Measurement of Glucose Content of Yeast Diet Components & Monitoring of Glucose Consumption in the Yeast Growth Projects


Class: BE210
Group: W9
Members:

Noura Selim
Andrea Levine
Shirley Duarte
Edward Yang
Ranliang Hu

Date: April 2004

Full Text

Abstract: 

The growth of Saccharomyces cerevisiae, commonly referred to as Baker’s Yeast, depends on both the presence of as well as the type of nutrients. In the lag phase of growth, the yeast adjusts to their new environment by synthesizing new enzymes and proteins. Nutrients are consumed during the lag phase. Depending on the type of nutrients present, the length of this phase will vary. If the yeast is grown long enough, it will experience a few lag phases depending on the number of different nutrients present. In the log phase, the yeast also consumes nutrients, which are present in excess, as they are needed for replication. However, as the cycle continues, nutrient depletion occurs. S. cervisiae is dependent on glucose as a primary form of energy. In order to provide the yeast with nutrients, the typical yeast diet consists of dextrose, peptone, and yeast extract. These different diet components represent different forms of nutrients. Dextrose is a form of glucose. Peptone is derived from proteins, and yeast extract is produced from autolysed yeast cells. In order to measure the yeast’s glucose consumption, a glucose (HK) assay kit will be used. This kit measures glucose consumption through the change in absorbance that occurs due to the phosphorylation of glucose and the reduction of NADH. Specifically, first glucose is phosphorylated using ATP. The inorganic phosphate binds to the glucose, making it glucose-6-phosphate. This reaction is catalyzed by hexokinase. In the final reaction, glucose-6-phosphate is reacted with NAD to form 6-phosphogluconate and NADH. This reaction is catalyzed by glucose-6-phosphate dehydrogenase. The reduction of NAD to NADH causes an increase in absorbance at 340 nanometers, which is proportional to the change in glucose concentration. The change in glucose concentration over time would equal the yeast’s consumption of glucose.