{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T00:43:46Z","timestamp":1772585026688,"version":"3.50.1"},"reference-count":33,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2014,1,1]],"date-time":"2014-01-01T00:00:00Z","timestamp":1388534400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2014,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Typical plant aspartic protease zymogens comprise a characteristic and plant-specific insert (PSI). PSI domains can interact with membranes, and a role as a defensive weapon against pathogens has been proposed. However, the potential of PSIs as antimicrobial agents has not been fully investigated and explored yet due to problems in producing sufficient amounts of these domains in bacteria. Here, we report the development of an expression platform for the production of the PSI domain of cirsin in the generally regarded as safe (GRAS) yeast\n            <jats:named-content content-type=\"genus-species\">Kluyveromyces lactis<\/jats:named-content>\n            . We successfully generated\n            <jats:named-content content-type=\"genus-species\">K. lactis<\/jats:named-content>\n            transformants expressing and secreting significant amounts of correctly processed and glycosylated PSI, as well as its nonglycosylated mutant. A purification protocol with protein yields of \u223c4.0 mg\/liter was established for both wild-type and nonglycosylated PSIs, which represents the highest reported yield for a nontagged PSI domain. Subsequent bioactivity assays targeting phytopathogenic fungi indicated that the PSI of cirsin is produced in a biologically active form in\n            <jats:named-content content-type=\"genus-species\">K. lactis<\/jats:named-content>\n            and provided clear evidence for its antifungal activity. This yeast expression system thereby emerges as a promising production platform for further exploring the biotechnological potential of these plant saposin-like proteins.\n          <\/jats:p>","DOI":"10.1128\/aem.03151-13","type":"journal-article","created":{"date-parts":[[2013,10,12]],"date-time":"2013-10-12T01:21:53Z","timestamp":1381540913000},"page":"86-96","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":17,"title":["Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin"],"prefix":"10.1128","volume":"80","author":[{"given":"Pedro","family":"Curto","sequence":"first","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Coimbra, Portugal"},{"name":"Biocant, Biotechnology Innovation Center, Cantanhede, Portugal"}]},{"given":"Daniela","family":"Lufrano","sequence":"additional","affiliation":[{"name":"Servei de Prote\u00f2mica i Biologia Estructural, Universitat Aut\u00f2noma de Barcelona, Campus Universitari, Bellaterra, Cerdanyola del Vall\u00e8s, Barcelona, Spain"}]},{"given":"C\u00e1tia","family":"Pinto","sequence":"additional","affiliation":[{"name":"Biocant, Biotechnology Innovation Center, Cantanhede, Portugal"}]},{"given":"Val\u00e9ria","family":"Cust\u00f3dio","sequence":"additional","affiliation":[{"name":"Biocant, Biotechnology Innovation Center, Cantanhede, Portugal"}]},{"given":"Ana Catarina","family":"Gomes","sequence":"additional","affiliation":[{"name":"Biocant, Biotechnology Innovation Center, Cantanhede, Portugal"}]},{"given":"Sebasti\u00e1n A.","family":"Trejo","sequence":"additional","affiliation":[{"name":"Servei de Prote\u00f2mica i Biologia Estructural, Universitat Aut\u00f2noma de Barcelona, Campus Universitari, Bellaterra, Cerdanyola del Vall\u00e8s, Barcelona, Spain"}]},{"given":"Laura","family":"Bak\u00e1s","sequence":"additional","affiliation":[{"name":"Laboratorio 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