From Secretion in Pichia pastoris to Application in Apple Juice Processing: Exo-Polygalacturonase from Sporothrix schenckii 1099-18

dc.authoridKARATAS, Ersin/0000-0001-6848-7618
dc.authoridTulek, Ahmet/0000-0003-1079-7837
dc.authoridAKTAS, Fatih/0000-0002-2031-298X
dc.authoridbinay, baris/0000-0002-6190-6549
dc.authorwosidKARATAS, Ersin/G-1852-2016
dc.authorwosidTulek, Ahmet/S-8916-2018
dc.authorwosidBinay, Baris/ABB-7968-2021
dc.contributor.authorKaratas, Ersin
dc.contributor.authorTulek, Ahmet
dc.contributor.authorCakar, Mehmet Mervan
dc.contributor.authorTamturk, Faruk
dc.contributor.authorAktas, Fatih
dc.contributor.authorBinay, Baris
dc.date.accessioned2021-12-01T18:49:03Z
dc.date.available2021-12-01T18:49:03Z
dc.date.issued2021
dc.department[Belirlenecek]en_US
dc.description.abstractBackground: Polygalacturonases are a group of enzymes under pectinolytic enzymes related to enzymes that hydrolyse pectic substances. Polygalacturonases have been used in various industrial applications such as fruit juice clarification, retting of plant fibers, wastewater treatment drinks fermentation, and oil extraction. Objectives: The study was evaluated at the heterologous expression, purification, biochemical characterization, computational modeling, and performance in apple juice clarification of a new exo-polygalacturonase from Sporothrix schenckii 1099-18 (SsExo-PG) in Pichia pastoris. Methods: Recombinant DNA technology was used in this study. Two different pPIC9K plasmids were constructed with native signal sequence-ssexo-pg and alpha signal sequence-ssexo-pg separately. Protein expression and purification performed after plasmids transformed into the Pichia pastoris. Biochemical and structural analyses were performed by using pure SsExo-PG. Results: The purification of SsExo-PG was achieved using a Ni-NTA chromatography system. The enzyme was found to have a molecular mass of approximately 52 kDa. SsExo-PG presented as stable at a wide range of temperature and pH values, and to be more storage stable than other commercial pectinolytic enzyme mixtures. Structural analysis revealed that the catalytic residues of SsEx-o-PG are somewhat similar to other Exo-PGs. The K-M and k(cat) values for the degradation of polygalacturonic acid (PGA) by the purified enzyme were found to be 0.5868 mu M and 179 s(-1), respectively. Cu2+ was found to enhance SsExo-PG activity while Ag2+ and Fe2+ almost completely inhibited enzyme activity. The enzyme reduced turbidity up to 80% thus enhanced the clarification of apple juice. SsExo-PG showed promising performance when compared with other commercial pectinolytic enzyme mixtures. Conclusion: The clarification potential of SsExo-PG was revealed by comparing it with commercial pectinolytic enzymes. The following parameters of the process of apple juice clarification processes showed that SsExo-PG is highly stable and has a novel performance.en_US
dc.description.sponsorshipDuzce University Scientific Research Projects Department (DUBAP)Duzce University [2020.06.02.1092]en_US
dc.description.sponsorshipThis work was fully supported by a grant from the Duzce University Scientific Research Projects Department (DUBAP) (Grant number: 2020.06.02.1092).en_US
dc.identifier.doi10.2174/1871530321666210106110400
dc.identifier.endpage830en_US
dc.identifier.issn0929-8665
dc.identifier.issn1875-5305
dc.identifier.issue7en_US
dc.identifier.pmid33413052en_US
dc.identifier.scopus2-s2.0-85115040276en_US
dc.identifier.scopusqualityQ3en_US
dc.identifier.startpage817en_US
dc.identifier.urihttps://doi.org/10.2174/1871530321666210106110400
dc.identifier.urihttps://hdl.handle.net/20.500.12684/10655
dc.identifier.volume28en_US
dc.identifier.wosWOS:000683897300001en_US
dc.identifier.wosqualityQ4en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakPubMeden_US
dc.indekslendigikaynakScopusen_US
dc.language.isoenen_US
dc.publisherBentham Science Publ Ltden_US
dc.relation.ispartofProtein And Peptide Lettersen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectSporothrix schenckii 1099-18en_US
dc.subjectExo-polygalacturonaseen_US
dc.subjectheterologous expressionen_US
dc.subjectPichia pastorisen_US
dc.subjectapple juice clarifi-cationen_US
dc.subjectstructural modellingen_US
dc.subjectEndo-Polygalacturonaseen_US
dc.subjectBiochemical-Characterizationen_US
dc.subjectCrystal-Structureen_US
dc.subjectGlycan Analysisen_US
dc.subjectAspergillusen_US
dc.subjectPurificationen_US
dc.subjectEnzymesen_US
dc.subjectExopolygalacturonaseen_US
dc.subjectExpressionen_US
dc.subjectEndopolygalacturonaseen_US
dc.titleFrom Secretion in Pichia pastoris to Application in Apple Juice Processing: Exo-Polygalacturonase from Sporothrix schenckii 1099-18en_US
dc.typeArticleen_US

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