METABOLIC PROCESSES AND METABOLITES

No. 6

ETHANOL PRODUCED BY YEAST Saccharomyces cerevisiae Alcoholic beverage CITRIC ACID PRODUCED BY Aspergillus niger Food industry & enhances oil recovery GLUTAMIC ACID AND LYSINE Amino acid produced by Flavor enhancer/ Feed Additive

Corynebacterium glutamicum

POLYSACCHARIDE Xanthomonas spp Food industry, oil recovery

PRIMARY METABOLITES

FUELING REACTIONS Glycolysis, Tricarboxylic acid cycle (TCA) or Krebs cycle, and the Electron Transport System (ETS)

Products of Fueling Reactions Precursor metabolites Metabolic energy (ATP) Reducing power (NADH) C 1 units

SECONDARY METABOLITE

PENICILLIN ANTIBIOTIC CEPHALOSPORIN ANTIBIOTIC STREPTOMYCIN ANTIBIOTIC GRISEOFULVIN ANTIFUNGAL PEPSTATIN TREATMENT OF ULCER CYCLOSPORIN IMMUNOSUPPRESANT GIBBERALLIN PLANT GR OWTH REGULATOR LOVASTATIN CHOLESTEROL SYNTHESIS

INHIBITOR

PCR - Polymerase Chain Reaction is in vitro DNA replication which is a process of making several copies of the

template DNA in a test tube instead of a living cell

How is PCR is used in a DNA-based detection procedure to

test for the presence of disease causing bacteria or viruses ?

Need to have the following:

a. DNA from unknown sample

b. Polymerase enzyme (usually Taq polymerase)

c. Set of PCR primers (that will anneal to target genes

only present in the bacterium or virus being

detected)

d. dNTPs (nucleotides) necessary for PCR e. Buffers that goes with the enzyme (Taq)

1. To use PCR for detection, a gene unique and specific to a bacterial species or a virus to be detected must be

known. The sequence (partial or complete) of that

unique gene must also be known and available.

2. A set of PCR primers that are complimentary to the ends or even to internal fragments of the target gene

will be designed based on known sequence and then

ordered (from a company that synthesize short pieces

of DNAs called oligonucleotides).

3. Polymerase chain reaction will be performed by adding all the components enumerated above and

incubating the samples in eppendorf tubes in a

thermal cycler.

4. If in vitro replication occurs, the target gene is present and therefore the target bacterium or virus is

present

5. No amplification will mean absence of the target gene or the target organism.

Detect the genes that

code for the target

protein in the

cytosol or the gene

for the membrane

protein or proteins

in the appendages

Detect the

genes that

code for the

coat protein

or tail protein

Target protein in

the cytosol,

membrane, or

appendages

Target

regions

of viral

coat protein

or tail region

DNA BASED DETECTION OF

BACTERIA AND VIRUSES

Subject to

agarose gel

electrophoresis

after PCR

Observe DNA in

agarose gel under

UV light

Observe gel profile

or amplified PCR

products

PCR

F

U Lipid Cell inclusion

E Fatty acid

L

I Lipopolysaccharide

N

G Cell ennvelope Cell membrane

Sugars Glycogen Membranes of

cell

organelles

Flagella

Murein

Amino acid Pili Cytoplasm

Protein Ribosome

Cytosol

RNA

GLYCOLYSIS

TCA OR KREBS Polyriibosome Nucleus

ETS Nucleotides Chromosomes

DNA Nucleoid

R

E

A

C

T

I

O

N

S

BIOSYNTHETIC POLYMERIZATION ASSEMBLY

OXALOACETATE CITRATE

GLUTAMIC ACID

OXOGLUTARATE

TCA CYCLE

OXALOACETATE CITRATE

GLUTAMIC ACID

OXOGLUTARATE

GLYCOLYSIS, TCA, AMINO ACID AND SECONDARY METABOLITE

SYNTHESIS

TCA CYCLE