Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes

Abstract Cell mass and viability are tightly linked to the productivity of fermentation processes. In 2nd generation lignocellulose-based media quantitative measurement of cell concentration is challenging because of particles, auto-fluorescence, and intrinsic colour and turbidity of the media. We s...

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Autores principales:	Ruifei Wang, Bettina Lorantfy, Salvatore Fusco, Lisbeth Olsson, Carl Johan Franzén
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1b6f5be2338e4f81a5f12b1de0a7467a
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spelling	oai:doaj.org-article:1b6f5be2338e4f81a5f12b1de0a7467a2021-12-02T15:00:19ZAnalysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes10.1038/s41598-021-90703-82045-2322https://doaj.org/article/1b6f5be2338e4f81a5f12b1de0a7467a2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90703-8https://doaj.org/toc/2045-2322Abstract Cell mass and viability are tightly linked to the productivity of fermentation processes. In 2nd generation lignocellulose-based media quantitative measurement of cell concentration is challenging because of particles, auto-fluorescence, and intrinsic colour and turbidity of the media. We systematically evaluated several methods for quantifying total and viable yeast cell concentrations to validate their use in lignocellulosic media. Several automated cell counting systems and stain-based viability tests had very limited applicability in such samples. In contrast, manual cell enumeration in a hemocytometer, plating and enumeration of colony forming units, qPCR, and in situ dielectric spectroscopy were further investigated. Parameter optimization to measurements in synthetic lignocellulosic media, which mimicked typical lignocellulosic fermentation conditions, resulted in statistically significant calibration models with good predictive capacity for these four methods. Manual enumeration of cells in a hemocytometer and of CFU were further validated for quantitative assessment of cell numbers in simultaneous saccharification and fermentation experiments on steam-exploded wheat straw. Furthermore, quantitative correlations could be established between these variables and in situ permittivity. In contrast, qPCR quantification suffered from inconsistent DNA extraction from the lignocellulosic slurries. Development of reliable and validated cell quantification methods and understanding their strengths and limitations in lignocellulosic contexts, will enable further development, optimization, and control of lignocellulose-based fermentation processes.Ruifei WangBettina LorantfySalvatore FuscoLisbeth OlssonCarl Johan FranzénNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ruifei Wang Bettina Lorantfy Salvatore Fusco Lisbeth Olsson Carl Johan Franzén Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
description	Abstract Cell mass and viability are tightly linked to the productivity of fermentation processes. In 2nd generation lignocellulose-based media quantitative measurement of cell concentration is challenging because of particles, auto-fluorescence, and intrinsic colour and turbidity of the media. We systematically evaluated several methods for quantifying total and viable yeast cell concentrations to validate their use in lignocellulosic media. Several automated cell counting systems and stain-based viability tests had very limited applicability in such samples. In contrast, manual cell enumeration in a hemocytometer, plating and enumeration of colony forming units, qPCR, and in situ dielectric spectroscopy were further investigated. Parameter optimization to measurements in synthetic lignocellulosic media, which mimicked typical lignocellulosic fermentation conditions, resulted in statistically significant calibration models with good predictive capacity for these four methods. Manual enumeration of cells in a hemocytometer and of CFU were further validated for quantitative assessment of cell numbers in simultaneous saccharification and fermentation experiments on steam-exploded wheat straw. Furthermore, quantitative correlations could be established between these variables and in situ permittivity. In contrast, qPCR quantification suffered from inconsistent DNA extraction from the lignocellulosic slurries. Development of reliable and validated cell quantification methods and understanding their strengths and limitations in lignocellulosic contexts, will enable further development, optimization, and control of lignocellulose-based fermentation processes.
format	article
author	Ruifei Wang Bettina Lorantfy Salvatore Fusco Lisbeth Olsson Carl Johan Franzén
author_facet	Ruifei Wang Bettina Lorantfy Salvatore Fusco Lisbeth Olsson Carl Johan Franzén
author_sort	Ruifei Wang
title	Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
title_short	Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
title_full	Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
title_fullStr	Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
title_full_unstemmed	Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
title_sort	analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1b6f5be2338e4f81a5f12b1de0a7467a
work_keys_str_mv	AT ruifeiwang analysisofmethodsforquantifyingyeastcellconcentrationincomplexlignocellulosicfermentationprocesses AT bettinalorantfy analysisofmethodsforquantifyingyeastcellconcentrationincomplexlignocellulosicfermentationprocesses AT salvatorefusco analysisofmethodsforquantifyingyeastcellconcentrationincomplexlignocellulosicfermentationprocesses AT lisbetholsson analysisofmethodsforquantifyingyeastcellconcentrationincomplexlignocellulosicfermentationprocesses AT carljohanfranzen analysisofmethodsforquantifyingyeastcellconcentrationincomplexlignocellulosicfermentationprocesses
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Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes

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