spage168
dc.citation.epage176
dc.identifier.wos000367776300019
dc.identifier.doi10.1016/j.bej.2015.09.018
dc.citation.other105: 168-176
dc.citation.rankM21
dc.description.otherSupplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3586]
dc.type.versionpublishedVersionen
dc.identifier.scopus2-s2.0-84942792521


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Приказ основних података о документу