Tetracyclin and cephalosporins are one of the major used antibiotics commonly all around the world. They are used to treat against microorganisms as a bactericidal, these eliminates those organisms in the host through various mechanism. These antibiotics are produced in a large scale using a bioreactors in many countries.

2. Tetracycline, Lloyd H. ConoverLloyd H. Conover is credited with having invented the
first antibiotic made by chemically modifying a naturally-produced
drug. His creation, Tetracycline, has been one of the most
frequently prescribed antibiotics in the United States for treating
bacterial infections for several decades.

3. • Tetracyclines are a class of antibiotics, which are chemical
substance produced by a microorganism that are able to kill other
microorganism without being toxic to the person, animal (or )plant.
• Tetracyclines are derived directly from a bacterium known as
Streptomyces coelicolor / Streptomyces aureofaciens. .

5. • Tetracycline inhibits protein synthesis by blocking the attachment of
charged aminoacyl-tRNA to the A site on the ribosome. Tetracycline
binds to the 30S subunit of microbial ribosomes. Thus, it prevents
introduction of new amino acids to the nascent peptide chain.
• It exerts a bacteriostatic effect on bacteria by binding reversible to
the bacterial 30S ribosomal subunit and blocking incoming
aminoacyl tRNA from binding to the ribosome acceptor site.

7. • The Tetracyclines was produced through Shaker Flask
Fermentations of Streptomyces coelicolor / Streptomyces
aureofaciens.
• Production of chlortetracycline, all media contained less than 1 ppm
chloride. The base synthetic medium which developed for th
fermentation and has the following formulation (per L):
– Sucrose, 30 g; Trisodium citrate, 2.5 g; Ammonium sulfate, 3.3 g;
– Magnesium sulfate heptahydrate, 0.25 g; Monopotassium phosphate,0.10g;
– Dipotassium phosphate, 0.10 g; Calcium carbonate, 1.00 g;
– Manganese sulfate, 0.01 g; Zinc sulfate, 0.04 g;
– Potassium dichromate, 0.016 mg; and Acetic acid, 0.40 ml.

8. • Isolates are carried on Waksman agar for the isolation of soil
microorganism (Waksman, 1922) without adjustment of pH, and are
incubated at 30 C for a period of 10 days to permit good
sporulation. The formula for this agar follows (per L):
– Glucose, 10.0 g;
– peptone, 5.0 g;
– Monopotassium phosphate, 1.0 g;
– Magnesium sulfate heptahydrate, 0.5 g;
– Agar, 20.0 g.
Selected isolates are stored under oil or lyophilized.

9. • Medium (per L):
– Sucrose, 30.0 g;
– Soybean meal, 5.0 g;
– Sodium Citrate, 1.0 g;
– Ammonium sulfate, 3.3 g;
– Magnesium sulfate heptahydrate, 0.25 g;
– Monopotassium phosphate, 0.10 g;
– Dipotassium phosphate, 0.10 g;
– Calcium carbonate, 1.00 g;
– Manganese sulfate, 0.01 g;
– Zinc sulfate, 0.04 g;
– Potassium dichromate, 0.016 mg;
– Acetic acid, 0.40 ml.

10. • Volume: 50 ml (culture) per 250-ml (medium); 400 ml per 2000-ml.
• Shaker: stroke at 97 cycles per min.
• Temperature: 30° C.
• Inoculum: dry mycelial-spore transfer, liquid mycelial suspension, or
lyophile.
• Period of incubation: 48 to 72 hr to give pH ranging between 4.5
and 5.0.

11. • Medium (per L):
– Sodium Citrate, 12.8 g;
– Sucrose, 40.0 g;
– Ammonium sulfate, 6.0 g;
– Magnesium sulfate heptahydrate, 0.25 g;
– Monopotassium phosphate, 0.15 g;
– Calcium carbonate, 11.00 g;
– Manganese sulfate, 0.01 g;
– Zinc sulfate, 0.04 g;
– Potassium dichromate, 0.016 mg.

12. • Volume: 50 ml per 250-ml Erlenmeyer flask.
• Shaker: rotary at 200 rpm.
• Temperature: 30 C.
• Inoculum: 5 per cent mycelial transfer.
• Period of incubation: 4 to 6 days.
• Preparation of sample for assay: Whole broth cultures
are acidified to pH 2.0 to 2.5 with 5 N sulfuric acid and
held for 1hr before filtration through Whatman No. 4
paper. The paper disc method of assay with E.coli
strain ATCC no. 9637 is employed. Assays are run
against a tetracycline standard.

14. • At the end of the fermentation, the culture broth is filtered to
remove the mycelium.
• The filtrate is treated with n-butanol or methylisobutylketone in
acidic or alkaline condition for extracting the antibiotic.
• It is then absorbed to activated charcoal to remove other impurities.
Tetracycline is eluted and crystallized.

16. • The cephalosporins are a class of β-lactam antibiotics originally
derived from the fungus Acremonium, which was previously known
as "Cephalosporium". Together with cephamycins, they constitute a
subgroup of β-lactam antibiotics called cephems. Cephalosporins
were discovered in 1945, and first sold in 1964.
STRUCTURE OF CEPHALOSPORIN

17. • The aerobic mould (Cephalosporium acremonium, later renamed as
Acremonium chreysogenum) which yielded cephalosporin C was found in
the sea near a sewage outfall of Cagliari harbour in Sardinia, by
the Italian pharmacologist Giuseppe in July 1945.
• Cephalosporins are indicated for the prophylaxis and treatment of
infections caused by bacteria.
• First-generation cephalosporins are active predominantly against Gram-
positive bacteria, they are therefore used mostly for skin and soft tissue
infections.
• Successive generations of cephalosporins have increased activity
against Gram-negative bacteria, albeit often with reduced activity against
Gram-positive organisms.
• The antibiotic may be used for patients who are allergic to penicillin. The
different β-lactam antibiotic structure. The drug is able to be excreted in
the urine.

18. • Cephalosporins are bactericidal and have the same mode of action
as other β-lactam antibiotics (such as penicillins), but are less
susceptible to β-lactamases. Cephalosporins disrupt the synthesis of
the peptidoglycan layer forming the bacterial cell wall. The
peptidoglycan layer is important for cell wall structural integrity.

19. • The fermentation process concerned with the production of
cephalosporin is similar to that of penicillin.
• The culture media consists of corn steep liquor and soy flour-based
media in a continuous feeding system.
• The other ingredients of the medium include sucrose, glucose and
ammonium salts.
• Methionine is added as a source of sulfur.

20. • The fermentation is carried out at temperature 25-28°C and pH 6-7.
The growth of micro-organisms substantially increases with good
O2supply, although during production phase, O2consumption
declines.
• By using Penicillin V as the starting material, chemical synthesis of
cephalosporin has become possible. This is being done due to low
cost of production of penicillin.

21. • Cephalosporin C from the culture broth can be recovered by ion-
exchange resins, and by using column chromatography.
Cephalosporin C can be precipitated as zinc, sodium or potassium
salt, and isolated.