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Microbial transformation

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Abhishek Rana

This document discusses biotransformation and microbial conversion of organic compounds. It defines biotransformation as the microbial conversion of a substrate into a product using enzymatic reactions. Various examples of biotransformation reactions are provided, including the production of gluconic acid, antibiotics, lactic acid, and acetic acid (vinegar). The document also discusses methods for biotransformation like growing, resting, and immobilized cells as well as cell-free extracts and enzymes. It provides details on the production processes of certain compounds like gluconic acid, vinegar, and steroids.

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STUDENT RKGIT (PHARMACY) Dr. A.P.J. Abdul Kalam Technical University
Microbial transformation
Transformation Biotransformation Types of reactions mediated by microorganisms Biotransformation reactions Selection of organisms Methodologies for Biotransformation Process improvements with special reference to steroids
Definition Transformation is defined as the genetic alteration of cell resulting from the direct uptake and incorporation of exogenous genetic material from its surroundings through the cell membrane
Definitions :- It refers to the process in which microorganisms convert organic compounds into structurally related products. or It deals with microbial(enzymatic) conversion of a substrate into a product with limited number enzymatic reactions.
Gluconic acid production Antibiotics production Lactic acid production Single cell protein from methanol Acetic acid (vinegar) production Kojic acid Itaconic acid
It possess applications in a variety of product and product utilities in pharmaceuticals such as :- Calcium Gluconate to provide and supplement Ca to pregnant mothers Na and Ca salts in alkaline serve as effective metal sequestering agents for Fe , Al Ferrous gluconate caters for Fe for the treatment of Anaemia in humans Sodium gluconate finds its use as a sequestering agent in detergents
Fermentation Production The production of gluconic acid from glucose is carried ot by flavoprotein known as glucose oxidase.
The production of vinegar on commercial scale may be accomplished by several tests as stated below:- Aerobic fermentation process Submerged fermentor Packed generator process Orleans process Traditional method
In production of vinegar it requires two different modes of fermentations ,namely a) Utilyzing a apecific yeast to produce ethanol from cane sugar b) Utilyzing Acetobacter species to carry out oxidation of ethanol The vinegar production is initiated from the alcoholic broth by following steps:- alcohol content adjusted between 10-13% Vinegar in small quantity added to enhance the acidity of broth Alternatively, Acetobacter cells may incorporated in such process that would require inoculation.
Microbial transformation
Microbial transformation
The selection of strains from its natural sources or from available cultures are responsible for catalyzing the desired biotransformation reactions. It can be done by following methods:- 1. Modified Enrichment Method 2. Filtration Enrichment Method 3. Agar plug method
It is used for the isolation of mutants blocked in the substrate dissimilation mechanism. In this a steroid substrate is incorporated as the sole C-source exclusively in a minimal medium seeded adequately with the soil dilutions.The cells that causes the degradation of the substrate will grow; and therefore,subsequently transferres to the same medium but enriched with another C source : glucose. However , the mutants present which are blocked at different stages in the process of degradation of steroid substrate , consume glucose as the C source.
In this case , after mutagenesis the spores of filamentous organisms e.g.,actinomycetes, fungi, are made to develop in a liquid minimal medium. The micro-colonies of prototrophs thus developed are meticulously separated by filtration, where by the spores of auxotrophs that were unable to grow left behind in the filtrate. The filtrate obtained in this manner in subsequently plated and the resulting colonies are adequately checked for
In this agar plates are inoculated with a standardized inoculum of the test microorganism. Then, filter paper (about 6 mm in diameter), containing the test compound at a desired concentration, are placed on the agar surface. The Petri dishes are incubated under suitable conditions. Generally, antimicrobial agent diffuses into the agar and inhibits germination and growth of the test microorganism and then the diameters of inhibition growth zones are measured.
The methodolgis for biotransformation includes:- Growing cells Resting cells Killed cells Immobilized cells Cell-free extract, Enzymes Immobilized enzymes
The desired cells are cultivated in a suitable medium. As the growth of the cell soccurs (6-24hours), a concentrated substrate is added to the culture. Sometimes ,addition of emulsifiers (Tween, organic solvents) is required to solubilize substrates and/or products eg . Steroid biotrasformation. The substrate conversion to product can be monitored by spectroscopic or chromatographic techniques. Biotransformation can be terminated when the product formation is optimum.
In such critical situations when the enzyme induction afforded by the added substrate is not quite necessary and urgent, resting cells may be employed profusely and effectively. However, the resting cells do offer a tremendous advantage where by the growth inhibition by the substrate is eliminated completely. Besides, the presence of high-cell densities that essentially promote an enhanced level of productivity may be employed ; simultaneously, the very risk of any possibles cope of contamination is minimised appreciably. Interestingly, there are several biotransformation reactions that exclusively and predominantly take
Biotransformation can be carried out continuously by employing immobilized cells. Further, the same cells could be used for numerous time. Several bioconversion with single or multistage reaction are in fact carried out by using immobilized cells . Eg. Commercial production of L-analine and malic acid.
Naturally occurring steroids possess remarkable hormonal properties which are of therapeutic importance to human well- being, such as hormones of adrenal cortex (cortisone, cortisol ,corticosterone), the progestational hormone (progesterone), the androgens or male sex hormones(testosterone, di hydrotestosterone) and the estrogens or female sex hormones (estradiol, estrone, etc.)
Microbial transformation
Microbial transformation

