Microorganisms as research tools

ASSIGNMENT ON Microorganisms as Research Tools

Course name: Basic Microbiology Course code: PHRM 202 Semester: Summer 2011

SUBMITTED TO: Abu Taiab Md. Jamaluddin Senior Lecturer Department of Pharmacy East West University SUBMITTED BY: Samiya Khondaker Rinta (ID: 2010-3-70-048) Nabeela Zaman (ID: 2010-3-70-043) Sharmin Sheikh (ID: 2010-3-70-007) Priyata Dey (ID: 2010-3-70-042) Debasree Paul (ID: 2010-3-70-036)

Submission date: 20th March, 2011

Microorganisms as research tools

Abstract Microorganisms or microbes are very important in our lives and are used as tools for research. Microbiology is the study of microbes and it is one of the most applied of all the biological sciences. Microbiology is not a single subject. It has many areas of specialization. It is a science based on the use of pure cultures - looking at one sort of organism at a time. It can be divided into two major fields: theoretical and applied. The major fields of applied microbiology are: Medical microbiology, Aquatic microbiology, Aero-microbiology, Agricultural, Industrial microbiology, Exo- microbiology and Geochemical microbiology. Industrial and pharmaceutical microbiology are the most promising areas of applied microbiology. Many of the techniques developed by microbiologists and researchers are now used in molecular and cell biology to provide the basis for studying higher organisms. Research on microorganisms has enabled us to derive health care products, such as antibiotics, hormones, vitamins, and vaccines, from microbial biosynthesis. Microorganisms also play a central role in recombinant DNA technology and act as the agents for genetic modification. Applying knowledge gained from these techniques can lead to many improvements in the quality of our lives. Keywords: Microorganisms, applied microbiology, microbial biosynthesis, recombinant DNA technology.

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. Microorganisms are very diverse; they include bacteria, fungi, and protists; microscopic plants (green algae); and animals such as plankton and the planarian. Some microbiologists also include viruses, but others consider these as non-living. Microbes are useful tools in research because of their rapid life cycle, their simple growth requirements, and their small size. Microorganisms live in all parts of the biosphere where there is liquid water, including soil, hot springs, on the ocean floor, high in the atmosphere and deep inside rocks within the Earth's crust. Microbes are also exploited by people in biotechnology, both in traditional food and beverage preparation, and in modern technologies based on genetic engineering. Researchers in microbiology, medicine, genetic engineering, and other fields discussed ways to build on and extend successes in microbiology. Microbes fulfill many functions that make life on Earth possible. They are the engines behind the global biogeochemical cycles that release oxygen and absorb greenhouse gases in the atmosphere. They recycle dead material into useful nutrients for new growth. They influence food webs, bioenergy production, waste management and treatment, food production, and symbiotic nitrogen fixation for plants. Because microbes play so many roles in so many environments, they hold incredible potential for industry, agriculture, and medicine. Microorganisms in genetic engineering Genetic engineering alters the genetic makeup of an organism using techniques that introduce heritable material prepared outside the organism either directly into the host or into a cell that is then fused or hybridized with the host (The European Parliament, 2001). It is one of the fields of biotechnology. It is also called recombinant DNA technology. In genetic engineering, the plasmid method, is the most familiar technique and is generally used for altering microorganisms such as bacteria. In this method pieces of DNA (genes) are introduced into a host by means of a carrier (vector) system. Here the vector used is the circular bacterial DNA called the plasmid which, after it acquires the gene of interest is inserted into a bacterial cell. The bacterial cell, containing the foreign DNA, is the host cell and it becomes a permanent feature of the host, being replicated and passed on to daughter cells along with the rest of its DNA. Bacterial

