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Recommended: Enzymes biochemistry
In order to learn even more about my specimen’s metabolic functions, I ran an experiment using a type of differential medium called litmus milk. This differential medium or any other type allows me to actually see certain changes that occur in the tubes after a certain metabolic reaction has taken place (Black, 2015). For this experiment two tubes that contain skin milk and the pH indicator, litmus were inoculated with specimens Ca and Cb. My first litmus milk tube was inoculated with a strain of specimen Ca that was taken from my specimen Ca glucose tube. While my second litmus milk tube contained a strain of specimen Cb that was taken from my specimen Cb lactose tube. After inoculation, both litmus milk tubes were put in an incubator at 37°C …show more content…
While the tube for specimen Cb turned a tannish white in the lower half of the tube while the top stayed the lavender inoculated tube color. Do to this evidence I determined that both specimens Ca and Cb cannot use the process Casein hydrolysis or Casein coagulation due to lack of soft or firm curds in both tubes. Since there was no casein curds formed, I concluded that specimens Ca and Cb also cannot perform the process of proteolysis. My conclusion is supported by the fact that there was no clearing of the medium. I have also determine that neither of my organisms can make the enzymes rennin, proteolytic or even proteases. I know my specimens cannot produce proteases due to the fact that there was no blue coloring in the tubes which means that the byproduct Ammonia was not produced to increase the pH. Since neither of my specimens can make these enzymes, I concluded that my specimens cannot break down lactose or casein. Although I did learn that specimen Cb can reduce litmus due to the evidence that the lower part of the tube turned a tannish white color with a purple ring at the top. This color change from a purple to a white means that the litmus was reduced turning it clear and leaving the white of the milk to show. Finally I know that specimen Ca cannot reduce litmus due to the fact that the tube had no change in
I identified the genus and species of an unknown bacterial culture, #16, and I applied the following knowledge of morphologic, cultural and metabolic characteristics of the unknown microorganism according to the laboratory manual as well as my class notes and power point print outs. I was given an incubated agar slant labeled #16 and a rack of different tests to either examine or perform myself; the tests are as follows: Gram Stain; Nutrient Gelatin Test; Carbohydrate Fermentation; Dextrose, Lactose and Sucrose; IMVIC tests; Citrate, Indole, Mythel-Red and Vogues Proskauer test; as well as a Urease and TSI Test.
We actually saw this change occur by watching the RER values increase. At the 2:30 mark Kerbi was using 100% fats as her fuel source with an RER of .75. One minute later, at an RER of .80 she was using 50% fat and 50% carbohydrates. In between minutes 6:30 with and RER of .99 and less than the 7:00 mark with an RER of 1.04 she was using primarily 100% carbohydrates as her fuel source. Lactate began accumulating in Kerbi’s blood at the 7:00 mark with an RER of 1.04, a speed of 3.4 and a grade of 14. I know this because her RER was above the 100% carbohydrate level of 1.00, and this is also the time at which she felt uncomfortable to the point that the test
Streak plate technique was used to isolate pure culture for each bacteria (2). The Gram stain was used to determine Gram reaction and morphology of each bacteria (2) Selective and differential media such as, salt agar, MacConkey agar and blood agar were used for bacterial identification (2). Gelatin deeps were inoculated to detect production of gelatinase (2). Starch Agar plate were inoculated to detect amylase (2). Ocular reticle used to determine bacteria size (2). Motility deeps were inoculated to detect motility on bacteria (2). Thioglycollate broth used to determine oxygen requirements (2). Carbohydrate fermentation
These labels indicated the lactose solution that was be placed into the mini-microfuge tubes. The varying lactose ph solutions were obtained. The four miniature pipets were then used, (one per solution,) to add 1mL of the solution to the corresponding mini-microfuge tubes. When this step is completed there were two mini-microfuge tubes that matched the paper towel. Then, once all of the solutions contained their respective lactose solutions, 0.5mL of the lactase enzyme suspension was added to the first mini-microfuge tube labeled LPH4 on the paper towel, and 4 on the microfuge tube. As soon as the lactase enzyme suspension was added to the mini-microfuge tube, the timer was started in stopwatch mode (increasing.) When the timer reached 7 minutes and 30 seconds, the glucose test strip was dipped into the created solution in the mini-microfuge tube for 2 seconds (keep timer going, as the timer is also needed for the glucose strip. Once the two seconds had elapsed, the test strip was immediately removed, and the excess solution was wiped gently on the side of the mini-microfuge tube. The timer was continued for 30 addition seconds. Once the timer reached 7:32 (the extra two seconds accounting for the glucose dip), the test strip was then compared the glucose test strip color chart that is found on the side of the glucose test strip
The concentration of Milk Ø The volume of Rennin Ø The volume of Milk Ø The temperature of the reaction Ø Agitation The factor I have chosen to explore is the concentration of Rennin because I believe that varying the temperature of the reaction is very hard to control and therefore may be inaccurate and agitation is a very simple investigation.
