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PROTEIN AND AMINO ACID UTILAZATION

PHENYLALANINE DEAMINATION
LYSINE DECABOXYLASE)

GROUP 3
MEB32
Encabo,Guillermo, Igros

INTRODUCTION
Raw materials that are found in the environment are used by
microorganisms in order to grow and multiply
The production of by-products through metabolism can be used in the
identification of microorganisms
Their activities in nature and pathogenecity are some important factors
why identification of bacteria is necessary
Different tests were conducted in this activity to determine the
bacterias metabolic activity

CONT.
The types of biochemical reactions each organism undergoes act
as a "thumbprint" for its identification.
Each different species of bacterium has a different molecule of
DNA.
Since DNA codes for protein synthesis, then different species of
bacteria must, by way of their unique DNA, be able to synthesize
different protein enzymes
This in turn means that different species of bacteria must carry out
different and unique sets of biochemical reactions.

1. LYSINE DECABOXYLASE
PROCEDURE
3 tubes
Label LDC
E. aerogenes
C. Freundii
Inoculat
Inoculat
e
Unknown bacteria

Layer

Mineral oil

37
Incubate
Incubate 48 hrs

LYSINE DECARBOXYLASE
Its purpose is to see if the microbe can use the amino
acidlysineas a source of carbon and energy for growth.
Use oflysineis accomplished by the enzymelysine
decarboxylase.
Mineral Oil - Inoculated tubes must be protected from air with a
layer of sterile mineral oil. Exposure to air may cause
alkalization at the surface of the medium which makes the test
invalid.

LYSINE DECARBOXYLASE

The medium used islysine decarboxylase broth.The medium is


a nutrient broth to which 0.5%lysineis added. An important
component of the medium is a modest amount of glucose,
necessary for the process to proceed.
Increased pH of the medium is detected by color change of thepH
indicators bromcresol purple and cresol red.
(PH INDICATOR )Bromcresol turns:

purple at an alkaline pH (+)


yellow at an acidic pH (-) pH<5.2
(--)

(+)

LYSINE DECARBOXYLASE
Lysine, Ornithine, and arginine are the three amino acids routinely tested in
the identification of Enterobacteriaceae (large family of Gram-negative
bacteria).
Salmonella, Escherichia coli, Yersinia pestis, Klebsiella and Shigella.
The specific amine products are:
Lysine- Cadaverine
Ornithine-Putrescine
Arginine- Citrulline
These by- products are sufficient to raise the pH of the media so that the broth
turns purple.

LYSINE DECARBOXYLASE
RESULTS

Reagent

Positive

Negative

none

Purple

Yellow - orange

Bacteria

Lysine Utilization

E. coli

E. aerogenes

C. freundii

P. vulgaris

LYSINE DECARBOXYLASE
RESULTS

S.
E. coli
aureus Positive
Negativ

E.
aerogenes
negative

Unknown
2 negative

Unknown 3
negative

2. PHENYLALANINE DEAMINATION
PROCEDURE
3 tubes
PA
Label
E. Coli
Inoculat P. vulgaris
Unknown bacteria
e
37
Incubat 48 hrs
e
10 drops
After 48 Ferric chloride reagent
hrs

PHENYLALANINE
DEAMINATION

Some bacteria have the ability to deaminate the amino acid


phenylalanine, as they can produce the enzyme phenylalanine
deaminase.
Deaminase an enzyme which remove the amine group from the
amino acid phenylalanine and releases the amine group as free
ammonia (NH3) and phenylpyruvic acid (ket acid) is produced.
Phenylpyruvic acid forms green color with ferric chloride solution.

PHENYLALANINE DEAMINATION
RESULTS
the test bacterium is grown on agar slants containing
phenylalanine.
If the bacteria have the ability to deaminate phenylalanine, the
color of the slants changes from yellow to green, upon dropping of
10% ferric chloride solution after incubation (48 hrs.).
If the medium remains to its color, the organism is negative for
phenylalanine deaminase production.

positive

negative

PHENYLALANINE DEAMINATION
RESULTS
PHENYLALANINE AGAR
(PHENYLALANINE DEAMINASE MEDIUM)
It contains nutrients and DLphenylalanine.
It is used to differentiate members of the
generaProteus, Morganella
andProvidenciafrom other
Enterobacteriaceae.
This test determines whether the microbe
produces the enzymephenylalanine
deaminase, which is needed for it to use
the amino acidphenylalanineas a carbon
and energy source for growth.

Bacteria

Phenylalanin
e production

E. coli

E. aerogenes

C. freundii

P. vulgaris

REFERENCES

http://www.austincc.edu/microbugz/
phenylalanine_deaminase_test.php
http://microbeonline.com/decarboxylation-test-types-uses-principle
s-procedure-results
/
http://www.frilabo.pt/Imgs/produtos/import/88014.
TechnicalDataSheet.pdf
http://www.yourarticlelibrary.com/experiments/phenylalanine-deamin
ize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-am
ino-acid-with-figure/26585
/