This document describes a student project on the production of the antibiotic erythromycin. It provides background on erythromycin, including its discovery in 1952. It then discusses the industrial production process, which involves fermentation of the bacteria Streptomyces erythreus to produce erythromycin. Key steps include inoculum development, fermentation, isolation and extraction of erythromycin from the fermentation broth. The document also summarizes methods explored by the students to improve the erythromycin production strain and discusses clinical uses and side effects of erythromycin.

1. PROJECT TITLE
PRODUCTION OF ERYTHROMYCIN
GROUP MEMBERS
UMESH VERMA 17BTBIOCE003
HARSHITA RAI 17BTBIOCE004
TUSHAR MISHRA 17BTBIOCE005
ATAUL ADNANI 17BTBIOCE009
NITINJAY SINGH 17BTBIOCE011

2. CONTENT
1. INTRODUCTION
HISTORY OF ERYTHROMYCIN
GENRAL CHARACTERISTIC ( UMESH VERMA )
2. MECHANISM OF ACTION ,PHARMACOKINETICS
BIOSYNTHESIS OF ERYTHROMYCIN ( HARSHITA RAI )
3. INDUSTRIAL PRODUCTION ( TUSHAR MISHRA )
INNOCULUM DEVELOPMENT, FERMENTATION
4. ISOLATION & EXTRACTION ( ATAUL ADNANI )
5. IMPROVEMENT OF STRAIN & CLINICAL USES
( NITINJAY SINGH )

3. INTRODUCTION
 Erythromycin belongs to the group of Macrolide antibiotics.
 Macrolide antibiotics slow the growth of, or sometimes kill
sensitive bacteria by reducing the production of important
proteins needed by the bacteria to survive.
 It has an antimicrobial spectrum similar to or slightly wider
than that of penicillin and is often used for people who have
allergy to penicillin.

4. CONTINUED…
 Erythromycin is produced using Streptomyces.
 Streptomyces is a genus of Gram-positive bacteria that grows in
various environments, with a filamentous form similar to fungi.
 The commonly used macrolides are:
 Erythromycin.
 Clarithromycin.
 Roxithromycin
 Azithromycin

5. HISTORY
 Erythromycin was discovered in 1952 .
 Eli Lilly’s research team, led by J. M. McGuire managed
to isolate erythromycin from the metabolic products of a
strain of Streptomyces erythreus.
 Found in a soil sample from the Philippines.

6. GENERAL CHARACTERISTICS
 Erythromycin contains three characteristics parts in the molecule:
 A highly substituted macrocyclic lactone: aglycone
 It is a macrocyclic compound containing 14-membered lactone ring
with 10 asymmetric centers.
 A ketone group.
 An amino deoxysugar: glycon
 Its chemical formula is C37 H67NO13
 Molecular Weight 733.937 g/mol

7. Physical and Chemical properties
 It is a crystalline, colorless compound .
 Slightly soluble in water but dissolves easily in most of the common organic
solvents.
 Crystals are obtained readily from aqueous acetone, aqueous alcohol or
chloroform.
 Odorless
 Taste is Bitter

8. MECHANISM OF ACTION
 Erythromycin displays a bacteriocidal activity particularly at
higher concentrations.
 It prevents the growth of bacteria by inhibiting their protein
synthesis.
 Erythromycin binds to the 23s rRNA molecule in the 50S
ribosomal subunit.
 Binding site near the peptidyl transferase center.
 This binding blocks the exit of the growing peptide thus inhibiting
the translocation of the peptides.

9. MECHANISM

10. Pharmacokinetics
 Erythromycin is easily inactivated by gastric acids, therefore all
orally administered formulations are given as either enteric
coated or more stable salts or esters, such as erythromycin
ethyl succinate.
 It is rapidly absorbed and diffused into most tissues and
phagocytes.
 Due to high concentration in phagocytes, erythromycin is
actively transported to the site of infection, where during active
phagocytosis, large concentration of erythromycin is released

11. Erythromycin Biosynthesis.
 The three genes eryAI–III encode for the multidomain proteins DEBS1–3.
 The first protein, DEBS1, consist of a loading module with an
actyltransferase (AT) and acyl carrier protein (ACP) domain, followed by
module 1 with the ketosynthase (KS), AT, ketoreductase (KR), and an ACP
domain, followed by module 2 with KS, AT, KR, and ACP domains.
 The second protein, DEBS2, consists of two modules, with a KS, AT, ACP
domain and a KS, AT, dehydrogenase (DH), enoylreductase (ER), KR, ACP
domain, respectively.
 The third protein, DEBS3, also consists of two modules and a thioesterase
(TE) domain to release the polyketide from the module.