Measurement of Ki of a competitive inhibitor (lactose)- For the third part of the experiment, three duplicates of 11 test tubes were set up exactly as in the previous step. The first duplicate was series A while the second duplicate was series B and the last one was series C. Series A did not contained any inhibitors so additional water was added to each tube. Series B contained the 50mM lactose while series C contained the 150mM lactose. The amounts of buffer, ONPG, and water added to each tube in each series are listed in detailed in tables 4, 6, and 8. The Lineweaver- Burk plot of each series was created using the same steps used in the second part of the experiment. Then, the obtained line equation from each plot was used to calculate
The purpose of this experiment was to discover the specificity of the enzyme lactase to a spec...
The independent variable for this experiment is the enzyme concentration, and the range chosen is from 1% to 5% with the measurements of 1, 2, 4, and 5%. The dependant variable to be measured is the absorbance of the absorbance of the solution within a colorimeter, Equipments: Iodine solution: used to test for present of starch - Amylase solution - 1% starch solution - 1 pipette - 3 syringes - 8 test tubes – Stop clock - Water bath at 37oc - Distilled water- colorimeter Method: = == ==
...at keep organisms alive. “Proteins are the most structurally sophisticated molecules known” (Campbell, 1999) which is reason enough to study them. The techniques we learned in this lab form a basis from which a detailed study of proteins is possible. Following our procedure we were successfully able to set up a quantifying assay to determine the amount of protein within a milk sample, although our yield percentage was rather low. However, errors in this lab (in the form of a low yield percentage) may have an origin from our last lab. In the process of extracting proteins from the milk sample, we may have inadvertently lost some of the protein through erroneous measurements, or perhaps through poor handling of either ammonium sulfate or the dialysis tubing. While not sufficient enough (at this point) to invalidate our results, they do explain the major difference between the expected and the actual amount of protein extracted.
After that bolt the slide dry with bibulous paper. After that examine the slide under the oil immersion lens. After determining the Gram stain reaction, 18 specific biochemical tests were performed for further analysis. The way of biochemical test was different but need to incubate at 37 degree Celsius. Phenol red lactose, phenol red dextrose, phenol red sucrose, methyl red, voges-proskauer, citrate utilization test, urea hydrolysis, and nitrate reduction are the media which are in test tube as liquid. Which were use Inoculating loop to deep tube with assigned bacteria. Triple sugar iron, and citrate utilization are in slant and we use Inoculating needle to deep tube with assigned bacteria. Again, starch hydrolysis, casein hydrolysis, lipid hydrolysis, oxidase test and catalase test were test from agar plate which use inoculating loop. For those media, which were in agar plate, inoculate with loop by making a line streak down the center of the agar. After that all 18 medias need to incubate at 37 degree Celsius for 48 hours. After 48 hours, some media show bacteria’s characteristics and morphology without adding extra reagent. Some media need reagent to examine morphology and physiology. Experiment such as MRVP needs methyl red, Barritt’s A, and Barritt’s
Casein Hydrolysis: The skim milk agar plate did not have any clearing around the growth. This is a negative test result.
Pipette 5cm cubed of casein suspension into one boiling tube and 5cm cubed of protease solution into another tube.
The concentration of the following solutions can be measured using a spectrophotometer. In our lab, we will be using the Bradford Assay procedure to measure the concentration of our protein samples. A spectrophotometer is a machine that passes light through a cuvette that can detect how much light a solution can absorb (Absorbance). The absorbance from our measured unknowns can be used to determine the concentration of protein in the solution. If any of our test solutions produce the highest absorptivity, then that solution has the highest concentration of protein because more protein more protein molecules will bind to the Coomassie dye, yielding a dark blue color. If the brand company of muscle milk claims their product have more protein, then we expect to have a high absorptivity and dark blue color for Muscle Milk than the other
If the test was down right, Proteus vulgaris should have be able to breaks down gelatin to make it a liquid (Herter & Ten Broeck, 1911). This means that they desecrate a proteinase enzyme that breaks down gelatin. E. coli should not have been able to break down the gelatin (Exoenzymes). In the urea broth, the Proteus vulgaris broth turned pink and the E. coli did not. This means that the Proteus vulgaris is able to secrete the enzyme urease that breaks down area and forms ammonia making the broth alkaline and pink. Since E. coli did not turn pink, it does not secrete urease.
In our Biology Lab we did a laboratory experiment on fermentation, alcohol fermentation to be exact. Alcohol fermentation is a type of fermentation that produces the alcohol ethanol and CO2. In the experiment we estimated the rate of alcohol fermentation by measuring the rate of CO2 production. Both glycolysis and fermentation consist of a series of chemical reactions, each of which is catalyzed by a specific enzyme. Two of the tables substituted some of the solution glucose for two different types of solutions. They are as followed, Table #5 substituted glucose for sucrose and Table #6 substituted the glucose for pH4. The equation for alcohol fermentation consists of 6 Carbons 12 Hydrogens 6 Oxygen to produce 2 pyruvates plus 2 ATP then finally the final reaction will be 2 CO2 plus Ethanol. In the class our controlled numbers were at Table #1; their table had 15 mL Glucose, 10 mL RO water, and 10 mL of yeast which then they placed in an incubator at 37 degrees Celsius. We each then measured our own table’s fermentation flasks every 15 mins for an hour to compare to Table #1’s controlled numbers. At