Tech Seminar
Tech Seminar
ROLL BOTTLE METHOD: Large surface area for cultivation of spores 300c.c of medium with 3% agar is sterilized in 1dm3 bottles. Cooled to 45deg.C
On rotation in roller mill, agar sets as a cylindrical shell inside the bottle
Inoculation with a spore suspension of P.chrysogenum and incubation at 24 for 6 to 7 days. Roux bottle is a similar method used in production of clavulanic acid from S.clavuligerus
Organism
Media contents
Glycerol Cane molasses Curbay BG MgS04 7H2O KH2P04 Peptone NaCl Agar Molasses KH2P04 Agar
Amount added(g/dm3)
7.5 7.5 2.5 0.05 0.06 5.0 4.0 20 300 0.5 20
P.chrysogenum
Substrates used are barley, hard wheat bran, ground maize and rice. Filamentous organisms will sporulate profusely on the surface of cereal grains
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FACTORS:
The amount of water added to the cereal before sterilization Relative humidity of the atmosphere, which should be high
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Organism
Substrate
200 grams of 'pot barley 100grams of moistened wheat bran
Conditions,yield
5 X 10^11 conidia After 6 days at 28 Deg.C and 98% RH
Aspergillus ochraces
Penicillium , cephalosporium
Aspergillus, Penicillium S.aureofaciens
Cooked rice
White bread Millet
Organism used: P.patulum For prolific sporulation the nitrogen level must be limited to between 0.05 and 0.1% wIv and good aeration must be maintained The lower the degree of aeration, the lower the concentration of nitrogen needed to induce sporulation. Incubation at 25 for 7 days Yield : 10% inoculum for a vegetative seed stage in a stirred fermentor
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4. 5.
Fermentation spore inoculum - penultimate stage:penicillium production - early stages : sagamycin - Reduces cost of installation and operation of seed tanks vegetative inoculum - eg: clavulanic acid pdtn - Reduces fermentation time - High labour cost
Choice of inoculum depends on: Length of fermentation process, plant size, cost of labour.
Gibberellin production using Gibberella fujikuroi: - Long slants of potato dextrose agar (1 week at 24 deg.) - Growth scraped off and transferred to a liquid medium: 2% glucose,0.3% MgS04 7H2O,0.3% NH4CI and 0.3% KH2P04. - The medium was aerated(75 hours at 28) - Transfer to a seed fermenter Difficult to get uniform, standard inoculum Mycelium fragmented in homogeniser prior to use
The effect of the inoculum on the morphology of filamentous organisms in submerged culture
Two main factors that influence the morphology: Concentration of spores in a spore inoculum High spore inoculum filamentous growth Low conc. of spores - pellet formation Inoculum development medium Rich, complex media - varied dispersed growth Chemically defined media pelleted growth
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Hyphal form
Pelleted form
Broth
Difficulty in aeration
Eg: penicillin production by Eg: citric-acid production by Aspergillus niger P. chrysogenum Fusarium gramineatium pdtn requires hyphal length of 400 m.
Actinomycetes
Mycelial form : streptomycin by S. griseus Pelleted form : glucose isomerase pdtn by S. nigrificans Inoculum development for cephamycin C pdtn : Key factor : concentration of iron in the seed medium Pellet formation was observed to be detrimental to product formation The principles applied to the optimization of fungal inoculum development regimes are also relevant to actinomycete processes.