{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T19:37:44Z","timestamp":1790019464664,"version":"4.0.1"},"reference-count":27,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2005,10,27]],"date-time":"2005-10-27T00:00:00Z","timestamp":1130371200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp;amp; Bioengineering"],"published-print":{"date-parts":[[2006,3,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>A potential vaccine candidate, <jats:italic>Necator americanus<\/jats:italic> secretory protein (Na\u2010ASP1), against hookworm infections, has been expressed in <jats:italic>Pichia<\/jats:italic> <jats:italic>pastoris.<\/jats:italic> Na\u2010ASP1, a 45 kDa protein containing 20 cysteines, was directed outside the cell by fusing the protein to the preprosequence of the \u03b1\u2010mating factor of <jats:italic>Saccharomyces cerevisiae<\/jats:italic>. Most of the protein produced by single copy clones was secreted outside the cell. However, increasing gene copy number of Na\u2010ASP1 protein in <jats:italic>P. pastoris<\/jats:italic> saturated secretory capacity and therefore, decreased the amount of secreted protein in clones harboring multiple copies of Na\u2010ASP1 gene. Overexpression of the endoplasmic reticulum (ER) resident, homologous chaperone protein, protein disulfide isomerase (PDI) was able to increase the secretion of (Na\u2010ASP1) protein in high copy clones. The effect of PDI levels on secretion of Na\u2010ASP1 protein was examined in clones with varying copy number of PDI gene. Increase in secreted Na\u2010ASP1 secretion is correlated well with the PDI copy number. Increasing levels of PDI also increased overall Na\u2010ASP1 protein production in all the clones. Nevertheless, there was still accumulation of intracellular Na\u2010ASP1 protein in <jats:italic>P. pastoris<\/jats:italic> clones over\u2010expressing Na\u2010ASP1 and PDI proteins. \u00a9 2005 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/bit.20762","type":"journal-article","created":{"date-parts":[[2005,10,27]],"date-time":"2005-10-27T20:50:00Z","timestamp":1130446200000},"page":"771-778","source":"Crossref","is-referenced-by-count":159,"title":["Enhancement of protein secretion in <i>Pichia pastoris<\/i> by overexpression of protein disulfide isomerase"],"prefix":"10.1002","volume":"93","author":[{"given":"Mehmet","family":"Inan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dinesh","family":"Aryasomayajula","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jayanta","family":"Sinha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael M.","family":"Meagher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2005,10,27]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1749-6632.1990.tb24257.x"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.10771"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0958-1669(02)00330-0"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(91)90152-2"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1139\/b95-336"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(84)90077-9"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmr.687"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp000107q"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm1052"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/35017001"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/355033a0"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(98)90005-2"},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/0962-8924(92)90309-B"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(96)00028-2"},{"key":"e_1_2_1_16_1","volume-title":"Pichia protocols","author":"Inan M","year":"2005"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1006\/prep.1995.1092"},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1006\/scdb.2000.0160"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/yea.1208"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.66.2.775-782.2000"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1006\/prep.1995.1071"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0494-381"},{"key":"e_1_2_1_23_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0061(19970330)13:4<337::AID-YEA98>3.0.CO;2-K"},{"key":"e_1_2_1_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.10367"},{"key":"e_1_2_1_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0079-6107(03)00019-1"},{"key":"e_1_2_1_26_1","doi-asserted-by":"publisher","DOI":"10.1006\/prep.2000.1335"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.2001.4479"},{"key":"e_1_2_1_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02942184"}],"container-title":["Biotechnology and Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fbit.20762","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.20762","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:34:28Z","timestamp":1760056468000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/bit.20762"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,10,27]]},"references-count":27,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2006,3,5]]}},"alternative-id":["10.1002\/bit.20762"],"URL":"https:\/\/doi.org\/10.1002\/bit.20762","archive":["Portico"],"relation":{},"ISSN":["0006-3592","1097-0290"],"issn-type":[{"value":"0006-3592","type":"print"},{"value":"1097-0290","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,10,27]]}}}