Microorganisms as research tools

cells are transformed and used in production of commercially important products. The examples are production of human insulin (used against diabetes), and human growth hormone (somatotrophin used to treat pituitary dwarfism). Insulin is a simple protein normally produced by the pancreas. In people with diabetes, the pancreas is damaged and cannot produce insulin. Since insulin is vital to the body's processing of glucose, this is a serious problem. Many diabetics, therefore, must inject insulin into their bodies daily. Prior to the 1980s, insulin for diabetics came from pigs and was very expensive. To create insulin inexpensively, the gene that produces human insulin was added to the genes in a normal E. coli bacterium. Once the gene was in place, the normal cellular machinery produced it just like any other enzyme. By culturing large quantities of the modified bacteria and then killing and opening them, the insulin could be extracted, purified and used very inexpensively. Antibiotics produced using microorganisms In medical microbiology microorganisms are used particularly in the production of antibiotics and it has been the greatest boon to humans and other animals. Antibiotics are biochemicals secreted by microorganisms which, in low concentration, inhibit the growth or kill other disease causing microorganisms. Although some 6,000 antibiotics are known to exist, only about 100 are marketed. About 200 new antibiotics are discovered every year by scientists and researchers. However, out of 6,000 antibiotics, about 1,000 have been obtained from just six genera of filamentous fungi (e.g., Penicillium, Cephalosporium) and about 2,000 from just three genera of bacteria (e.g., Streptomyces, Bacillus). Table 1: Fungal metabolic products useful as antibiotics Antibiotic (Product) Ampicillin (a penicillin derivative Cephalosporin Cephalosporium sp. Inhabits cell-wall synthesis Penicillium sp. Inhabits cell-wall synthesis Gram (+) Gram (-) bacteria Gram (+) bacteria Microbial source Mode of action Primary spectrum

Microorganisms as research tools

Methichillin Griseofulvin

Penicillium sp. P. griseofulvum

Inhibits cell

Staphylococci

interferes with cell wall and Pathogenic fungi nucleic acid synthesis

Table 2: Antibiotics of Bacterial Origin (only Streptomyces) Antibiotic (Product) Microbial source Amphotericin B Streptomyces nodosus Interferes with membrane function Chlorotetracycline (Aureomycin) Erythromycin S. erythraeus S.aureofaciens Interferes with protein synthesis Interferes with protein synthesis Neomycin S fradiae Induces abnormal Gram- positive and Gram-negative protein synthesis bacteria used to treat tuberculosis when causative agent becomes resistant against other antibiotics. Novabiocin S. griseus S. niveus S. spheroids S. noursei Nystatin S. rimosus Interferes with protein synthesis Oxytetracycline (Terramycin) Vancomycin Viomycin S. floridae S. orientailis Inhibits cell-wall Broad spectrum Gram-positive bacteria. synthesis Interferes with protein synthesis Neisseria, Clostridium tetani Mycobacterium tuberculosis Intestinal Candida, other fungi Damages cell membrane Gram-positive bacteria Rickettsias, gram-positive bacteria Broad spectrum Fungi causing diseases called mycoses. Mode of action Primary spectrum

Microorganisms as research tools

Table 3: Antibiotics of Bacterial Origin (other than Streptomyces) Antibiotic (Product) Microbial source Bacitracin Bacillus subtilis Mode of action Inhibits cell wall synthesis Polymixin B B. polynixa cell membrane deterioration Primary spectrum Gram-positive bacteria Gram-positive bacteria

Vitamin Production Out of all vitamins now available commercially, vitamin B12 and vitamin B2 (riboflavin) are mainly produced by microbial fermentations. Riboflavin (vitamin B2) is recovered as a by-product of acetone butanol fermentation and is produced by various Clostridium species. But, there are Eremothecium ashbyii and Ashnua gossypii, the yeasts that are used to commercially produce this vitamin by direct fermentation. Vitamin B12 (Cyanocobalamine; Cobalbumin) is recovered as a by-product of streptomycin and aureomycin antibiotic fermentations. Vitamin B12 (Cyanocobalamine) is also produced on large scale by direct fermentation using the bacteria Propionibacterium shermanii or Pseudomonas denitrificans. Biotransformation of Steroids Steroids are complex chemical substances containing tetracyclic carbon ring. They are important therapeutic agents as steroid hormones regulate various metabolic aspects in man (including animals). The chemical synthesis of steroids is very complex and expensive and, therefore, use of microorganisms such as 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. This steroid is chemically synthesized from deoxychloric acid; the chemical synthesis

