B Fructofuranosidase Lab Report

gluconeogenesis 21. sucrose Part II: Putting It All Together Multiple Choice 1. A 2.

• Mutations in the ALDOB gene reduce the function of the enzyme, impairing the ability to metabolize fructose.

During this experiment, sugar sources were varied and respiration rate evaluated. To begin, a water bath was set at 30 degrees Celsius. This creates an optimum temperature for the enzymes in yeast to breakdown sugar and give off CO₂. Each sugar source, glucose, sucrose, lactose and glycerol were all added to its own unique yeast sample, one at a time. Each sugar source that was added to the yeast solution was immediately incubated for 10 min, then was transferred to a respiration chamber. The CO₂ sensor was put in, recording the CO₂ respiration for 4 min. This process was done for each sugar source. The reparation rate was recorded through Logger Pro. After 4 min passed, the slope was recorded, resulting in respiration rate.

Lactose is a sugar that can be put into smaller molecules, glucose and galactose. Lactose is when you are not able to digest milk and dairy meaning that the enzyme lactase that breaks down lactose is not functioning properly. ONPG was used as a substitute for lactase because even though it is colorless it helps show enzyme activity by turning yellow. This experiment measured the absorbance ONPG when exposed to lactase within an environment of different salinity’s. The enzyme, lactase, was obtained by crushing a lactaid pill and then was added into four cuvettes. ONPG and salt solution of different concentrations were added and their levels of absorption was measured by a spectrophotometer. The results showed that higher salt concentrations have a lower level of absorption. There were 4 cuvettes and within those cuvettes that solutions within them were being tested and the results showed the more salt solution added with the lactase the lower the absorbance. The less salt solution there was a higher rate of absorbance. The data supported the hypothesis that with increasing NaCl concentration there would be a decrease in enzyme activity.

Enzymes react differently under different conditions and concentrations, being the most productive at the enzymes specific optimum condition and concentration. The enzyme sucrase, extracted from yeast, breaks down the complex sugar sucrose into the simple sugar glucose. Testing for sucrase’s optimum environment, multiple reactions were ran using varying amounts and concentrations of sucrose and sucrase at different pHs and temperatures. The product was then treated with Benedicts solution to visually observe what amount of glucose was present after the reaction was ran; negative results being little to no glucose present and positive results being

The hypothesis stats that as the sucrose concentration is increased, rate of respiration will increase and therefore the CO2 production of yeast cells will rise. Sucrose is a disaccharide composed of the monosaccharaides glucose and fructose. Glucose is a reactant in anaerobic cell respiration. In the absence of oxygen, glucose will react with the yeast producing ethanol and CO2.

reaction rate increases. If the temperature of an enzyme gets to high the reaction rate will slow

40 mL of a concentrated solution of sucrose was prepared at 200mg/mL. Using appropriate dilutions of the stock, 11 solutions including a control solution were made in plastic tubes. The enzyme reaction with sucrose was run in 2 mL volume at room temperature in water. The enzyme constituted half the volume of the stock solution. The substrate was added to the enzyme in order to start the reaction. Each reaction ran for 5 min after which 2 mLs of DNS reagent was added. The solution was boiled for 10 min and the results were read using a spectrophotometer.

Chemical kinetics involving reaction rates and mechanisms is an essential part of our daily life in the modern world. It helps us understand whether particular reactions are favorable and how to save time or prolong time during each reaction. Experiment demonstrated the how concentration, temperature and presence of a catalyst can change the rate of a reaction. 5 runs of dilution and reaction were made to show the effect of concentration on chemical reactions. A certain run from the previous task was twice duplicated to for a “hot and cold” test for reaction rate. The prior run was again duplicated for a test with

Enzyme catalysis is dependant upon factors such as concentration of enzyme and substrate, temperature and pH. These factors determine the rate of reaction, and an increase in temperature or pH above the optimum will

Invertase, also known as beta-fructofuranosidase is a digestive enzyme that catalyses the hydrolysis (breakdown) of sucrose (table sugar) to glucose.. It is used in combination with other carbohydrates to enhance overall starch and sugar digestion. Invertase can hydrolyse the connection between glucose and fructose; this means that it uses water break sucrose into to separate monomers. This leaders the enzyme to be inhibited by ‘increased concentrations of sucrose of invertase.’

The hypothesis is as the substrate concentration has an increase so will the reaction of velocity if the amount of enzyme is kept constant.

Typically, “The yeasts in the mixture metabolize sucrose into glucose and fructose, then into ethanol and carbon dioxide. Ethanol is then oxidized by the bacteria (in the presence of air) to acetaldehyde, then to acetic acid.” (Nummer, 2016) Furthermore, “The acetic acid bacteria also utilize glucose to produce gluconic acid to approximately 2%. Fructose is used to a lesser extent and some remains after the fermentation. Some glucose will remain unmetabolized, and together with the remaining fructose, provides sweetness.” (Nummer, 2016) Also, The pH of the kombucha is reduced as it brews, first starting around 5, then, Nummer's study shows, “Once fermentation starts that pH is reduced in approximately seven days to a finishing pH of [less than or equal to] 2.5.” (Nummer, 2016)

Enzymes are natural catalysts that work from the ability to increase the rate of reaction by decreasing the activation energy of a reaction. (Blanco, Blanco 2017) An enzyme can do this 10^8- to 10^10 fold, sometimes even 10^15 fold. (Malacinsk, Freifelder 1998) The substrate will momentarily bind with the enzyme making the enzyme-substrate complex, of which the shape of the substrate is complimentary to the shape of the active site on the enzyme it is binding with. There are two main theories as to how an enzymes and substrates interact, the lock-and-key model and induced fit theory. The lock-and-key model suggests that the enzyme has a specific shape that fits the substrate and only that substrate. The induced fit theory says the active site and substrate are able to change shape or distort for the reaction to take place with (Cooper,

 produced per minute. The results show a trend wherein increased concentrations of sucrose increase the rate of cellular respiration.