High-gravity brewing is a commonly used technique in beer production duo to its high economic efficiency.During high-gravity brewing, brewer’s yeast is subjected to different environmental stresses in different brewing stages.Proline is considered as an important stress-protective agent in brewer’s yeast, which endows brewer’s yeast with better tolerance ability against stresses in high-gravity brewing.Since the intracellular concentration of proline can be used as a reference for screening of brewing yeast with good environmental tolerance ability, the establishment of a method capable of rapidly detecting the concentration of intracellular proline in brewing yeast is required.This study developed a novel fluorescent microplate method (FMM) for the detection of intracellular proline concentration in brewing yeasts based on sodium hypochlorite oxidation and o-phthalaldehyde (OPA)/n-acetyl-l-cysteine (NAC) fluorescence methods combined with the optimization of cells disruption technique.The linear ranges and the recovery rate of the novel method were 5-40 μmol/L (0.58-4.6 mg/L) and 73.33%-105.5%, respectively.Compared to the high-performance liquid chromatography method, the FMM assay had no significantly different absolute quantification of intracellular proline in brewing yeasts, but the procedures in the FMM assay were much faster and more convenient.Therefore, the novel FMM method proposed in this study not only proposes a new solution for the detection of intracellular proline in brewing yeasts, but also provides a new strategy for screening the proline-accumulating brewer’s yeasts with better adaptation to stresses in high-gravity brewing.
YU Yingying
,
XU Ruiqi
,
NIU Chengtuo
,
WANG Jinjing
,
ZHENG Feiyun
,
LIU Chunfeng
,
LI Qi
. Development of a novel detection method of intracellular proline in brewer’s yeast based on fluorescent microtiter plate[J]. Food and Fermentation Industries, 2024
, 50(20)
: 321
-325
.
DOI: 10.13995/j.cnki.11-1802/ts.038638
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