Priyo Wahyudi

Lecture 15Lecture 15PRODUK MIKROBIOLOGI DI BIDANG PRODUK MIKROBIOLOGI DI BIDANG

FARMASIFARMASI

FARMASI – FMIPA – UHAMKA2010

Produk mikrobiologi di bidang farmasi

a. Makanan dan minumanb. Vaksin konvensionalc. Molekul berukuran kecil (asam amino, vitamin,

antibiotik)

Makanan dan Minuman

4

Contoh Produk Fermentasi :

Makanan dan Minuman Fermentasi

• Tempe fermentasi kedelai oleh Rhizopus sp• Oncom fermentasi limbah kulit kacang/kedelai oleh

Monilia sitophyla• Tape fermentasi singkong oleh Saccharomyces

cerevisiae• Keju fermentasi susu • Nata fermentasi gula oleh Acetobacter xylinum• Sosis fermentasi daging oleh Lactobacillus brevis• Sauerkraut fermentasi sayuran oleh Lactobacillus

plantarum

Makanan dan Minuman Fermentasi

• Kecap fermentasi kedelai oleh Aspergillus wentii• Yoghurt susu fermentasi oleh Bakteri Asam Laktat• Bir fermentasi malt (starch) menjadi alkohol oleh

Saccharomyces cerevisiae• Wine fermentasi buah menjadi alkohol oleh

Saccharomycer cerevisiae• Asam cuka fermentasi gula oleh Acetobacter aceti• Biomassa Saccharomyces cerevisiae

Sugar Ethanol + CO2

Saccharomyces cerevisiae

Malic acid Lactic acidLactic acid bacteria

Ethanol Acetic acidAcetobacter or Gluconobacter

Microbial Metabolism

Fermentation Technology

Making Red Wine

Vinegar Production

• The active ingredient in vinegar is acetic acid, which is produced by acetic acid bacteria oxidizing an alcohol-containing fruit juice (Figure 30.22).

• Adequate aeration is the most important consideration in ensuring a successful vinegar process.

Diagram of a vinegar generator.

Yeast as a Food and Food Supplement

• Yeast cells are grown for use in the baking and food industries

Vaksin Konvensional

What is a Vaccine?

• A vaccine is a non-pathogenic antigen that mimics a particular pathogen in order to elicit an immune response as if that actual pathogen were in the body.

• The overall goal of a vaccine is to establish immunity against that particular pathogen.

Types of Vaccines

• There are numerous types of vaccines.• Each different type of has its own unique

properties.• There function, however, is the same, to

establish immunity against a particular pathogen.

1. Attenuated Virus/Bacteria

• These vaccines consist of live, but weakened, viruses or bacteria.

• These organisms have been altered, either genetically or chemically, in a way that they are not pathogenic.

• An example is the attenuated virus vaccine for yellow fever, which utilizes the YF17D strain, a weakened form of the wild virus.

2. Killed Whole Organism

• This vaccine consist of the actual pathogen, however, it has been killed, either by a heat treatment or chemically.

• An example is the Salk vaccine for polio, which utilizes whole polioviruses that have been inactivated by formaldehyde.

3. Toxoids

• Some species of bacterial produce what is known as exotoxins.

• Toxoids are vaccines which consist of exotoxins that have been inactivated, either by heat or chemicals.

• These vaccines are intended to build an immunity against the toxins, but not necessarily the bacteria that produce the toxins.

• Some examples are botulinum antitoxen and diphtheria antitoxen.

4. Surface Molecules

• Proteins, carbohydrates, and lipids, that are found on the surface of pathogens, are isolated and used as a vaccine.

• Proteins are usually large and complex enough to be used on there own.

• Carbohydrates and lipids requires conjugated with a large protein in order to be immunogenic.

• An example of surface molecules used as a vaccine are hepatitis B surface antigens.

5. Anti-Idiotype Vaccines

• In this unique type of vaccine, antibodies from a sick individual are isolated.

