Microbial Biotransformation for B. Pharm student as per PCI syllabus

1. Microbial Biotransformation
By:
Vasundhara S. Kakade
Assistant Professor
NES Navsahyadri Institute of Pharmacy, Nasarapur Pune.
3/30/2020 1NES NIPT.Y. B.Pharm

2. Introduction
• Biotransformation ( Bio+ transformation)
• Living process which carry changes in organic compound.
• biological processes in which microorganisms convert organic
compounds into structurally modified reusable products.
• In other word, biotransformation deals with microbial
(enzymatic) conversion of a substrate into a product with limited
number by enzymatic reactions.
• This is in contrast to fermentation which involve a large number
reactions e.g. steroids produced by chemical synthesis or by
microbial biotransformation. This are physically active
compounds which includes testosterone, progesterone etc. act as
bio regulator in body.
3/30/2020 2NES NIPT.Y. B.Pharm

3. Advantages
This process is carried out by enzyme secreted by m.o.
• React specifically
• No side reactions
• Specific micro-organism used to carry out specific type of
transformation.
• It can be carried out mild condition like neutral pH , RT &
at normal pressure.
• Reduce multistep chemical reaction.
• Occurs in less labour, time, money, and release good yeild
• Less envt. pollution
3/30/2020 3NES NIPT.Y. B.Pharm

4. Methods in Biotransformation
• Microbial cell play major role in biotransformation. Major
step is fermentation.
• It is carried out in following phases
Phase I: Growth Phase
Phase II :Transformation Phase
Phase I: Growth Phase
 Culture is grown in medium, provided with areation, agitation,
with optimum temperature for growth.
 Incubated for few days depend on type of micro-organism.
3/30/2020 4NES NIPT.Y. B.Pharm

5. • Phase II :Transformation Phase
 Transformation phase starts by addition of steroid at end of
growth phase. With incubation period, amount of steroid depends
on capacity of culture, toxicity & type of product .
 Enzymes secreted by micro-organism acts on steroids. And
transformation occurs depend on temp, pH, agitation, aeration
and incubation time.
3/30/2020 5NES NIPT.Y. B.Pharm

6. General Process for biotransformation of steroids carried out in
stainless steel tank
Prepare media to
carry out high
extraction &
purification of
transform product
Add steroids to start
process of
transformation
require 24-48 hrs for
incubation
After transformation
micro-organism
separated from culture
media & extracted with
solvent. (CHCl3, ethyl
acetate etc .) by solvent
extraction &
precipitation method
Extract samples separated properly
& analyzed by TLC, Paper
chromatography ,GC or HPLC
And then new structure is identified
by IR, NMR, Mass spectroscopy
3/30/2020 6NES NIPT.Y. B.Pharm

7. Types of Biotransformation reactions
• Many types of chemical reactions occur in
biotransformations.
•These include various reactions like oxidation, reduction,
hydrolysis, condensation, isomerization, formation of new C-C
bonds, synthesis of chiral compounds and reversal of
hydrolytic reactions.
•Among these, oxidation, isomerisation and hydrolysis
reactions are more commonly occur.
3/30/2020 7NES NIPT.Y. B.Pharm

8. Various applications of microbial
biotransformation
• Steroids constitute a natural product class of compounds
that is widely distributed throughout nature present in bile
salts, adrenal-cortical and sex-hormones, insect molting
hormones, sapogenins, alkaloids and in some antibiotics.
• Testosterone was produced from dehydroepiandrosterone
by using Corynebacterium sp.
• Subsequently cholesterol was produced from 4-
dehydroeticholanic and 7-hydroxycholestrol using Nocardia
spp..
3/30/2020 8NES NIPT.Y. B.Pharm

9. All steroids have the same basic structure, a
cyclopentanoperhydrophenanthrene which consists of four fused rings
• Cortisone and prednisolone is very useful because of its anti-
inflammatory action against rheumatoid arthritis and skin diseases.
By changing the structure, specifically by incorporating a 1,2 double
bond in ring A of the cortisone molecule to produce prednisone have
anti-inflammatory with the help of an enzyme named as Arthrobacter
simplex.
3/30/2020 9NES NIPT.Y. B.Pharm

10. • Transformation of non-steroid compounds
Dihydroxyacetone from glycerol
• It is used in lotions and cosmetics. Various acetic acid
bacteria such as Gluconobacter melanogenus used in this
microbial transformation.
Prostaglandins
• unsaturated fatty C-20 fatty acids hormones.
• used as contraceptive, alleviation of pain of child-birth,
treatment of congenital heart failure and also for the
treatment of digestive diseases.
• These prostaglandins can be produced from unsaturated
fatty acids by microbial transformation with pathogenic fungi
such as Cryptococcus neoformans.
3/30/2020 10NES NIPT.Y. B.Pharm

11. • Transformation of antibiotics
Develop modified and improved antibiotics with reduced toxicity,
broad antimicrobial spectrum, enhanced oral adsorption, allergic
effects
• Transformation of pollutants
biotransformation of various pollutants carried out by using
microorganisms to clean up the polluted environment.
• Petroleum biotransformation
Petroleum is important for industry and daily life. Now a day
hydrocarbon contamination resulting from the activities related to the
petrochemical industry. Leaks and accidental release are the main
source of soil and water pollution. Petroleum oil is carcinogenic and
neurotoxic for all living forms.So various mechanical and chemical
methods have been employed for soil and water remediation. But,
bioremediation using microbes is the most efficient method to
detoxify the pollutants because it leads to complete mineralization
and is cost effective.
3/30/2020 11NES NIP T.Y. B. Pharmacy

12. References
• BY C.R. Kokare Pharmaceutical Biotechnology
Nirali Prakashan.
• By U satayanarayana & Chakrapani
Pharmaceutical Biotechnology International
Edition.
3/30/2020 NES NIPT.Y. B.Pharm 12

13. QUESTIONS
1. Define microbial biotransformation with its process . 5 M
2. Give application of microbial biotransformation 5M
• THANK YOU
3/30/2020 NES NIPT.Y. B.Pharm 13