{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T19:22:14Z","timestamp":1791228134724,"version":"4.1.0"},"reference-count":25,"publisher":"Wiley","license":[{"start":{"date-parts":[[2016,1,1]],"date-time":"2016-01-01T00:00:00Z","timestamp":1451606400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NSTIP Strategic Technologies Program in the Kingdom of Saudi Arabia","award":["10-BIO1257-03"],"award-info":[{"award-number":["10-BIO1257-03"]}]},{"DOI":"10.13039\/501100004054","name":"King Abdulaziz University of Saudi Arabia","doi-asserted-by":"publisher","award":["10-BIO1257-03"],"award-info":[{"award-number":["10-BIO1257-03"]}],"id":[{"id":"10.13039\/501100004054","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["BioMed Research International"],"published-print":{"date-parts":[[2016]]},"abstract":"<jats:p>Increase in the incidence of Insulin Dependent Diabetes Mellitus (IDDM) among people from developed and developing countries has created a large global market for insulin. Moreover, exploration of new methods for insulin delivery including oral or inhalation route which require very high doses would further increase the demand of cost-effective recombinant insulin. Various bacterial and yeast strains have been optimized to overproduce important biopharmaceuticals. One of the approaches we have taken is the production of recombinant human insulin along with C-peptide in yeast<jats:italic>Pichia pastoris<\/jats:italic>. We procured a cDNA clone of insulin from Origene Inc., USA. Insulin cDNA was PCR amplified and cloned into yeast vector pPICZ-<jats:italic>\u03b1<\/jats:italic>. Cloned insulin cDNA was confirmed by restriction analysis and DNA sequencing. pPICZ-<jats:italic>\u03b1<\/jats:italic>-insulin clone was transformed into<jats:italic>Pichia pastoris SuperMan<\/jats:italic><jats:sub><jats:italic>5<\/jats:italic><\/jats:sub>strain. Several Zeocin resistant clones were obtained and integration of insulin cDNA in<jats:italic>Pichia<\/jats:italic>genome was confirmed by PCR using insulin specific primers. Expression of insulin in<jats:italic>Pichia<\/jats:italic>clones was confirmed by ELISA, SDS-PAGE, and Western blot analysis. In vivo efficacy studies in streptozotocin induced diabetic mice confirmed the activity of recombinant insulin. In conclusion, a biologically active human proinsulin along with C-peptide was expressed at high level using<jats:italic>Pichia pastoris<\/jats:italic>expression system.<\/jats:p>","DOI":"10.1155\/2016\/3423685","type":"journal-article","created":{"date-parts":[[2016,8,4]],"date-time":"2016-08-04T21:04:54Z","timestamp":1470344694000},"page":"1-7","source":"Crossref","is-referenced-by-count":25,"title":["Expression and Purification of C-Peptide Containing Insulin Using<i>Pichia pastoris<\/i>Expression System"],"prefix":"10.1155","volume":"2016","author":[{"given":"Mohammed N.","family":"Baeshen","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"},{"name":"Department of Biological Sciences, Faculty of Science, University of Jeddah, P.O. Box 80327, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thamer A. F.","family":"Bouback","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mubarak A.","family":"Alzubaidi","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5586-3829","authenticated-orcid":true,"given":"Roop S.","family":"Bora","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"},{"name":"Department of Biotechnology, Eternal University, Baru Sahib, Himachal Pradesh 173 101, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3729-0374","authenticated-orcid":true,"given":"Mohammed A. T.","family":"Alotaibi","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Omar T. O.","family":"Alabbas","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sultan M.","family":"Alshahrani","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed A. M.","family":"Aljohani","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rayan A. A.","family":"Munshi","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed","family":"Al-Hejin","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8316-9714","authenticated-orcid":true,"given":"Mohamed M. M.","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"},{"name":"Nucleic Acids Research Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City of Scientific Research and Technology Applications, Alexandria 21934, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elrashdy M.","family":"Redwan","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"},{"name":"Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technology Applications, Alexandria 21934, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hassan A. I.","family":"Ramadan","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"},{"name":"Cell Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Tahrir Street, Dokki, Cairo 12311, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kulvinder S.","family":"Saini","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"},{"name":"Department of Biotechnology, Eternal University, Baru Sahib, Himachal Pradesh 173 101, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nabih A.","family":"Baeshen","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","reference":[{"issue":"5","key":"18","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1002\/j.1875-9114.1998.tb03927.x","volume":"18","year":"1998","journal-title":"Pharmacotherapy"},{"key":"26","doi-asserted-by":"publisher","DOI":"10.2337\/diacare.27.5.1047"},{"key":"21","doi-asserted-by":"publisher","DOI":"10.1210\/er.22.5.706"},{"key":"24","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-004-1809-x"},{"issue":"3","key":"5","first-page":"133","volume":"16","year":"1993","journal-title":"Diabetes Care"},{"key":"1","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-014-0141-0"},{"key":"6","first-page":"149","volume-title":"Insulin lispro (Humalog)","year":"1999"},{"key":"2","doi-asserted-by":"publisher","DOI":"10.4014\/jmb.1412.12079"},{"key":"16","first-page":"265","volume":"9","year":"2006","journal-title":"Arab Journal of Biotechnology"},{"key":"17","doi-asserted-by":"publisher","DOI":"10.1080\/10826060701774312"},{"key":"8","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.5.5.1111"},{"key":"19","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1287-1305"},{"key":"9","doi-asserted-by":"publisher","DOI":"10.1002\/yea.320050206"},{"key":"20","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/15.9.3859"},{"key":"7","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0893-905"},{"key":"3","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2010.00582.x"},{"key":"14","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2005.00159.x"},{"key":"10","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2859-9-31"},{"issue":"1","key":"11","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1111\/j.1470-8744.1999.tb01151.x","volume":"29","year":"1999","journal-title":"Biotechnology and Applied Biochemistry"},{"key":"12","doi-asserted-by":"publisher","DOI":"10.1007\/s002530000402"},{"key":"13","doi-asserted-by":"publisher","DOI":"10.1007\/s10529-005-1007-7"},{"issue":"1","key":"25","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1002\/1097-0290(20010405)73:1<74::AID-BIT1038>3.0.CO;2-V","volume":"73","year":"2001","journal-title":"Biotechnology and Bioengineering"},{"key":"22","doi-asserted-by":"publisher","DOI":"10.1111\/j.1475-097x.2004.00558.x"},{"key":"15","doi-asserted-by":"publisher","DOI":"10.1155\/2008\/281536"},{"key":"23","doi-asserted-by":"publisher","DOI":"10.1007\/s00125-006-0559-y"}],"container-title":["BioMed Research International"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/downloads.hindawi.com\/journals\/bmri\/2016\/3423685.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/downloads.hindawi.com\/journals\/bmri\/2016\/3423685.xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/downloads.hindawi.com\/journals\/bmri\/2016\/3423685.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,18]],"date-time":"2024-06-18T18:05:54Z","timestamp":1718733954000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.hindawi.com\/journals\/bmri\/2016\/3423685\/"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016]]},"references-count":25,"alternative-id":["3423685","3423685"],"URL":"https:\/\/doi.org\/10.1155\/2016\/3423685","relation":{},"ISSN":["2314-6133","2314-6141"],"issn-type":[{"value":"2314-6133","type":"print"},{"value":"2314-6141","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016]]}}}