• These antibodies are then injected into a lab animal, which may then produce an antibody whose antigen binding site mimics the epitope that the original antibody binds to.

• These antibodies are then isolated and injected into a healthy individual, who may produce antibodies with an antigen binding site that binds to the antigen binding site of the animals antibodies.

5. Anti-Idiotype Vaccines

• Because the animals binding site resembles the epitope of an antigen on a particular pathogen, the individual will have an immunity against that pathogen.

6. DNA Vaccines

• DNA vaccines consist of plasmids that contains genes for certain types of antigens.

• Once administered, the plasmid is taken up by the target cell and the genes are expressed.

• The cell then either excretes the antigen or displays it on an MHC-I molecule.

7. Chimeric Vaccines

• Chimeric vaccines usually consist of attenuated viruses that have been engineered to carry antigens from multiple types of pathogens.

• For example, the yellow fever vaccine YF17D has been engineered to carry antigens from HIV, different types of bacteria, malaria, even cancer.

• The main of a chimeric vaccine is the establishment of immunity against several different diseases with one administration.

Vaccine Production Methods

• There are three main vaccine manufacturing strategies:– In-vivo– In-vitro– Chemical Synthesis

• Some vaccines can be produced using any one of the three methods while for other vaccines, only one method will work.

In-Vivo

• In in-vivo manufacturing, the vaccine is produced inside a living organism.

• Embryonated Chicken eggs are are commonly used, particularly in producing flu vaccines.

• Vaccines, such as anti-idiotype, can also be produced in lab animals, such as mice.

• There are even some species of plant, such as bananas, that have been genetically engineered to produce a vaccine.

In-Vitro

• Here, using recombinant DNA technology, vaccines can be produced in yeast cultures, bacterial cultures, or cell cultures.

• Recombinant vaccines, such as chimeric vaccines, are produced in this manor.

• Attenuated virus/bacteria vaccines can also be produced this way.

Chemical Synthesis

• Here, instead of using biological systems to produce a vaccine, a vaccine can be produced in a lab.

• Vaccines that utilize synthetic peptides as well as conjugated lipids and polysaccharides are manufactured this way.

• Usually, this method is used in combination with either in-vivo or in-vitro production.

Asam amino, Vitamin & Antibiotik

Vitamins and Amino Acids

Vitamins produced microbially include vitamin B12 and riboflavin

Amino Acid

• The most important amino acids produced commercially are glutamic acid, aspartic acid, phenylalanine, and lysine

• Aspartic acid and phenylalanine are the ingredients of the artificial sweetener aspartame, a non-nutritive sweetener of diet soft drinks and other foods sold as low-calorie or sugar-free products.

• High yields of amino acids are obtained by modifying regulatory signals that control synthesis of the particular amino acid such that overproduction occurs.

Lists amino acids used in the food industry

Steroids and the Biotransformation Process

Microbial biotransformation employs microorganisms to biocatalyze a specific step or steps in an otherwise strictly chemical synthesis (Figure 30.14).

Enzymes as Industrial Products• Microorganisms are ideal for the large-scale production

of enzymes. Many enzymes are used in the laundry industry to remove stains from clothing, and thermostable and alkalistable enzymes have many advantages in these markets.

• Certain enzymes are produced in large amounts by some organisms, and instead of being held within the cell, they are excreted into the medium.

• Extracellular enzymes, called exoenzymes, can digest insoluble polymers such as cellulose, protein, and starch. The products are then transported into the cell where they are used as nutrients for growth.

• The term extremozyme has been coined to describe enzymes that function at some environmental extreme, such as high temperatures or low pH (Figure 30.15). Enzymes from extremophiles are desirable for biocatalyses under extreme conditions.

• For some biocatalytic processes, it is desirable to fix soluble enzymes onto a solid surface. These are called immobilized enzymes (Figure 30.16).

Table 30.4 lists microbial enzymes and their applications.

