{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,5]],"date-time":"2026-04-05T17:00:50Z","timestamp":1775408450511,"version":"3.50.1"},"reference-count":49,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2018,3,26]],"date-time":"2018-03-26T00:00:00Z","timestamp":1522022400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["ERA\u2010IB\/0001\/2012"],"award-info":[{"award-number":["ERA\u2010IB\/0001\/2012"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["INST 20876\/127\u20101 FUGG"],"award-info":[{"award-number":["INST 20876\/127\u20101 FUGG"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2018,7]]},"abstract":"<jats:sec><jats:label\/><jats:p><jats:italic>Medicago truncatula<\/jats:italic> is an established model for studying legume biology. More recently, it has also been exploited as a Molecular Farming platform for the production of recombinant proteins, with the successful expression of fungal and human proteins in plants and cell suspension cultures of this species. One of the challenges that now must be overcome is the degradation of final products during production and downstream processing stages. In the <jats:italic>M. truncatula<\/jats:italic> genome, there are more than 400 putative protease\u2010encoding genes, but to date, the proteolytic content of Medicago cell cultures has not been studied. In this report, the proteolytic activities that can potentially hamper the successful production of recombinant proteins in this system are evaluated. The potential proteases responsible for the degradation of target proteins are identified. Interestingly, the number of proteases found in Medicago spent medium is considerably lower than that of the well\u2010established tobacco bright yellow 2 (BY\u20102) system. Papain\u2010like cysteine proteases are found to be the major contributors to recombinant protein degradation in Medicago. This knowledge is used to engineer a cell line with reduced endogenous protease activity by expressing a selective protease inhibitor, further improving this expression platform.<\/jats:p><\/jats:sec>","DOI":"10.1002\/biot.201800050","type":"journal-article","created":{"date-parts":[[2018,3,12]],"date-time":"2018-03-12T07:22:39Z","timestamp":1520839359000},"source":"Crossref","is-referenced-by-count":18,"title":["Low Protease Content in <i>Medicago truncatula<\/i> Cell Cultures Facilitates Recombinant Protein Production"],"prefix":"10.1002","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7809-6813","authenticated-orcid":false,"given":"Rita B.","family":"Santos","sequence":"first","affiliation":[{"name":"Plant Cell Biology Laboratory, Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa Av. da Rep\u00fablica 2780\u2010157 Oeiras Portugal"}]},{"given":"Balakumaran","family":"Chandrasekar","sequence":"additional","affiliation":[{"name":"The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford South Parks Road Oxford OX1 3RB United Kingdom"}]},{"given":"Manoj K.","family":"Mandal","sequence":"additional","affiliation":[{"name":"Department of Plant Biotechnology, Fraunhofer Institute for Molecular Biology and Applied Ecology IME Forckenbeckstrasse 6 52074 Aachen Germany"}]},{"given":"Farnusch","family":"Kaschani","sequence":"additional","affiliation":[{"name":"Chemical Biology, Faculty of Biology, University of Duisburg\u2010Essen, ZMB Universit\u00e4tsstra\u00dfe 2 45117 Essen Germany"}]},{"given":"Markus","family":"Kaiser","sequence":"additional","affiliation":[{"name":"Chemical Biology, Faculty of Biology, University of Duisburg\u2010Essen, ZMB Universit\u00e4tsstra\u00dfe 2 45117 Essen Germany"}]},{"given":"Leonard","family":"Both","sequence":"additional","affiliation":[{"name":"The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford South Parks Road Oxford OX1 3RB United Kingdom"}]},{"given":"Renier A. 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