Steps involved in Fermentation
Fermentation process involves various steps
1. Production of an active pure high yielding Culture insufficient quantity to
inoculate in to the fermenter
2. Formulation of media used for the culturing of organism during the
development of inoculum and in production in fermenter
3. Designing of fermenter
4. Sterilization of media fermenter and auxiliary equipments
5. Growth of organism in the production fermenter under optimum conditions for
product formation
6. Extraction of product and its purification (downstream processing)
7. Disposal of the effluents produced by fermentation process
I. Production of active high yielding pure culture insufficient quantity to inoculate
in to the fermenter
Screening
It is a highly effective procedure to allow the detection and isolation of
microorganisms of interest from a large microbial population
Screening is done in several steps discriminating unwanted microorganisms and
allowing only useful microorganisms to grow.
1. Primary Screening
Primary screening includes several techniques
, i. Using pH indicators
Microorganisms producing acids or amines are usually detected by the incorporation
of pH indicators like bromothymol blue neutral red etc. into the media. The
production of these compounds is indicated by a colour change of these indicators
added in the media. For the detection of organic acids calcium carbonate is
incorporated into the media. The process of acid formation is confirmed by clear
zone but it is a fool proof method because even the presence of inorganic acids has
the potential to form clear zones.
For example if Ammonium Sulphate is incorporated instead of other organic nitrogen
source like peptone amino acid etc. the sulphate ions left after utilisation of
Ammonium ions results in acid production. Further screening is required for
detecting what type of organic acid is produced by chromatography. Colonies of
microorganism detected through primary screening which have the fermentation
potential should be purified and subculture on to slants for stock culture.
ii. Crowded plate technique
Screening can be used for searching microorganisms capable of producing useful
antibiotics this is done through crowded plate technique. This technique is used for
detecting the production of antibiotics and not for antibiotic sensitivity. IN crowded
plate technique the soil sample is serially diluted and diluted sample is plated and
incubated for the development of colonies. Antibiotic producing colonies are detected
by the presence of clear zone. The colony detected is isolated and streaked for
getting pure colonies and stocked. Primary screening technique selects not only
antibiotic producing colonies. Clear zones are also due to change in the pH value of
the media produced by the metabolism of the growing microbial colonies. Hence
further technique is required by secondary screening. Crowded plate technique has
got a limited application the antibiotic activity against specific microorganism has to
be detected further hence antibiotics screening is improved by the incorporation of
test organism as indicator this method is done as follows:
Serial dilution of the sample followed by plating and incubation of the plate till the
colonies are appeared and reached at appropriate size.
Fermentation process involves various steps
1. Production of an active pure high yielding Culture insufficient quantity to
inoculate in to the fermenter
2. Formulation of media used for the culturing of organism during the
development of inoculum and in production in fermenter
3. Designing of fermenter
4. Sterilization of media fermenter and auxiliary equipments
5. Growth of organism in the production fermenter under optimum conditions for
product formation
6. Extraction of product and its purification (downstream processing)
7. Disposal of the effluents produced by fermentation process
I. Production of active high yielding pure culture insufficient quantity to inoculate
in to the fermenter
Screening
It is a highly effective procedure to allow the detection and isolation of
microorganisms of interest from a large microbial population
Screening is done in several steps discriminating unwanted microorganisms and
allowing only useful microorganisms to grow.
1. Primary Screening
Primary screening includes several techniques
, i. Using pH indicators
Microorganisms producing acids or amines are usually detected by the incorporation
of pH indicators like bromothymol blue neutral red etc. into the media. The
production of these compounds is indicated by a colour change of these indicators
added in the media. For the detection of organic acids calcium carbonate is
incorporated into the media. The process of acid formation is confirmed by clear
zone but it is a fool proof method because even the presence of inorganic acids has
the potential to form clear zones.
For example if Ammonium Sulphate is incorporated instead of other organic nitrogen
source like peptone amino acid etc. the sulphate ions left after utilisation of
Ammonium ions results in acid production. Further screening is required for
detecting what type of organic acid is produced by chromatography. Colonies of
microorganism detected through primary screening which have the fermentation
potential should be purified and subculture on to slants for stock culture.
ii. Crowded plate technique
Screening can be used for searching microorganisms capable of producing useful
antibiotics this is done through crowded plate technique. This technique is used for
detecting the production of antibiotics and not for antibiotic sensitivity. IN crowded
plate technique the soil sample is serially diluted and diluted sample is plated and
incubated for the development of colonies. Antibiotic producing colonies are detected
by the presence of clear zone. The colony detected is isolated and streaked for
getting pure colonies and stocked. Primary screening technique selects not only
antibiotic producing colonies. Clear zones are also due to change in the pH value of
the media produced by the metabolism of the growing microbial colonies. Hence
further technique is required by secondary screening. Crowded plate technique has
got a limited application the antibiotic activity against specific microorganism has to
be detected further hence antibiotics screening is improved by the incorporation of
test organism as indicator this method is done as follows:
Serial dilution of the sample followed by plating and incubation of the plate till the
colonies are appeared and reached at appropriate size.
