Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes
Artikel i vetenskaplig tidskrift, 2021

Cell mass and viability are tightly linked to the productivity of fermentation processes. In 2 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.

Författare

Ruifei Wang

Nouryon AB

Chalmers, Biologi och bioteknik, Industriell bioteknik

Bettina Lorantfy

BioPhero ApS

Chalmers, Biologi och bioteknik, Industriell bioteknik

Salvatore Fusco

Chalmers, Biologi och bioteknik, Industriell bioteknik

Universita degli Studi di Verona

Lisbeth Olsson

Chalmers, Biologi och bioteknik, Industriell bioteknik

Carl Johan Franzén

Chalmers, Biologi och bioteknik, Industriell bioteknik

Scientific Reports

2045-2322 (ISSN)

Vol. 11 1 11293

Etanolprocess med hög torrsubstanshalt och flockbildande xylosfermenterande jäst

Energimyndigheten (P37353-1), 2013-08-01 -- 2015-06-30.

Styrkeområden

Nanovetenskap och nanoteknik (SO 2010-2017, EI 2018-)

Ämneskategorier

Biomedicinsk laboratorievetenskap/teknologi

Biofysik

Annan industriell bioteknik

DOI

10.1038/s41598-021-90703-8

Mer information

Senast uppdaterat

2021-06-08