Microorganisms as research tools

Introduction
• Microorganisms are minute living things that are individually too small to be seen with the naked eye. They can only be seen with the help of microscope. They include bacteria, fungi, protists, microscopic plants (green algae); and animals (plankton and the planarian). Some microbiologists also include viruses. Microbiology is the study of microbes. It is divided into two major fields: theoretical and applied. In applied microbiology, industrial and pharmaceutical microbiology are the most promising areas.

Microorganisms in genetic engineering
• Genetic engineering is the manipulation of genes. Also called recombinant DNA technology. Plasmid method, is the most familiar technique in genetic engineering in which pieces of DNA (genes) are introduced into a host by means of a carrier (vector) system. Vector - Plasmid (circular bacterial DNA) Human insulin (used against diabetes), and human growth hormone (used to treat pituitary dwarfism) are the two products commercially produced by this method. E. coli bacterium is used in the production of human insulin.

Antibiotics
• Antibiotics are biochemicals secreted by microorganisms which, in low concentration, inhibit the growth or kill other disease causing microorganisms.

• Out of 6000 known antibiotics, 3000 are obtained from just six genera of filamentous fungi and about 2,000 from three genera of bacteria.

Antibiotics of Fungal Origin
Antibiotic (Product) Ampicillin (a penicillin derivative Methichillin Microbial source Penicillium sp. Mode of action Inhabits cellwall synthesis Inhibits cell Primary spectrum Gram (+) Gram (-) bacteria Staphylococci

Penicillium sp.

Cephalosporin

Cephalosporium Inhabits cellsp. wall synthesis

Gram (+) bacteria

Antibiotics of Bacterial Origin

Antibiotic (Product) Nystatin Bacitracin

Microbial source S. rimosus Bacillus subtilis

Mode of action

Primary spectrum

Interferes with Intestinal Candida, protein synthesis other fungi Inhibits cell wall synthesis Gram-positive bacteria

Vitamin Production
• Riboflavin (vitamin B2) is recovered as a by-product of acetone butanol fermentation however using the yeasts, Eremothecium ashbyii and Ashnua gossypii this vitamin can be produced comercially on a large scale by microbial fermentation Vitamin B12 (Cyanocobalamine; Cobalbumin) is recovered as a byproduct of streptomycin and aureomycin antibiotic fermentations. By microbial fermentation using the bacteria, Propionibacterium shermanii or Pseudomonas denitrificans, the vitamin can be produced on a large-scale.

Biotransformation of Steroids
• Steroids are complex chemical substances containing tetracyclic carbon ring. Steroid hormones regulate various metabolic aspects in man. Microfungi and bacteria has become very important in their production (biotransformation) in pharmaceutical industry. Cortisone is a steroid hormone used to relieve the pain associated with rheumatic arthritis in humans. Chemically synthesized from deoxychloric acid. It completes in 37 different steps, requiring extreme conditions of temperature and pressure for completion and resulting in a very expensive product.

The difficult aspect is the introduction of an oxygen atom at the number 11 position of the steroid ring of progesterone, an intermediate. If progesterone is added to a fermentation tank containing Rhizopus nigricans growing for approximately a day, the progesterone is hydroxylated at the number 11 position of its steroid ring to form a 11-hydroxyprogesterone. The product is recovered by extraction with methylene chloride or various other solvents, purified, and recovered by crystallization. 11-a-hydroxyprogesterone is then subjected to chemical synthesis steps and, finally, cortisone is obtained. The process is easy and has lowered the original cost 400-fold.

Vaccine
• A vaccine is a biological preparation that improves immunity to a particular disease. It typically contains an agent that resembles a disease-causing microorganism, and is often made from weakened or killed forms of the microbe or its toxins. The agent stimulates the body's immune system to recognize the agent as foreign, destroy it, and "remember" it, so that the immune system can more easily recognize and destroy any of these microorganisms that it later encounters. Four basic types of vaccine are: -Inactivated vaccines -Live, Attenuated Vaccines -Subunit Vaccines -Recombinant Vector Vaccines

Bacterial and Viral Vaccines in Current Use
Baterial disease Vaccine Composition

Cholera

Crude fraction of Vibrio cholera

Tuberculosis

Attenuated Mycobacterium tuberculosis Purified polysaccharide from Haemophilus influenze type b

Haemophilus Meningitis

Viral disease

Vaccine Composition

Hepatitis B

Subunit Vaccine

Measles

Attenuated Virus

Microbial enzyme
• Microbial enzymes are most widely used in the food and beverage industries and as protease detergents in washing powders. The most economical and convenient method of producing these enzymes is by microbial fermentation.

Some important microbial enzymes and their applications
Source (genus) Bacillus Enzymes α-Amylase Reaction Application

Starch hydrolysis Converts starch to glucose in food industry Protein digestion Conversion of glucose of fructose Help laundering Production of high fructose syrups

Proteases Streptomyces Glucose isomerase

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