Microorganisms as research tools

completes in 37 different steps, many of which require extreme conditions of temperature and pressure for completion and the resulting product is very expensive. The most difficult aspect of the chemical synthesis of cortisone is the introduction of an oxygen atom at the number 11 position of the steroid ring of progesterone, an intermediate. When 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 from 11-ahydroxyprogesterone. The product is recovered by extraction with methylene chloride or various other solvents, purified chromatographically, and recovered by crystallization. 11-ahydroxyprogesterone is now subjected to chemical synthesis steps and, finally, cortisone is obtained. The use of biotransformation process to obtain cortisone has made the whole process easy and has lowered the original cost 400-fold. Vaccine A vaccine is a biological preparation that improves immunity to a particular disease. A vaccine 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. The four basic types of vaccine in use today (PM., 1996). These are as follows: 1) Inactivated vaccines: These are produced by killing the disease-causing microorganism with chemicals or heat. 2) Live, Attenuated Vaccines: To make a live, attenuated vaccine, the disease-causing organism is grown under special laboratory conditions that cause it to lose its virulence, or disease-causing properties. 3) Subunit Vaccines: Sometimes vaccines developed from antigenic fragments are able to evoke an immune response. Subunit vaccines can be made by taking apart the actual microbe, or they can be made in the laboratory using genetic engineering techniques. 4) Recombinant Vector Vaccines: A vaccine vector, or carrier, is a weakened virus or bacterium into which harmless genetic material from another disease-causing organism can be inserted.

Microorganisms as research tools

Table 4: Bacterial and Viral Vaccines in Current Use Disease Vaccine Composition Bacterial Cholera Tuberculosis Haemophilus Meningitis Meningococcal meningitis Crude fraction of Vibrio cholera Attenuated Mycobacterium tuberculosis ( BCG) Purified polysaccharide from Haemophilus influenze type b Purified polysaccharide from types A and C

Pertussis ( Whooping Cough) Killed Bordetella Capsular Pneumonia ( by Streptococcus Pneumoniae) Mixture of 23 different capsular Viral Hepatitis B Measles Mumps Polio Rubella Subunit Vaccine Attenuated Virus Attenuated Virus Attenuated Virus OR Inactivated Virus Attenuated Virus

Microbial enzymes Microbial enzymes are most widely used in the food industries and as protease detergents in washing powders. The most economical and convenient method of producing these enzymes is by microbial fermentation (Stanley, 1998). Table 5: Some important microbial enzymes and their applications Source (genus) Bacillus Enzymes -Amylase Reaction Starch hydrolysis Application Converts starch to

Microorganisms as research tools

glucose in food industry Proteases Aspergillus Amyloglucosidase -Galactosidase Protein digestion Dextrin hydrolysis Lactose hydrolysis Help laundering Glucose production Lactose hydrolysis in milk or whey Streptomyces Glucose isomerase Conversion of glucose Production of high of fructose fructose syrups

Thus all these show that research on microbes has numerous significant applications for human welfare. In the future, there will be a need for more rapid diagnoses, increased standardization of testing and greater adaptability to cope with new threats from infectious microorganisms. The combination of the new tools that are now being developed in research laboratories should lead to profound changes in the way that clinical microbiologists work.

Microorganisms as research tools

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References List PM., Dunn (1996). Dr Edward Jenner (1749-1823) of Berkeley, and vaccination against smallpox. Arch. Dis. Child. Fetal Neonatal Ed., page 74 Stanley, G. (1998). Microbiology of Fermented Foods, 2nd ed., vol. 1, page 263307 The European Parliament and the council of the European Union (2001). Directive on the release of genetically modified organisms (GMOs) Directive 2001/18/EC ANNEX I A., Official Journal of the European Communities, page 17