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In situ near infrared spectroscopy monitoring of cyprosin production by recombinant Saccharomyces cerevisiae strains

2014-10-20Journal article Restricted access
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dc.contributor.authorSampaio, Pedro
dc.contributor.authorSales, Kevin C.
dc.contributor.authorRosa, Filipa O.
dc.contributor.authorLopes, Marta B.
dc.contributor.authorCalado, Cecília
dc.date.accessioned2020-07-31T16:57:42Z
dc.date.available2020-07-31T16:57:42Z
dc.date.issued2014-10-20
dc.description.abstractNear infrared (NIR) spectroscopy was used to in situ monitoring the cultivation of two recombinant Saccharomyces cerevisiae strains producing heterologous cyprosin B. NIR spectroscopy is a fast and non-destructive technique, that by being based on overtones and combinations of molecular vibrations requires chemometrics tools, such as partial least squares (PLS) regression models, to extract quantitative information concerning the variables of interest from the spectral data. In the present work, good PLS calibration models based on specific regions of the NIR spectral data were built for estimating the critical variables of the cyprosin production process: biomass concentration, cyprosin activity, cyprosin specific activity, the carbon sources glucose and galactose concentration and the by-products acetic acid and ethanol concentration. The PLS models developed are valid for both recombinant S. cerevisiae strains, presenting distinct cyprosin production capacities, and therefore can be used, not only for the real-time control of both processes, but also in optimization protocols. The PLS model for biomass yielded a R2 = 0.98 and a RMSEP = 0.46 g dcw l−1, representing an error of 4% for a calibration range between 0.44 and 13.75 g dcw l−1. A R2 = 0.94 and a RMSEP = 167 U ml−1 were obtained for the cyprosin activity, corresponding to an error of 6.7% of the experimental data range (0–2509 U ml−1), whereas a R2 = 0.93 and RMSEP = 672 U mg−1 were obtained for the cyprosin specific activity, corresponding to an error of 7% of the experimental data range (0–11,690 U mg−1). For the carbon sources glucose and galactose, a R2 = 0.96 and a RMSECV of 1.26 and 0.55 g l−1, respectively, were obtained, showing high predictive capabilities within the range of 0–20 g l−1. For the metabolites resulting from the cell growth, the PLS model for acetate was characterized by a R2 = 0.92 and a RMSEP = 0.06 g l−1, which corresponds to a 6.1% error within the range of 0.41–1.23 g l−1; for the ethanol, a high accuracy PLS model with a R2 = 0.97 and a RMSEP = 1.08 g l−1 was obtained, representing an error of 9% within the range of 0.18–21.76 g l−1. The present study shows that it is possible the in situ monitoring and prediction of the critical variables of the recombinant cyprosin B production process by NIR spectroscopy, which can be applied in process control in real-time and in optimization protocols. From the above, NIR spectroscopy appears as a valuable analytical tool for online monitoring of cultivation processes, in a fast, accurate and reproducible operation mode.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationSAMPAIO, Pedro N.; [et al] – In situ near infrared spectroscopy monitoring of cyprosin production by recombinant Saccharomyces cerevisiae strains. Journal of Biotechnology. ISSN 0168-1656. Vol. 188 (2014), pp. 148-157pt_PT
