BIO 3124 Lecture Notes - Lecture 9: Microaerophile, Anaerobic Respiration, Cellular Respiration

BIO 3124 Lecture Notes - Lecture 9: Microaerophile, Anaerobic Respiration, Cellular Respiration

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Greater nutritional complexity: more substances need to be provided for survival. Lower nutritional complexity = higher biosynthetic capacity: can make more things on your own, ex. Plants have a greater biosynthetic capacity and lower nutritional complexity than humans. The greater the biosynthetic capacity, the lower the nutritional requirements. Composed of rich and complex ingredients (ex. soya protein extract, milk protein extracts, blood products, tomato juice, etc. ) Exact chemical composition is unknown (ex. blood today and blood tomorrow is not identical) Medium favourable for organisms with a higher nutritional requirement. Can contain up to 80 different ingredients. Allows the growth of a restricted number of microorganisms - tailor-made for a given microorganisms nutritional requirement. Composition is highly variable according to the microorganism. Contain compounds which inhibit or kills the undesired organisms. Medium containing penicillin only allows the growth of penicillin resistant microorganisms. Often contain ph indicators - allows to discriminate different metabolisms.

BACTERIAL GROWTH AND CONTROLLING BACTERIALGROWTH

1. Based upon your results, is your bacteria sample a strictaerobe, strict anaerobe, facultative anaerobe, or microaerophile?Why? â Look at your results from OF Glucose and OF Glucose with oiltest.

2. Did the bacteria grow in the tryptone or malonate media? Whyor why not? â Remember, you can tell if they grew if culture wasturbid (cloudy).

3. For the media used in this portion of the experiment, are themedia complex or synthetic? Are the media selective, differential,both, or neither? Identify the carbon and nitrogen source for eachmedium. â Read more about what these terms mean starting on page178 in your book right below Figure 6.11. Look up each media withthe M? button. Remember, nutrient broth is a complex media, mineralsalts is a defined media.

4. How many possible bacteria were remaining after conductingthese biochemical tests?

5. Using the information in the Identification matrix, whatadditional tests might help you to identify the bacteria in yourcase study? âLook at the View Identification Matrix under the Viewmenu. What tests would you perform? Look for ones that have a mixof + and â because they would help distinguish betweenorganisms.

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1. The pH for the water in the MSA recipe is expected to be from 7.2-7.6. If the pH was too low, what could you expect to see? Select one:

a. The entire plate would be yellow

b. Nothing abnormal

c. The agar would crack at pH 6.5

d. E. coli would turn purple

2. Which of the following would the MSA medium NOT be considered: Select one:

a. Complex medium

b. Chemically defined medium

c. Differential medium

d. Selective medium

3. If you took a swab from a patient and cultured it on a MSA plate, which of the following could be identified by just using this test? Select one:

a. Staphylococcus epidermidis

b. Escherichia coli

c. Staphylococcus aureus

d. Micrococcus luteus

4. If you streaked the 3 organisms in this lab on a PEA plate and after incubating for 2 days at 37 degrees Celcius nothing grew (but they did grow on a NA plate made at the same time and grown in the same conditions), which of the following is the most likely cause for this result? Select one:

a. The agar was toxic.

b. The beef extract was bad.

c. The temperature was too low.

d. Too much phenylethyl alcohol was used

. e. The temperature was too high.

5. Why is the NA plate inoculated in addition to the Columbia CNA with 5% sheep blood agar plate? Select one:

a. Used to compare growth on non-selective medium

b. Used as second selective medium

c. Used to promote hemolysis

d. Used because it is easier to see

e. Used as a practice plate

Enzymes Lab Report

Biology 1408

Temperature and Enzyme Activity

Based on Lab, Section III, Exercise 3 record the relative colorintensity at five-minute intervals to determine the temperaturerange for catechol oxidase.

Construct a bar graph of the data for the ten-minute readings.Label axes and provide units. Sign, date and prepare an image ofyour graph and include it with this lab report.

What is the optimal temperature range for catechol oxidaseactivity?

pH and Enzyme Activity

Within what pH range will catechol oxidase catalyze catecholand oxygen to form benzoquinone?

4-12

Define pH.

What is the role of catechol in the experiment?

Which test tube contains the most acidic buffer solution?

Cofactors and Enzyme Activity

What is a cofactor?

What cofactor is necessary for catechol oxidase activity?

Summary Questions

What was the role of catechol oxidase in the experiments?

What was the purpose of developing a color intensityscale?

Identify three factors that influence the rate of enzymaticreactions.

Examine the data in Exercise 3 and suggest a hypothesisexplaining why people with a fever feel tired and listless.

With high fever, enzymestructure changes and no longer act as a catalyst

People who have been suddenly submerged in cold water andremained for long periods of time have survived. Examine the datain Exercise 3 and explain how this is possible.

Low temperature ranges willinhabit the reaction of the speed of chemical reaction incells

How does pH range affect the function and structure of catecholoxidase?

Help the enzymes functionbetter

The stomach maintains a pH of approximately 3 to 5. Do allenzymes function at this pH level? Examine the data in Exercise 4and determine if catechol oxidase would function in the stomach.

No, most enzymes functionbetter at ph 6-8

What is enzyme specificity? Give an example.

A specific substrate fits aspecific enzyme.

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Time

(min)

Tube 1

4^oC

Tube 2

23^oC

Tube 3

40^oC

Tube 4

60^oC

Tube 5

80^oC

Tube 6

100^oC