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Brewing Chemistry Analysis – Quality Assurance –

Profile of Metabolic Enzyme CO2 Determination

Material production using microorganisms such as Yeast strain S288C and single gene deletion mutants of strain Quality assurance is only one of the attributes for brand name bottles, carbon dioxide ensures that the necessary pressure is
Saccharomyces Cerevisiae has been applied to fermented food BY4742 were cultured in media containing non-labeled or C-13 products such as beer. And quality is dependent on the quality maintained. This is an important factor for guaranteeing shelf
such as beer. These microbial cell factories have been labeled glucose for metabolic enzyme analysis. Strain S288C of the starting materials, strict production processes as well as life and fresh-tasting beer. A constant concentration of carbon
metabolically engineered by regulating the expression levels of grown in heavy C-13 labeled glucose acted as an internal specialists whose keen eyes oversee the entire production. dioxide also ensures a steady taste and consistent quality of the
enzymes. To understand these metabolic pathways, it is standard for comparison to the light C-12 mutated growth Finally, millions of consumers are testing their favorite brand of beer. Beer contains 4 – 6 g/L CO2.
important to monitor the abundance of enzymes. Metabolic strains*. Information regarding glycerol biosynthesis, ethanol beer every day. Carbon dioxide in the world of Total Organic Carbon (TOC)
enzymes can be subjected to trypsin digestion, and pathway, tricarboxylic acid (TCA) cycle, polysaccharides, and An important player in the production process and a condition analyzers is referred to as Inorganic Carbon (IC). The IC method
representative tryptic peptides selected for analysis. LC-MS/MS is amino acids was determined by LC-MS/MS analysis. In one for quality is invisible: carbon dioxide (CO2), formed during the is a suitable alternative to the classical Corning reference
a highly sensitive and specific analytical technique used for example, the LCMS-8050 measured 303 peptides of 137 proteins fermenting process. After filling the beer into barrels and method for measuring CO2*.
quantitation of specific peptides in biological samples. Unlike by monitoring 2856 MRM transitions#. The library* for
ELISA, LC/MS-based MRM (Multiple Reaction Monitoring) metabolic enzymes in yeast contains 3,584 MRM transitions,
approaches do not require an antibody for each including stable isotopes. It covers all 498 trypsin digested
protein/peptide. peptides of 228 types of enzymes derived from budding yeast.

12
Light: Culture with C Glucose
6
Tryptic
Yeast A peptide
Cultivation
12
( C Glucose) cell collection extraction digestion
Heavy: Culture with C Glucose Light Inorganic Carbon (IC) measured as CO 2 in beer by a TOC analyzer*
13
6 Shimadzu TOC-L Total Organic Carbon analyzer for the analysis of CO 2
Heavy
Tryptic MRM analysis
Yeast B peptide
Cultivation
13
( C Glucose) cell collection extraction digestion 6
Corning TOC
5.5
5
Co2 [g/L] 4.5
Pdc1 Pdc6 Adh1 Adh3
4
3.5
3
Adh6 Ald4 Ald5 Ald6 beer1 alcohol-free beer2 alcohol-free beer3 beer4 unfiltrated
beer1 beer2 beer4
CO2 concentration measured by TOC and Corning reference method for various beers*

Metabolic profile of Ethanol pathway enzyme levels in wild type (black) Shimadzu LCMS-8050 for measuring metabolic enzymes in yeast
and three deletion strains (greys) of S. cerevisiae. #

* Shimadzu Brochure C146-E275: “LC/MS/MS MRM Library for Metabolic Enzymes in Yeast”
# Shimadzu Corporation Application Data Sheet: “Quantitative proteomics of metabolic enzymes in S. cerevisiae with UFMS technologies”

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