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What is fermentation - Being able to grow anaerobically in the presence of a suitable organic compound, such as a sugar. Several reduced products are excreted by the cell as a way of disposing of the electrons derived from oxidation. Function and Types of hydrolytic enzymes - Hydrolytic enzymes...

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MCB 2610 Lab Practical
What is fermentation - Being able to grow anaerobically in the presence of a suitable organic
compound, such as a sugar.

Several reduced products are excreted by the cell as a way of disposing of the electrons derived from
oxidation.

Function and Types of hydrolytic enzymes - Hydrolytic enzymes hydrolyze (break down) complex
molecules to get food that can be transported into the cell.

1. Amylase

2. Caseinase

3. Gelatinase

Amylase - catalyzes starch hydrolysis. Reacts readily with iodine to produce a dark blue color.

Clear zone indicates amylase production.

Iodine makes the starch blue.

Caseinase - Enzymes that hydrolyze milk protein (casein) into peptides and ultimately into amino
acids

Clear zone where caseinase is digested

Catalase - An enzyme which protects cells from hydrogen peroxide - a highly reactive and toxic
chemical produced as a byproduct of oxygen metabolism

H2O2 in the presence of catalase will produce water and oxygen as a byproduct

If catalase positive, bubbles of O2 will form when a drop of H2O2 is placed on the organism

Complex media - Exact chemical composition is not known

Most are made from inexpensive sources

Diverse mixture of proteins (peptides), sugars, vitamins, minerals

, Death phase - Occurs if conditions remain unchanged (no new medium or environment remains
stable). The cell death rate will eventually exceed any remaining cell division.

Defined media - Made up of a strict list of compounds

Forces organisms to synthesize necessary amino acids, nucleotides, lipids, polysaccharides, vitamins

Allows scientists to characterize the versatility and limitations of an organism's metabolic capacity

Defined media can be used to help define the specific requirements for a particular organism

Differential media - Allows certain bacteria to be differentiated from others, typically by changing
of color, production of a zone of clearing etc.

Dye used for spore staining - 5% aqueous Malachite green dye

Gelatinase - An enzyme that hydrolyzes gelatin, a protein derived from collagen.

If gelatin is hydrolyzed, the media will become liquid

General purpose media - WIll support growth of a wide range of organism

Used to grow up dense cultures of bacteria, typically for experimentation

How does SF select for streptococci - Streptococci lack a respiratory system, so they use
fermentation. Brom cresol purple is an acid indicator that changes color when acid is produced,
indicating fermentation of the glucose.

Purple = negative for fermentation of glucose

Yellow = positive for fermentation of glucose

How is bacterial growth measured? - Using a spectrophotometer, you can measure the optical
density (absorbance) over a period of time

How is microbial growth defined? - Increase in population size

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