{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T08:51:30Z","timestamp":1791363090734,"version":"4.3.1"},"reference-count":58,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T00:00:00Z","timestamp":1787961600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"vor","delay-in-days":39,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Appl Microbiol Biotechnol"],"abstract":"<jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>\n                      Fibrillar type III collagen, characterized by a distinct triple helix structure, is a critical structural component of the mammalian extracellular matrix (ECM). Its unique mechanical properties and biological functions make it a highly valuable biomaterial for medical applications. However, the limited availability of natural human type III collagen (hCOL3) and the defects of animal-derived sources necessitate sustainable alternatives. Here, we developed an integrated molecular engineering strategy for the efficient production of full-length recombinant human type III collagen (rhCOL3) in\n                      <jats:italic>Komagataella phaffii<\/jats:italic>\n                      . The N- and C-terminal propeptides of the rhCOL3\u03b11 chain were rationally redesigned to preserve its intrinsic capability for triple helix formation. Synergistic co-expression of human P4H\u03b1\u03b2 and\n                      <jats:italic>Bacillus anthracis<\/jats:italic>\n                      -derived BaP4H achieved a proline hydroxylation level of 52.88%, enabling the stable assembly of rhCOL3 procollagen homotrimers (&gt;\u2009300\u00a0kDa). Subsequent optimization, including high-copy-number strain screening, signal sequence substitution (with Ost1 identified as optimal), and co-expression of the molecular chaperones (e.g., BCY1 and ERO1), significantly enhanced productivity. The engineered strain achieved a rhCOL3 procollagen yield of 430\u00a0mg\/L in a 5-L fed-batch bioreactor. The purified rhCOL3 exhibited a stable triple helix conformation, a melting temperature of 38.87\u00a0\u00b0C, and the ability to self-assemble into fibrils with a characteristic D-banding pattern. Biological assays confirmed that the rhCOL3 promotes cell proliferation, adhesion, and migration at levels comparable to those of natural collagen. This study establishes a promising platform for the industrial-scale production of bioactive, triple-helical human collagen.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Key points<\/jats:title>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>Synergistic P4H\u03b1\u03b2 and BaP4H co-expression achieves 52.88% Hyp level and Tm 38.87\u00a0\u00b0C.<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>Ost1 and Bcy1 co-expression elevated rhCOL3 yield to 430\u00a0mg\/L in a 5-L bioreactor.<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>Purified rhCOL3 self-assembles into D-banded fibrils resembling native collagen.<\/jats:italic>\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s00253-026-13996-7","type":"journal-article","created":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T22:44:11Z","timestamp":1787957051000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Enhanced biosynthesis of full-length triple-helical human type III collagen in engineered Komagataella phaffii"],"prefix":"10.1007","volume":"110","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7626-3224","authenticated-orcid":false,"given":"Zhijian","family":"Ni","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaolu","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Da","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoxi","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"ZhiDa","family":"Wei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulei","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yao","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunting","family":"Ye","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunlin","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,29]]},"reference":[{"key":"13996_CR1","doi-asserted-by":"publisher","first-page":"413","DOI":"10.1002\/biot.200900233","volume":"5","author":"S Abad","year":"2010","unstructured":"Abad S, Kitz K, H\u00f6rmann A, Schreiner U, Hartner FS, Glieder A (2010) Real-time PCR-based determination of gene copy numbers in Pichia pastoris. 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