metabolic activities of bacteria

8/18/2019 Metabolic activities of bacteria

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PROTEIN AND AMINO ACID UTILAZATIPHENYLALANINE DEAMINATIONLYSINE DECABOXYLASE)

GROUP 3MEB32

Encabo,Gu!!"#$o, I%#o&


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Raw materials that are found in the environment are used bymicroorganisms in order to grow and multiply

The production of by-products through metabolism can be used inidentication of microorganisms

Their activities in nature and pathogenecity are some important fwhy identication of bacteria is necessary

Dierent tests were conducted in this activity to determine the

bacteria’s metabolic activity

INTRODUCTION


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The types of biochemical reactions each organism undergoesas a "thumbprint" for its identication.

ach dierent species of bacterium has a dierent molecule D!.

#ince D! codes for protein synthesis$ then dierent speciesbacteria must$ by way of their uni%ue D!$ be able to synthesdierent protein en&ymes

This in turn means that dierent species of bacteria must cadierent and uni%ue sets of biochemical reactions.

CONT'


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(' LYSINE DECABOXYLASEPROCEDURE

Lab"!

• 3 tubes• LDC

Inocu!a"

• E. aerogenes• C. Freundii• Unknown bacteria

La*"# • Mineral oil

Incuba"

• 37℃•

48 hrs


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ts !ur!ose is to see i" the #icrobe can use the a#inoacid !*&n" as a source o" carbon and energ% "or growth.

Use o" !*&n" is acco#!lished b% the en&%#e !*&n"+"ca#bo*!a&".

Mineral 'il ( noculated tubes #ust be !rotected "ro# air wi!a*"# o- &"#!" $n"#a! o!. E)!osure to air #a% causealkali&ation at the sur"ace o" the #ediu# which #akes the tein*alid.

LYSINE DECARBOXYLASE


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The medium used is lysine decarboxylase broth. The mediua nutrient broth to which (.)* lysine is added. n importantcomponent of the medium is a modest amount of glucose$necessary for the process to proceed.

+ncreased p, of the medium is detected by color change of theindicators bromcresol purple and cresol red.

, +!D+/T0R 12romcresol turns3

purple at an al4aline p, 51

yellow at an acidic p, -1 p,6).7

LYSINE DECARBOXYLASE

+((-


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Lysine, 0rnithine$ and arginine are the three amino acids routinely testthe identication of Enterobacteriaceae (large family of 8ram-negativbacteria).

#almonella$ scherichia coli$ 9ersinia pestis$ :lebsiella and #higella.

The specic amine products are3

;ysine- /adaverine

0rnithine-utrescine

rginine- /itrulline

These by- products are su


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eagent /ositi*e 0egati*e

LYSINE DECARBOXYLASERESULTS

none /ur!le 1ellow ( orange

2acteria

E. coli

E. aerogenes

C. "reundii

/. *ulgaris

L%sine Utili&ation

,

(

(

(


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LYSINE DECARBOXYLASERESULTS

UUn.no/n2 n"%a0"

E'a"#o%"n"&n"%a0"

E' co!Po&0"

S'au#"u&N"%a0


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2' PHENYLALANINE DEAMINATIONPROCEDURE

Label

• 3 tubes• /

noculate

• E. Coli• /. *ulgaris• Unknown bacteria

ncubate

• 37℃• 48 hrs

"ter 48hrs

• 5 dro!s• Ferric chloride reagent


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#ome bacteria have the ability to deaminate the amino acid

phenylalanine$ as they can produce the en&yme ‘phenylalanindeaminase’.

Deaminase = an en&yme which remove the amine group from tamino acid phenylalanine and releases the amine group as freeammonia !,>1 and phenylpyruvic acid 4et acid1 is produced.

henylpyruvic acid forms green color with ferric chloride so

PHENYLALANINEDEAMINATION


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the test bacterium is grown on agar slants containing

phenylalanine. +f the bacteria have the ability to deaminate phenylalanine$ thcolor of the slants changes from yellow to green$ upon dropping?(* ferric chloride solution after incubation @A hrs.1.

+f the medium remains to its color$ the organism is negative fophenylalanine deaminase production.

PHENYLALANINE DEAMINATIONRESULTS

positive negative


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Bac"#a

E' co!

E' a"#o%"n"&

C' -#"un+

P' 0u!%a#&

P1"n*" #o+

,!9;;!+! 8R,!9;;!+! DB+!# BD+CB1

+t contains nutrients and D;-phenylalanine.+t is used to dierentiate members of the

genera Proteus, Morganella and Providencia from otherEnterobacteriaceae. This test determines whether the microbeproduces the en&yme phenylalaninedeaminase$ which is needed for it to use

the amino acid phenylalanine as a carbonand energy source for growth.

PHENYLALANINE DEAMINATIONRESULTS


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htt!6www.austincc.edu#icrobug&!hen%lalaninedea#inasetest.!h!

htt!6#icrobeonline.co#decarbo)%lation(test(t%!es(uses(!ris(!rocedure(results

htt!6www."rilabo.!t#gs!rodutosi#!ort8854.

9echnicalData:heet.!d"

htt!6www.%ourarticlelibrar%.co#e)!eri#ents!hen%lalaninei&e(test(on(bacteria(to(;nd(out(their(abilit%(to(dea#inate(the(

ino(acid(with(;gure8>

RE5ERENCES
http://www.austincc.edu/microbugz/phenylalanine_deaminase_test.phphttp://www.austincc.edu/microbugz/phenylalanine_deaminase_test.phphttp://microbeonline.com/decarboxylation-test-types-uses-principles-procedure-results/http://microbeonline.com/decarboxylation-test-types-uses-principles-procedure-results/http://microbeonline.com/decarboxylation-test-types-uses-principles-procedure-results/http://www.frilabo.pt/Imgs/produtos/import/88014.TechnicalDataSheet.pdfhttp://www.frilabo.pt/Imgs/produtos/import/88014.TechnicalDataSheet.pdfhttp://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.yourarticlelibrary.com/experiments/phenylalanine-deaminize-test-on-bacteria-to-find-out-their-ability-to-deaminate-the-amino-acid-with-figure/26585/http://www.frilabo.pt/Imgs/produtos/import/88014.TechnicalDataSheet.pdfhttp://www.frilabo.pt/Imgs/produtos/import/88014.TechnicalDataSheet.pdfhttp://microbeonline.com/decarboxylation-test-types-uses-principles-procedure-results/http://microbeonline.com/decarboxylation-test-types-uses-principles-procedure-results/http://microbeonline.com/decarboxylation-test-types-uses-principles-procedure-results/http://www.austincc.edu/microbugz/phenylalanine_deaminase_test.phphttp://www.austincc.edu/microbugz/phenylalanine_deaminase_test.phphttp://www.austincc.edu/microbugz/phenylalanine_deaminase_test.php

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