Sunday, June 21, 2009

carbohydrates as an energy source

Use of carbohydrates as an energy source

Glucose is the major energy source in most life forms.

For instance, polysaccharides are broken down into their monomers glycogen phosphorylase removes glucose residues from glycogen). Disaccharides like lactose or sucrose are cleaved into their two component monosaccharides.

Glycolysis (anaerobic)
Glucose is mainly metabolized by a very important and ancient ten-step pathway called glycolysis, the net result of which is to break down one molecule of glucose into two molecules of pyruvate; this also produces a net two molecules of ATP, the energy currency of cells, along with two reducing equivalents in the form of converting NAD+ to NADH.

This does not require oxygen; if no oxygen is available (or the cell cannot use oxygen), the NAD is restored by converting the pyruvate to lactate (lactic acid) (e. g. in humans) or to ethanol plus carbon dioxide (e. g. in yeast).

Other monosaccharides like galactose and fructose can be converted into intermediates of the glycolytic pathway.

Aerobic
In aerobic cells with sufficient oxygen, like most human cells, the pyruvate is further metabolized. It is irreversibly converted to acetyl-CoA, giving off one carbon atom as the waste product carbon dioxide, generating another reducing equivalent as NADH.

The two molecules acetyl-CoA (from one molecule of glucose) then enter the citric acid cycle, producing two more molecules of ATP, six more NADH molecules and two reduced (ubi)quinones (via FADH2 as enzyme-bound cofactor), and releasing the remaining carbon atoms as carbon dioxide.

The produced NADH and quinol molecules then feed into the enzyme complexes of the respiratory chain, an electron transport system transferring the electrons ultimately to oxygen and conserving the released energy in the form of a proton gradient over a membrane (inner mitochondrial membrane in eukaryotes).

Thereby, oxygen is reduced to water and the original electron acceptors NAD+ and quinone are regenerated. This is why humans breathe in oxygen and breathe out carbon dioxide. The energy released from transferring the electrons from high-energy states in NADH and quinol is conserved first as proton gradient and converted to ATP via ATP synthase.

This generates an additional 28 molecules of ATP (24 from the 8 NADH + 4 from the 2 quinols), totaling to 32 molecules of ATP conserved per degraded glucose (two from glycolysis + two from the citrate cycle). It is clear that using oxygen to completely oxidize glucose provides an organism with far more energy than any oxygen-independent metabolic feature, and this is thought to be the reason why complex life appeared only after Earth's atmosphere accumulated large amounts of oxygen.

Source: http://en.wikipedia.org/wiki/Biochemical



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Saturday, June 20, 2009

Selective & Differential Media

Selective & Differential Media


What is selective media?

Selective media, is made up of certain nutrients that can either inhibit or enhance the growth of certain bacteria. This is why they call it selective media, it is generally only selective for one kind of bacteria or does certain things when that bacteria or fungus is present.

Some examples of selective media include:
* blood agar (used in strep tests), which contains bouvine heart blood that becomes transparent in the presence of hemolytic Streptococcus
* MacConkey agar for Gram-negative bacteria
* Hektoen enteric agar (HE) which is selective for Gram-negative bacteria
* mannitol salt agar (MSA) which is selective for Gram-positive bacteria and differential for mannitol



How about Differential Media?

Differential media or indicator media distinguish one microorganism type from another growing on the same media. This type of media uses the biochemical characteristics of a microorganism growing in the presence of specific nutrients or indicators (such as neutral red, phenol red, eosin y, or methylene blue) added to the medium to visibly indicate the defining characteristics of a microorganism. This type of media is used for the detection of microorganisms and by molecular biologists to detect recombinant strains of bacteria.

Examples of differential media include:
* MacConkey (MCK), which is differential for lactose fermentation
* mannitol salt agar (MSA), which is differential for mannitol fermentation

A more illustration on individual agar;
Macconkeys agar
Mannitol salt agar

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Thursday, June 18, 2009

Oxidase Test

Oxidase Test NOTE: This test is for Gram-negative organisms.The oxidase test determines whether a microbe can oxidize certain aromatic amines, for example, p -aminodimethylaniline, to form colored end products. This oxidation correlates with the cytochrome oxidase activity of some bacteria, including the genera Pseudomonas and Neisseria. While a positive oxidase test is important in the identification of these genera, the test is also useful in characterizing the enteric bacteria ( Enterobacteriaceae), which are oxidase-negative.

Modified Oxidase Test (Microdase Disks) NOTE: This test is for Gram-positive, catalase-positive cocci, only. You must use bacteria from a culture grown on Blood agar for 24 - 36 hours. Cultures which are too young or too old may give inaccurate results.
This test is used for differentiating Micrococcus from Staphylococcus . Micrococci should yield a positive result. Staphylococci should yield a negative result, with the exception of Staphylococcus sciuri.


Enzymatic activities are used to differentiate bacteria.
Some bacteria contain enzymes that help to speed up the rate of chemical reactions. One of these reactions is the breaking down of compounds.
Enzyme that adds oxygen during the reaction of breaking down compound is also known as oxidase.


To test for oxidase, an oxidase test is being used.
An oxidase test is a test to determine whether a bacterium contain an enzyme cytochrome oxidase. Cytochrome oxidase takes part in the electron transport chain by transferring electrons to oxygen from a donor molecule.
The oxidase reagent contains a chromogenic reducing agent, Tetramethyl-p-phenylenediamine (TMPD) or Dimethyl-p-phenylenediamine (DMPD), which is a compound that changes color when it becomes oxidized and remain colorless when reduced.


If bacterium contains cytochrome oxidase, it can use the oxygen for energy production with an electron transfer chain. The oxidase reagent will turn blue or purple within a short time. The reaction is positive.

If the bacterium does not contain cytochrome oxidase, the reagent will remain colorless. The reaction is negative.


Bacteria that show positive reactions (contain cytochrome oxidase) are oxidase positive. Examples of such bacteria are preliminary identification of Neisseria and Moraxella genera.
Bacteria that do not show positive reactions (do not contain cytochrome oxidase) are oxidase negative. Examples of such bacteria are all enteric bacteria.

Open link to view
Catalase reaction




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Sources:
http://www.tgw1916.net/video_pages/oxidase.html

http://web.fccj.edu/~lnorman/images/oxidase%20test.jpg


Sources:
http://en.wikipedia.org/wiki/Oxidase_test
http://www.austincc.edu/microbugz/oxidase_test.php
http://web.mst.edu/~microbio/Lab_Supplement/Oxidase.html
http://www.tgw1916.net/video_pages/oxidase.html