Citric Acid & Organic Compounds

• A number of organic chemicals are produced commercially by use of microorganisms, of which the most important economically is citric acid, produced by Aspergillus niger

Fermentation Technology

Fermentation Technology

Production Curve of a Primary Metabolite

Figure 28.11b

Production Curve of Secondary Metabolite

Yeast Fermentations

The Birth of Modern Chemotherapy

• 1928: Alexander Fleming discovered the first antibiotic.

• He observed Penicillium fungus made an antibiotic, penicillin, that killed S. aureus.

• 1940s: Penicillin was tested clinically and mass produced.

Interrelationship of the main primary metabolic pathway for aromatic amino acid synthesis, and the secondary metabolic pathways for a variety of antibiotics.

• Many economically valuable microbial products are secondary metabolites.

• Large-scale industrial fermentation presents several engineering problems. Aerobic processes require mechanisms for stirring and aeration

• The microbial process must be continuously monitored to ensure satisfactory yields of the desired product.

Fermentor sizes for various industrial processes

Fermentation Scale-Up

Fermentation scale-up is the process of gradually converting a useful industrial fermentation from laboratory scale to production scale. Aeration is a particularly critical aspect to monitor during scale-up studies.

A brief history of antibiotics• 1495, mercury to treat syphilis.• 1630, quinine (chinchona tree) for malarial fever by South American

Indians.• 1889, Buillemin defined antibiosis.• 1910, Paul Ehrlich developed arsenical compound (Salvarsan) for

syphilis, term: the chemical knife.• 1929, Alexander Fleming found penicillin.• 1935, Gerhard Domagk showed the value of sulfonamides.• 1940, Ernst Chain and Howard Flory demonstrated the effect of

penicillin.• 1940-1970, then searching for new antibiotics• ~ recent year: modifying old drugs, finding new discipline in

antibacterial combats• Early time in war: thanks penicillin, we can go home now

Thanks to work by Alexander Fleming (1881-1955), Howard Florey ( 1898-1968) and Ernst Chain (1906-1979), penicillin was first produced on a large scale for human use in 1943. At this time, the development of a pill that could reliably kill bacteria was a remarkable development and many lives were saved during World War II because this medication was available.

E. Chain H. FloreyA. Fleming

A tale by A. Fleming

He took a sample of the mold from the contaminated plate. He found that it was from the penicillium family, later specified as Penicillium notatum. Fleming presented his findings in 1929, but they raised little interest. He published a report on penicillin and its potential uses in the British Journal of Experimental Pathology.

Fleming and Penicillin

Microbial Sources of Antibiotics

The industrial production of antibiotics begins with screening for antibiotic producers

Once new producers are identified, purification and chemical analyses of the antimicrobial agent are performed.

If the new antibiotic is biologically active in vivo, the industrial microbiologist may genetically modify the producing strain to increase yields to levels acceptable for commercial development.

Industrial Production of Penicillins and Tetracyclines

Major antibiotics of clinical significance include the -lactam antibiotics penicillin (Figure 30.9) and

cephalosporin and the tetracyclines (Figure 30.11).

• Cephalosporins are valued clinically not only because of their low toxicity but also because they are broad-spectrum antibiotics, useful against a wide variety of bacterial pathogens.

• If the penicillin fermentation is carried out without addition of side-chain precursors, the natural penicillins are produced. The fermentation can be more directed by adding to the broth a side-chain precursor so that only one desired penicillin is produced.

Figure 30.10 shows the kinetics of the penicillin fermentation with Penicillium chrysogenum.

• The product formed under these conditions is referred to as a biosynthetic penicillin. To produce the most useful penicillins, those with activity against Gram-negative Bacteria, a combined fermentation and chemical approach is used that leads to the production of semisynthetic penicillins.

• All of these antibiotics are typical secondary metabolites, and their industrial production is well worked out despite the fact that the biochemistry and genetics of their biosynthesis are only partially understood.

Thank You