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    Production of Antibiotics

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    Rajesh Sharma
    The document discusses the production of several antibiotics including penicillin, streptomycin, chloramphenicol, and griseofulvin. It describes the microbes used to produce each antibiotic, such as Penicillium chrysogenum for penicillin and Streptomyces venezuelae for chloramphenicol. It also summarizes the fermentation processes, isolation and purification methods for each antibiotic. The production of penicillin involves a fed-batch fermentation with P. chrysogenum followed by precipitation to isolate the antibiotic. Streptomycin uses a three-phase fermentation with S. griseus and is recovered via adsorption and precipitation.

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    Production of Antibiotics

    Uploaded by

    Rajesh Sharma
    512 views30 pages
    The document discusses the production of several antibiotics including penicillin, streptomycin, chloramphenicol, and griseofulvin. It describes the microbes used to produce each antibiotic, such as Penicillium chrysogenum for penicillin and Streptomyces venezuelae for chloramphenicol. It also summarizes the fermentation processes, isolation and purification methods for each antibiotic. The production of penicillin involves a fed-batch fermentation with P. chrysogenum followed by precipitation to isolate the antibiotic. Streptomycin uses a three-phase fermentation with S. griseus and is recovered via adsorption and precipitation.

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    14349_Production of Antibiotics

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    The document discusses the production of several antibiotics including penicillin, streptomycin, chloramphenicol, and griseofulvin. It describes the microbes used to produce each antibiotic, such as Penicillium chrysogenum for penicillin and Streptomyces venezuelae for chloramphenicol. It also summarizes the fermentation processes, isolation and purification methods for each antibiotic. The production of penicillin involves a fed-batch fermentation with P. chrysogenum followed by precipitation to isolate the antibiotic. Streptomycin uses a three-phase fermentation with S. griseus and is recovered via adsorption and precipitation.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    512 views30 pages

    Production of Antibiotics

    Uploaded by

    Rajesh Sharma
    The document discusses the production of several antibiotics including penicillin, streptomycin, chloramphenicol, and griseofulvin. It describes the microbes used to produce each antibiotic, such as Penicillium chrysogenum for penicillin and Streptomyces venezuelae for chloramphenicol. It also summarizes the fermentation processes, isolation and purification methods for each antibiotic. The production of penicillin involves a fed-batch fermentation with P. chrysogenum followed by precipitation to isolate the antibiotic. Streptomycin uses a three-phase fermentation with S. griseus and is recovered via adsorption and precipitation.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    of 30

    Production of Penicillin

    Downstream Processing

    • Products in a fermenter are impure and dilute, so


    need to be purified by downstream processing.
    • This usually involves filtration to separate the
    microbial cells from the liquid medium, followed by
    chemical purification and concentration of the
    product
    • Downstream processing can account for 50% of
    the cost of a process.
    • Antibiotics are antimicrobial agents
    produced naturally by other microbes
    (usually fungi or bacteria).

    • The first antibiotic was discovered in


    1896 by Ernest Duchesne and
    "rediscovered" by Alexander Flemming
    in 1928 from the filamentous fungus
    Penicilium notatum.
    • The antibiotic substance, named penicillin,
    was not purified until the 1940s (by Florey
    and Chain), just in time to be used at the end
    of the second world war.
    • Penicillin was the first important commercial
    product produced by an aerobic, submerged
    fermentation
    • When penicillin was first made at the end of
    the second world war using the fungus
    Penicilium notatum, the process made 1 mg

    • Today, using a different species (P.


    chrysogenum) and a better extraction
    procedures the yield is 50 g

    • There is a constant search to improve the


    yield.
    • Antibiotics can be selectively toxic by
    targeting such features as the bacterial
    cell wall, 70S ribosomes , and enzymes
    that are specific to bacteria.

    • In this way the human eukaryotic cells


    are unaffected.
    For example:
    • penicillin, ampicillin, amoxycillin, methicillin
    • Inhibits enzymes involved in synthesis of
    peptidoglycan for bacterial cell wall, causing
    cell lysis.
    • Bacteriocidal
    • Narrow spectrum- little effect on Gram
    negative cells.
    Other antibiotics may affect:
    • Cell membrane
    • DNA replication
    • Transcription
    • Translation
    Antibiotic production
    • There are over 10 000 different antibiotics
    known, but only about 200 in commercial
    use, since most new antibiotics are no better
    than existing ones.
    • There is a constant search for new
    antibiotics. Antibiotics are the most-
    prescribed drugs and are big business.
    • Finding a new antibiotic and getting it on to
    the market is a very long process and can
    take 15 years.
    • Figure shows the kinetics of the
    penicillin fermentation with
    Penicillium chrysogenum.
    • 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.
    • 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.
    Antibiotic Production Methods
    • Antibiotics are produced on an industrial scale
    using a variety of fungi and bacteria.
    • Penicillin is produced by the fungus Penicillium
    chrysogenum which requires lactose, other sugars,
    and a source of nitrogen (in this case a yeast
    extract) in the medium to grow well.
    • Like all antibiotics, penicillin is a secondary
    metabolite, so is only produced in the stationary
    phase.
    • It requires a batch fermenter, and a fed batch
    process is normally used to prolong the stationary
    period and so increase production.
    • Downstream processing is relatively easy
    since penicillin is secreted into the medium
    (to kill other cells), so there is no need to
    break open the fungal cells.

