Development of chemically defined media to express trp-analog-labeled proteins in a lactococcus lactis trp auxotroph

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dc.contributor.authorShao, Jinfeng
dc.contributor.authorMarcondes, Marcelo F. M. [UNIFESP]
dc.contributor.authorOliveira, Vitor [UNIFESP]
dc.contributor.authorBroos, Jaap
dc.date.accessioned2019-01-21T10:29:52Z
dc.date.available2019-01-21T10:29:52Z
dc.date.issued2016
dc.description.abstractChemically defined media for growth of Lactococcus lactis strains contain about 50 components, making them laborious and expensive growth media. However, they are crucial for metabolism studies as well as for expression of heterologous proteins labeled with unnatural amino acids. In particular, the L. lactis Trp auxotroph PA1002, over-expressing the tryptophanyl tRNA synthetase enzyme of L. lactis, is very suitable for the biosynthetic incorporation of Trp analogs in proteins because of its most relaxed substrate specificity reported towards Trp analogs. Here we present two much simpler defined media for L. lactis, which consist of only 24 or 31 components, respectively, and with which the L. lactis Trp auxotroph shows similar growth characteristics as with a 50-component chemically defined medium. Importantly, the expression levels of two recombinant proteins used for evaluation were up to 2-3 times higher in these new media than in the 50-component medium, without affecting the Trp analog incorporation efficiency. Taken together, the simplest chemically defined media reported so far for L. lactis are presented. Since L. lactis also shows auxotrophy for Arg, His, Ile, Leu Val, and Met, our simplified media may also be useful for the biosynthetic incorporation of analogs of these five amino acids. (C) 2016 The Author(s) Published by S. Karger AG, Baselen
dc.description.affiliationLaboratory of Biophysical Chemistry and Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Groningen, The Netherlands
dc.description.affiliationDepartment of Biophysics, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil
dc.description.affiliationUnifespDepartment of Biophysics, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipChina Scholarship Council
dc.description.sponsorshipFAPESP
dc.description.sponsorshipIDThis work was financially supported by the China Scholarship Council and FAPESP. We thank Dr. B.W. Dijkstra for critical reading of the manuscript.
dc.format.extent269-276
dc.identifierhttps://doi.org/10.1159/000445687
dc.identifier.citationJournal Of Molecular Microbiology And Biotechnology. Basel, v. 26, n. 4, p. 269-276, 2016.
dc.identifier.doi10.1159/000445687
dc.identifier.issn1464-1801
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/49447
dc.identifier.wosWOS:000381109500004
dc.language.isoeng
dc.publisherPontificia Universidade Catolica Campinas
dc.relation.ispartofJournal Of Molecular Microbiology And Biotechnology
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectAlloproteinen
dc.subjectLactococcus Lactisen
dc.subjectSynthetic Mediumen
dc.subjectTryptophan AnalogTryptophan Analogsen
dc.subjectBiosynthetic Incorporationen
dc.subjectFluorescence Spectroscopyen
dc.subjectStreptococcus-Cremorisen
dc.subjectGrowthen
dc.subjectOverproductionen
dc.subjectRequirementsen
dc.subjectHosten
dc.titleDevelopment of chemically defined media to express trp-analog-labeled proteins in a lactococcus lactis trp auxotrophen
dc.typeinfo:eu-repo/semantics/article
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