THE FERMENTATION OF LACTOSE Biology 201-Lab Group names: Addie Lupercio, Akido Sheard, and Andrea Elguezabal Introduction

Alcoholic fermentation is anaerobic respiration through which yeast oxidizes sugars producing ethanol, CO2, and ATP as by-products (Perkins, 2012). Many food items such as beer, wine, and bread are all made through the process of fermentation. Saccharomyces cerevisiae is perhaps the most common species of yeast (USEPA, 2012). Some might say it is the most important yeast because man has used it for thousands of years in baking bread and the brewing of beer and wine and ethanol production. While yeast does oxidize monomers it is unable to break apart glycosidic bonds in certain disaccharides and polysaccharides. This process can be assisted with the addition of enzymes. Today we will test the effects of yeast added to different carbohydrates. We hypothesize that more CO2 will be produced with glucose because yeast lacks the enzymes to separate the glycosidic bonds of the dissacharides lactose and sucrose. Materials and Methods A computer was used to run the Logger Pro program to measure the levels of CO2. Four 250 mL flasks were used to hold the various substrates. First, the four flasks were filled with 100mL of four different fluids. The first flask was filled with 100mL of water to serve as a negative control, flask two was filled with100mL of glucose, flask three was filled with 100mL of lactose and flask four was filled with 100mL of sucrose. We then added 10mL of suspended yeast to each of the flasks. The computer was then turned on and the Logger Pro program was pulled up; we verified that the time setting was at 3600 seconds. After we verified the program settings were correct, we attached the CO2 sensors to the Vernier Interface. We then inserted the

CO2 sensors at the top of all four flasks. At that point, the Vernier Interface program began collecting data. Results The lab classs results in Figure 1.1 show that lactose was the substrate with the least amount of CO2 production. The substrates sucrose and glucose ended up with similar rates of CO2 production. Our results show that yeast breaks down sucrose at a comparable level to glucose while lactose showed minimal or very little reaction.

Figure 1.1

Fermentation of Carbohydrates
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Figure 1.2

Fermentation of Lactose with and without Lactase

8000 7000 Rate of C02 Production 6000 5000 4000 3000 2000 1000 0 1 15 29 43 57 71 85 99 113 127 141 155 169 183 197 211 225 239 253 267 281 295 309 Seconds Elapsed Lactose Lactose/Lactase Glucose Water

Discussion Overall, the results of the experiment did not fully support our hypothesis. An enzyme in yeast assisted in the fermentation of sucrose by breaking apart the glycosidic bonds between the glucose and the fructose. Figure 1.2 shows our results when we added the necessary enzyme, lactase, to the substrate lactose in a subsequent experiment. This enzyme is necessary in order for yeast to break down lactose. There have been other experiments that have been conducted on the human intolerance of lactose where lactase supplements have been added to their diet resulting in the more efficient break down of lactose (Guzek et al., 2008). In future experiments we can conduct two identical experiments simultaneously. We believe a good follow up experiment

would be to increase the amount of yeast added to the substrates in order to see if the levels of fermentation could increase.

Works Cited Guzek M., Stojek M., Wierzbowski J., Sulkowska A., Smoczyski M. (2008) Tolerance of lowlactose milk and supplemental lactase obtained from Aspergillus oryzaein persons with lactose intolerance. Gastroenterolgia Polska. 15(5): 305-308. Perkins, D. A Laboratory Manual for Biology 1. (2012). College of Western Idaho. 65p. US Environmental Protection Agency. (2012, September 27). Saccharomyces cerevisiae Final Risk Assessment. US Environmental Protection Agency. Retrieved from: http://www.epa.gov/biotech_rule/pubs/fra/fra002.htm