Biochemicals Flashcards
METHYL RED (MR) TEST
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- An organism’s ability to produce acid end products from glucose fermentation
- Glucose
- Mixed acids
- Methyl Red Indicator
- NONE
- Mixed Acid fermentation
- Positive: Red
Negative: Yellow (no color change)
VOGES-PROSKAUER (VP) TEST
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- To detect acetoin, a neutral end product, from glucose fermentation
- Glucose
- Acetoin
- KOH and Alpha naphthol
- NONE
- Butylene Glycol fermentation
- Positive: Red/pink (upper layer of media)
Negative: Yellow (no color change)
SODIUM CITRATE TEST
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- An organism’s ability to use citrate as its sole source of carbon
- Citrate
- NONE
- NONE
- NONE
- NONE
- Positive: Blue
Negative: Green (no color change)
PHENYLALANINE DEAMINASE
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- An organism’s ability to oxidatively deaminate phenylalanine to phenylpyruvic acid (remove an amine group from an amino acid)
- Phenylalanine
- Phenylpyruvic acid
- Ferric chloride (FeCl3)
- Phenylalanine deaminase
- NONE
- Positive: Dark green slant (fades after a few minutes)
Negative: Yellow (no color change - from reagent)
INDOLE TEST - EHRLICH’S PROCEDURE
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- Differences from Kovac’s Procedure??
** Used specifically for??
*** Used specifically for (Kovac’s Procedure)??
- An organism’s ability to split indole from tryptophan
- Tryptophan
- Indole
- Kovac’s (dimethylaminobenzaldehyde)
- Tryptophanase
- NONE
- Positive: Red (middle layer in Ehrlich’s’ top layer in Kovac’s)
Negative: Yellow (no color change)
Extraction of indole with xylene, chloroform, or ether prior to
reagent
** Nonfermenting GNRs and anaerobes
** Enterobacteriaceae
MOTILITY
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- To determine if an organism is motile (flagellated) or non motile (nonflagellated)
- None
- None
- None
- None
- None
- Positive: Hazy appearance/stab line not evident/red color diffused (tetrazolium salts added)
Negative: No hazy appearance/stab line clearly seen/red color localized to stab line (tetrazolium salts added)
UREASE TEST
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
*Media name??
- An organism’s ability to split urea by urease
- Urea
- Ammonia
- None
- Urease
- None
- Positive: Pink
Negative: Yellow (no color change)
- Christensen’s
DECARBOXYLASE MEDIA (LYSINE)
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
*Media name??
- Purpose/Principle: To detect the presence of decarboxylase enzymes in the breakdown of amino acids resulting in amine + CO2
- Substrate: Lysine
- Product(s): Cadaverine + CO2
- Reagent: NONE
- Enzymes: Lysine decarboxylase
- Pathway(s): NONE
- Results: Positive: Purple/Brown
Negative: Yellow
- Moeller’s
DECARBOXYLASE MEDIA (ORNITHINE)
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
*Media name??
- Purpose/Principle: To detect the presence of decarboxylase enzymes in the breakdown of amino acids resulting in amine + CO2
- Substrate: Ornithine
- Product(s): Putrescine + CO2
- Reagent: NONE
- Enzymes: Ornithine decarboxylase
- Pathway(s): NONE
- Results: Positive: Purple/Brown
Negative: Yellow
- Moeller’s
ARGININE DIHYDROLASE
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
*Media name??
- Purpose/Principle: To detect the presence of arginine dihydrolase enzymes in the breakdown of the amino acid, arginine
- Substrate: Arginine
- Product(s): Ornithine
- Reagent: NONE
- Enzymes: Arginine Dehydrolase
- Pathway(s): NONE
- Results: Positive: Purple/Brown
Negative: Yellow
- Moeller’s
KLIGLER IRON AGAR (KIA)
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
*Triple Sugar Iron Agar (TSI) difference??
- An organism’s ability to ferment lactose and/or glucose, with/without the production of gas – the production of hydrogen sulfide is also determined
- Glucose, Lactose, Sodium thiosulfate
- Acid/Basic products, H2S (organism-dependent)
- NONE
- NONE
- NONE
- A/A = glucose and lactose fermentation
K/A = glucose only/no lactose fermentation
K/K = no glucose fermentation
Bubbles = CO2 gas production
Black ppt. = H2S production
*Contains 1.0% sucrose, as well as glucose (0.1%) and lactose (1.0%) – when yellow on the slant, could be from lactose or sucrose utilization
DECARBOXYLASE BASE
Purpose?
Results?
To ensure organism is producing acid by glucose fermentation – used as a control tube for the LOA
Yellow color required for valid interpretation of LOA series
1% CARBOHYDRATES
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- An organism’s ability to degrade specific carbs with the production of acids, with/without visible gas
- Various carbohydrates
- Acid products (organism-dependent)
- None
- None
- None
- Positive: Yellow
Negative: Purple (no color change)
OXIDATION/FERMENTATION (OF) MEDIA
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- To determine an organism’s mechanism for utilization of glucose; fermentative, oxidative or neither
- Glucose (or other carbs)
- Acid products (organism-dependent)
- None
- None
- None
- Positive: Yellow in oiled tube (fermenter)
Yellow in unoiled tube (oxidizer)
Negative: Green (no color change) – asaccharolytic (non-
oxidizer)
CYTOCHROME OXIDASE
- Purpose/Principle:
- Substrate:
- Product(s):
- Reagent:
- Enzymes:
- Pathway(s):
- Results:
- To detect the presence of oxidized cytochrome C
- 4.1% tetramethyl-p-phenylenediamine dihydrochloride or
Dimethyl-p-phenylendiamine + alpha napthol - Cytochrome oxidase
- None
- Positive: Blue
Negative: No color change