dc.identifier.doi10.1016/j.jbiotec.2014.07.454pt_PT
dc.identifier.issn0168-1656
dc.identifier.urihttp://hdl.handle.net/10400.21/12133
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://pdf.sciencedirectassets.com/271109/1-s2.0-S0168165614X00179/1-s2.0-S0168165614008037/main.pdf?X-Amz-Date=20200731T162446Z&X-Amz-Algorithm=AWS4-HMAC-SHA256&X-Amz-Signature=93fc6d56213fce46465330d11891beeea61e3f76b734dc66f02887f6a63b0b40&X-Amz-Credential=ASIAQ3PHCVTYXI6TQ2VB%2F20200731%2Fus-east-1%2Fs3%2Faws4_request&type=client&tid=prr-72e3ce82-f4bd-400e-af28-b109e2895356&sid=ba0a29565fa23547ca9be220ab8265222faagxrqb&pii=S0168165614008037&X-Amz-SignedHeaders=host&X-Amz-Security-Token=IQoJb3JpZ2luX2VjEID%2F%2F%2F%2F%2F%2F%2F%2F%2F%2FwEaCXVzLWVhc3QtMSJGMEQCIHmqUpjR4vNeGZkYUVxOOZogM91on954YQ93R%2BsgbcpQAiBSmdOaYqGibkxkmSXTx4HtCvW5Tix7LmFu4MKoK7Sjniq0AwhJEAMaDDA1OTAwMzU0Njg2NSIMQ2xRcGbGU60M1Oj6KpEDvHLf9jCw5RHVVPw5l51Le1HANaHuMc%2Fx%2FHj%2FRHrs0t6GPHPEZxcpqcct7jAvH7Efxe%2BrmZRyQpx1iaEeypPSXwxdya76rPCz%2B9yRI5Q4dNc8WSjvWz9Wx1yVBxy02O41UPXuCXVi6sg4gSr2sLCXkFa%2BvLbehBF0p%2BbLRT%2FW1wUJAXFePSHzlyKUMC5Sva5amO7Kx7ZG8jL0XRSOPC4l03QZMqatVJHeheSk34vmG1aASXSur4gaoYAbffZDg6LBV2sEVYqhivPC%2BoZST80jewOtbbmMcPiZsMRwJf0MWmr4waXRzmrEFie9vOvAFbvDRX26qIwQSZQH9mFnj02pD%2FG4rHGX85lM0jyxVQ12fqdcc5lQIYdBuxvpBYjV3kcVFpMmYEZ8guyPns4Svb%2B4XEQflHKdtsmhqAaXuHByGx9OFFSzVe4doMzuxt480UFCMNeLysezMOPXvlhr6GeThmbz4LRHD%2FPERUlZkuJWecuVSXH9RDWS6LwXrhEz%2BL%2FBtDdnXD6zyEGxyen7tP%2FwaAswmfWQ%2BQU67AEatnlkMO5bK1ylC902CggLa8Rs2RkdnXZfbmsYlnA%2FM8thQDCz2cp9EH4h2IEGZv7lTg5FLLaQbopZCQ0hQXVr3y2vZPGm1O2AEIZAt%2FyVfDkg3wu6veydRwrbJABFm0sj4WFCtCQ9TmziINTSd8UscIYrZ4oZ49Uz6O7rW7dW72VoJGSib0s%2BwgFd0HFy6SY3ic6vkBEK9eJGTQ5tGBrAh90DlYIOORJwJCX23Kqm4oKBoTLJazQ1aTspJ1maRx4aJly1FrH%2FdRptExn%2BUZPzpsNSNPZc%2FdrR0184Cn7Ibcu6hm%2FFITNd%2BmiQxQ%3D%3D&host=68042c943591013ac2b2430a89b270f6af2c76d8dfd086a07176afe7c76c2c61&X-Amz-Expires=300&hash=ff86ec15850fd56ffcface33e2dd0131421a0e91e6265b24597553a629506284pt_PT
dc.subjectIn situ monitoringpt_PT
dc.subjectNear infraredpt_PT
dc.subjectPartial least squares regressionpt_PT
dc.subjectRecombinant cyprosin Bpt_PT
dc.subjectSaccharomyces cerevisiaept_PT
dc.titleIn situ near infrared spectroscopy monitoring of cyprosin production by recombinant Saccharomyces cerevisiae strainspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F73758%2F2010/PT
oaire.citation.endPage157pt_PT
oaire.citation.startPage148pt_PT
oaire.citation.titleJournal of Biotechnologypt_PT
oaire.citation.volume188pt_PT
oaire.fundingStreamSFRH
person.familyNameSampaio
person.familyNameB. Lopes
person.familyNameCalado
person.givenNamePedro
person.givenNameMarta
person.givenNameCecília
person.identifier1251979
person.identifier1195979
person.identifier130332
person.identifier.ciencia-idAF12-6ABA-43D8
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person.identifier.ciencia-id9418-E320-3177
person.identifier.orcid0000-0003-2917-4904
person.identifier.orcid0000-0002-4135-1857
person.identifier.orcid0000-0002-5264-9755
person.identifier.ridF-5378-2011
person.identifier.ridE-2102-2014
person.identifier.scopus-author-id24178064100
person.identifier.scopus-author-id55489480400
person.identifier.scopus-author-id6603163260
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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