    • However, the product needs to be very pure,


    since it being used as a therapeutic medical
    drug, so it is dissolved and then precipitated
    as a potassium salt to separate it from other
    substances in the medium.
    • The resulting penicillin (called penicillin G)
    can be chemically and enzymatically
    modified to make a variety of penicillins with
    slightly different properties.

    • These semi-synthetic penicillins include


    penicillin V, penicillin O, ampicillin and
    amoxycillin.
    PRODUCTION OF STREOPTMYCIN
    Streptomycin and various other antibiotics are produced
    using strains of Streptomyces griseus. Spores of this
    actinomycete are inoculated into a medium to establish a
    culture with a high mycelial biomass for introduction into an
    inoculum tank, with subsequent use of the mycelial inoculum
    to initiate the fermentation process in the production tank. 
    The medium contains soybean meal (N- source), glucose (C-
    source) and NaCl.
    Temperature: 28°C
    pH: 7.6-8.0.
    High agitation and aeration are needed.
    Fermentation Period: 10 days.
    The classic fermentation process involves three phases.

    During the first phase there is rapid growth of the microbe with
    production of mycelial biomass. 
    Proteolytic activity of the microbe releases NH3 to the
    medium from the soybean meal, causing a rise in pH.
    During this initial fermentation phase there is little
    production of streptomycin. During second phase there is
    little additional production of mycelium, but the secondary
    metabolite, streptomycin accumulates in the medium. The
    glucose and NH3 released are consumed during this
    phase.
    The pH remains fairly constant-between 7.6 and 8.0. In the
    third and final phase, when carbohydrates become depleted,
    streptomycin production ceases and the microbial cells
    begin to lyse pH increases and process normally ends by
    this time. 

    After the process is complete, mycelium is separated from


    the broth by filtration and the antibiotic recovered. In one
    method of recovery and purification, streptomycin is
    adsorbed onto activated charcol and eluted with acid
    alcohol. It is then precipitated with acetone and further
    purified by use of column chromatography
    PRODUCTION OF CHLOREMPHENICOL
    Isolated from Streptomyces venezuelae in 1947

    Inhibitory activity against both gram positive and gram


    negative bacteria

    Also effective against actinomycetes, mycobacteria and


    anaerobic bacteria

    Quite effective but having severe side effects

    It affects bone marrow 


    Submerged Aerobic Fermentation

    The medium contains

    Carbon source: Glycerol, molasses and dried distillers grain


    slops

    Nitrogen source: Yeast fermentation solubles, hog residues,


    tryptone, etc.

    Mineral Nutrients: Sodium chloride, zinc and iron in traces

    pH: 7.5

    Temperature: 25 degree centigrade

    Period of Fermentation: 10-15 days


    Isolation and Purification:

    Extracted from clarified broth

    Extracts are concentrated, diluted with kerosene and


    washed with diluted acids, alkalis and water.

    Lipids with petroleum ether are removed

    Passed through column of charcoal or alumina

    The product is then crystallized


    PRODUCTION OF GRISEOFULVIN
    Isolated from Penicillium patulum, P. raistrickii, P. urticase

    Inhibitory activity against fungi

    Treatment and control of fungal diseases of skin, hair, nails


    caused by the dermatophytes.

    Strain with conidia yields maximum products

    It is less toxic and systemic 


    Submerged Aerobic Fed Batch Fermentation

    The medium contains

    Carbon source: Malted cereal extracts, glucose, soluble starch

    Nitrogen source: Whey powder nitrogen, protopeptone, corn


    steep liquor

    Mineral Nutrients: Calcium carbonate, Potassium chloride,


    Potassium Dihydrogen phosphate

    pH: 6.8-7.2

    Temperature: 25 degree centigrade

    Period of Fermentation: 10 days


    Phosphate and chlorine are required in sufficient amount
    otherwise the obtained product is devoid of antifungal
    property.

    Carbohydrate glucose syrup (50%) is added stepwise.

    Air flow is required

    Yield: 6-8g/Litre

    To increase the yield methyl donors may be added to the


    medium
    Extraction and Purification:

    Treated with organic solvents and removed by filtration

    Wet mycelia treated with butyl acetate followed by


    subsequent evaporation

    Crude griseofulvin is washed with chloroform and crystallized

    For higher production extraction from mycelium is done with


    methylene chloride and recrystallization with acetone.

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