12. INDUSTRIAL PRODUCTION
 Erythromycin is produced from Streptomyces and
Micromonospora.
 Erythromycin is produced mainly by submerged
fermentation.
 During fermentation, different types of erythromycin
are produced like erythromycin A, B, C, and D.
 The steps for erythromycin production are as follows:

13. Inoculum Development
 Inoculum is prepared from suspension of at least 108 cells/ml.
 Sporulation of S.erythraeus is done on tryptone agar slant.
 Cells are harvested from agar plates and suspension is taken in sterile water and
stored at 4ºC .
2. Medium:
 Sucrose/ starch
 Corn steep liquor
 Soyabean oil meal
 Yeast
 NaCl
 CaCo3 precipitate
 pH
 Water

14. S. erythraea Fermentation
 Fermentation is carried out in stirred tank fermenter.
 The batch starts with sterilization followed by pumping
deionized water into the fermenter.
 The organism is grown in the broth for 4 days at a
temperature of 26° C

15. Cont..
 Addition of n-propanol increases the production of erythromycin.
 Samples were drawn aseptically through a cross flow filter assembly .
 Production of Erythromycin production occurs when froth reaches
stationary phase.
 Daily samples of 50 or 100 mL were drawn to determine the
erythromycin concentration.
 Samples were stored at -20 °C.

16. Isolation & Extraction of Erythromycin
 Mycelium is separated from the broth by filter press,
centrifugation or drum filtration.
 Acidic condition helps to separate mycelium the broth.
 Acidic condition is maintained by addition of Butyl acetate which
favors the separation of the mycelium. Also it dissolves the
antibiotic.
 It is then washed with water.

17. Extraction
 It is extracted using methyl iso butyl ketone or ethyl acetate.
 It is then transferred to acidic water.
 pH is adjusted with HCl, acetic acid and citric acid.
 Purification and concentration is carried out in ion exchange
resin amberlite 50.
 Elution is carried out by a mixture of organic solvents and water
at pH 3 to 8.
 Erythromycin is obtained as dihydrate salt.
 The dehydrate crystals of erythromycin are filtered and dried on
vacum tray dryer..

18. IMPROVEMENT OF STRAIN
 Saccharopolyspora erythraea MTCC 1103 is used for the enhanced
production of erythromycin.
 Strain improvement was done by random UV-mutagenesis.
 Mutant strain showed 40 % higher yield in production medium as
compared to wild strain.
 Erythromycin potency assay and HPLC analysis were performed to confirm
the presence of erythromycin in the partially purified samples.
 Effects of various parameters such as bagasse concentration, organic
nitrogen source, inorganic nitrogen source, pH and temperature.

19. CARBON SOURCE
 Bagasse can be used as an alternate carbon source in erythromycin
medium.
 Erythromycin production medium was found to be 512 mg/L which was 28 %
higher than glucose based medium.
NITROGEN SOURCE
 It was found that corn steep liquor was best as compared to yeast extract,
malt extract, casein and peptone.
 For corn steep liquor, concentration of erythromycin 416 mg/L.
 Yeast extract and malt extract 356 and 310 mg/L
 Peptone added medium 120 mg/L

20. ERYTHROMYCIN CONCENTRATION

21. CONT…
pH
 Erythromycin production rate was high at pH 7
 At pH 7, the concentration of erythromycin was found to be 461 mg/L
and dry weight of cell biomass as 500 mg/L.
Temperature
 The optimum temperature for erythromycin production was found to be 28
°C

22. CLINICAL USES
 Erythromycin is an antibiotic useful for the treatment of a number of
bacterial infections.
 This includes respiratory tract infections, skin infections, chlamydia infections,
pelvic inflammatory disease, and syphilis.
 Erythromycin may be used to improve delayed stomach emptying.
 It can be given intravenously and by mouth. An eye ointment is routinely
recommended after delivery to prevent eye infections in the newborn.

23. CONT…
 Penicillin substitute in penicillin allergic individuals
 In respiratory, neonatal, ocular, or genital chlamydial infections.
 Treatment of community-acquired pneumonia.
 Used in covid 19 with hydroxychloroquine.
 It may also be used during pregnancy to prevent Group B streptococcal
infection in the newborn.
 Prophylaxis against endocarditis during dental procedures in individuals with
valvular heart disease.

24. SIDE EFFECTS OF ERYTHROMYCIN
 Inflammation of the liver.
 Confusion or hallucinations.
 Kidney inflammation or infection.
 Abdominal pain.
 vomiting

25. THANK YOU