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Microbial transformation

  • 1. STUDENT RKGIT (PHARMACY) Dr. A.P.J. Abdul Kalam Technical University
  • 3. Transformation Biotransformation Types of reactions mediated by microorganisms Biotransformation reactions Selection of organisms Methodologies for Biotransformation Process improvements with special reference to steroids
  • 4. Definition Transformation is defined as the genetic alteration of cell resulting from the direct uptake and incorporation of exogenous genetic material from its surroundings through the cell membrane
  • 5. Definitions :- It refers to the process in which microorganisms convert organic compounds into structurally related products. or It deals with microbial(enzymatic) conversion of a substrate into a product with limited number enzymatic reactions.
  • 6. Gluconic acid production Antibiotics production Lactic acid production Single cell protein from methanol Acetic acid (vinegar) production Kojic acid Itaconic acid
  • 7. It possess applications in a variety of product and product utilities in pharmaceuticals such as :- Calcium Gluconate to provide and supplement Ca to pregnant mothers Na and Ca salts in alkaline serve as effective metal sequestering agents for Fe , Al Ferrous gluconate caters for Fe for the treatment of Anaemia in humans Sodium gluconate finds its use as a sequestering agent in detergents
  • 8. Fermentation Production The production of gluconic acid from glucose is carried ot by flavoprotein known as glucose oxidase.
  • 9. The production of vinegar on commercial scale may be accomplished by several tests as stated below:- Aerobic fermentation process Submerged fermentor Packed generator process Orleans process Traditional method
  • 10. In production of vinegar it requires two different modes of fermentations ,namely a) Utilyzing a apecific yeast to produce ethanol from cane sugar b) Utilyzing Acetobacter species to carry out oxidation of ethanol The vinegar production is initiated from the alcoholic broth by following steps:- alcohol content adjusted between 10-13% Vinegar in small quantity added to enhance the acidity of broth Alternatively, Acetobacter cells may incorporated in such process that would require inoculation.
  • 13. The selection of strains from its natural sources or from available cultures are responsible for catalyzing the desired biotransformation reactions. It can be done by following methods:- 1. Modified Enrichment Method 2. Filtration Enrichment Method 3. Agar plug method
  • 14. It is used for the isolation of mutants blocked in the substrate dissimilation mechanism. In this a steroid substrate is incorporated as the sole C-source exclusively in a minimal medium seeded adequately with the soil dilutions.The cells that causes the degradation of the substrate will grow; and therefore,subsequently transferres to the same medium but enriched with another C source : glucose. However , the mutants present which are blocked at different stages in the process of degradation of steroid substrate , consume glucose as the C source.
  • 15. In this case , after mutagenesis the spores of filamentous organisms e.g.,actinomycetes, fungi, are made to develop in a liquid minimal medium. The micro-colonies of prototrophs thus developed are meticulously separated by filtration, where by the spores of auxotrophs that were unable to grow left behind in the filtrate. The filtrate obtained in this manner in subsequently plated and the resulting colonies are adequately checked for
  • 16. In this agar plates are inoculated with a standardized inoculum of the test microorganism. Then, filter paper (about 6 mm in diameter), containing the test compound at a desired concentration, are placed on the agar surface. The Petri dishes are incubated under suitable conditions. Generally, antimicrobial agent diffuses into the agar and inhibits germination and growth of the test microorganism and then the diameters of inhibition growth zones are measured.
  • 17. The methodolgis for biotransformation includes:- Growing cells Resting cells Killed cells Immobilized cells Cell-free extract, Enzymes Immobilized enzymes
  • 18. The desired cells are cultivated in a suitable medium. As the growth of the cell soccurs (6-24hours), a concentrated substrate is added to the culture. Sometimes ,addition of emulsifiers (Tween, organic solvents) is required to solubilize substrates and/or products eg . Steroid biotrasformation. The substrate conversion to product can be monitored by spectroscopic or chromatographic techniques. Biotransformation can be terminated when the product formation is optimum.
  • 19. In such critical situations when the enzyme induction afforded by the added substrate is not quite necessary and urgent, resting cells may be employed profusely and effectively. However, the resting cells do offer a tremendous advantage where by the growth inhibition by the substrate is eliminated completely. Besides, the presence of high-cell densities that essentially promote an enhanced level of productivity may be employed ; simultaneously, the very risk of any possibles cope of contamination is minimised appreciably. Interestingly, there are several biotransformation reactions that exclusively and predominantly take
  • 20. Biotransformation can be carried out continuously by employing immobilized cells. Further, the same cells could be used for numerous time. Several bioconversion with single or multistage reaction are in fact carried out by using immobilized cells . Eg. Commercial production of L-analine and malic acid.
  • 21. Naturally occurring steroids possess remarkable hormonal properties which are of therapeutic importance to human well- being, such as hormones of adrenal cortex (cortisone, cortisol ,corticosterone), the progestational hormone (progesterone), the androgens or male sex hormones(testosterone, di hydrotestosterone) and the estrogens or female sex hormones (estradiol, estrone, etc.)