{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:11:54Z","timestamp":1785409914131,"version":"3.56.0"},"reference-count":62,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100020399","name":"Yayasan Penyelidikan Antartika Sultan Mizan","doi-asserted-by":"crossref","award":["LPS2309"],"award-info":[{"award-number":["LPS2309"]}],"id":[{"id":"10.13039\/501100020399","id-type":"DOI","asserted-by":"crossref"}]},{"award":["LPS2309"],"award-info":[{"award-number":["LPS2309"]}],"id":[{"id":"https:\/\/ror.org\/02v2qzh60","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Antarctic microorganisms experience persistent subzero temperatures and repeated freeze\u2013thaw cycles that require specialized mechanisms for survival. Although transcriptomic studies have identified numerous cold-responsive genes, many remain annotated as hypothetical proteins with unknown functions. In this study, we investigated PL002-1792, a strongly upregulated hypothetical protein from Antarctic Flavobacterium sp. PL002 identified under severe cold stress (\u221220 \u00b0C versus \u22126 \u00b0C; log2 fold change = 5.61, adjusted p = 1.89 \u00d7 10\u2212138). Sequence analysis revealed a 402-amino-acid protein containing a predicted Sec\/SPII lipoprotein signal peptide. AlphaFold3 prediction generated a high-confidence structural model (pTM = 0.96) with an elongated \u03b2-sheet-rich architecture resembling bacterial ice-binding proteins. Comparative analysis with the ice-binding protein from Flavobacterium frigoris (FfIBP) identified conserved glycine-rich and TXT-like motifs associated with putative ice-binding surfaces. Recombinant PL002-1792 was expressed in Escherichia coli, recovered from inclusion bodies, and successfully refolded into a predominantly \u03b2-sheet-rich conformation as confirmed by circular dichroism spectroscopy. Functional assays demonstrated moderate ice recrystallisation inhibition activity, reducing relative ice crystal mean grain size to approximately 90% of the control, and significantly enhanced freeze\u2013thaw survival, with recombinant cells retaining 67% viability after three freeze\u2013thaw cycles compared with 27% for the empty-vector control. These findings identify PL002-1792 as a novel antifreeze-like protein and highlight the utility of structure-guided approaches for uncovering previously uncharacterized cold-adaptation mechanisms in polar microorganisms.<\/jats:p>","DOI":"10.3390\/microorganisms14071571","type":"journal-article","created":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T00:47:32Z","timestamp":1784508452000},"page":"1571","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Structure-Guided Discovery of a Cold-Responsive Antifreeze-like Protein from Antarctic Flavobacterium sp. PL002"],"prefix":"10.3390","volume":"14","author":[{"given":"Jennifer","family":"Charles Labo","sequence":"first","affiliation":[{"name":"Biotechnology Research Institute, Universiti Malaysia Sabah, Kota Kinabalu 88450, Sabah, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2833-274X","authenticated-orcid":false,"given":"Hui Yin","family":"Fan","sequence":"additional","affiliation":[{"name":"Food Security Research Laboratory, Faculty of Food Science and Nutrition, Universiti Malaysia Sabah, Kota Kinabalu 88400, Sabah, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hackwon","family":"Do","sequence":"additional","affiliation":[{"name":"Research Unit for Low-Temperature Convergence Technology, Korea Polar Research Institute, Incheon 21990, Republic of Korea"},{"name":"Department of Polar Sciences, University of Science and Technology, Incheon 21990, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hoang","family":"Thi Ngoc Trang","sequence":"additional","affiliation":[{"name":"Research Unit for Low-Temperature Convergence Technology, Korea Polar Research Institute, Incheon 21990, Republic of Korea"},{"name":"Department of Polar Sciences, University of Science and Technology, Incheon 21990, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8893-526X","authenticated-orcid":false,"given":"Paris","family":"Lavin","sequence":"additional","affiliation":[{"name":"Departamento de Biotecnologia, Facultad de Ciencias del Mar y Recursos Biologicos, Universidad de Antofagasta, 601 Avenida Angamos, Antofagasta 1270300, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3469-0597","authenticated-orcid":false,"given":"Mohd Faizal","family":"Abu Bakar","sequence":"additional","affiliation":[{"name":"Advanced Genomics and Bioinformatics, Malaysia Genome and Vaccine Institute, National Institutes of Biotechnology Malaysia, Jalan Bangi, Kajang 43000, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9298-7425","authenticated-orcid":false,"given":"Nur Athirah","family":"Yusof","sequence":"additional","affiliation":[{"name":"Biotechnology Research Institute, Universiti Malaysia Sabah, Kota Kinabalu 88450, Sabah, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"37519","DOI":"10.1038\/s41598-025-21587-1","article-title":"Environmental gradients shape microbial community structure and ecosystem processes in Antarctic lakes on King George Island","volume":"15","author":"Yoon","year":"2025","journal-title":"Sci. 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