PHB PRODUCTION USING BANANA
PEEL FOR MEDICAL USE
Under the supervision of Dr Ashok Srivastava
01
INTRODUCTION
Biopolymers are polymers produced from
natural sources either chemically synthesized
from a biological material or entirely
biosynthesized by living organisms.
TYPES- PLA, PHA, PHB, PHV
PHB is considered best replacement for
petroleum based plastics
02
03
My objective for this project will
be high yield production of
medical grade PHB using
banana peel. OBJECTIVE
Secondly, finding out if
traditional medical materials
can be replaced by
biopolymers or not .
Literature Review
There are many researches in the field of PHB
production using gram negative bacteria, but
still the PHB produced is too expensive to
compete with petroleum based plastic.
For my project, I will be using gram positive
bacteria (Bacillus thuringiensis) for production of
non toxic PHB, also use of banana peel which
helps in better waste management and makes the
process cost effective.
04
08
Why Bacillus
thuringiensis?
Gram positive bacteria
Accumulates PHB up to 60.07% of
cellular dry weight
Natural PHB producer
Spore Formation
Robust growth
Safety
Why banana peel?
Banana peel is one of the abundant waste materials
in India.
It is high in organic matter (91.5%), including
carbohydrate (59%), protein (0.9%), and crude fat
(1.7%), as well as minerals like potassium (0.78%),
calcium (0.19%), sodium (0.24%), iron (0.006%), and
manganese (0.76%), all of which stimulate bacterial
growth.
To reduce the cost, banana peels will be
utilized as a raw material.
06
Preparation of medium and
activation of Bacillus
thuringiensis.
Inoculation of Bacillus
thuringiensis and
WORK PLAN incubation.
Extraction of PHB.
Determination of PHB
07
Methodology
1. bacillus thuringiensis is obtained and Stock culture will be maintained on
slants of nutrient medium at 4 °C
2. Growth-associated PHB production will be carried out in a banana peel
medium. Fresh banana peels can be collected from student centre and near
by juice shops.
3. These will be first washed thoroughly to clear the dirt and shredded into
small pieces. All the shredded pieces were dried in hot air oven at 50 °C for
24 h and 30 g of the peel was boiled in 250 mL distilled water at 100 °C for 40
min.
4. Obtained mixture was blended and the suspension will be treated with
H2SO4 (0.1 N) and tricalcium phosphate (2%w/v) for 4 h. Then, this mixture
will be centrifuged at 3000 rpm for 20 min. pH of the supernatant will be
adjusted to 7 by using sodium hydroxide (1 N).
5. The MSM (mineral salt medium) consisting of 2.0 g/l KH2PO4 and 0.2 g/l
MgSO4 will be used throughout the work to formulate the culture medium.
6. The final volume of the medium will be made 1000 mL by using distilled
water.
08
Methodology
7. The extraction of PHB will be done by following the protocol with some modifications-
A. After 72 h of growth in carbon rich medium at 35 °C, 100 mL of the broth will be
collected and centrifuged at 8000 rpm for 10–15 min.
B. Now the pellet will be treated with 10 mL sodium hypochlorite and kept at room temp
for half an hour.
C. The content will be again centrifuged at 10,000 rpm for 30 min.
D. The supernatant will be discarded and the pellet will be washed twice with ethanol and
acetone to rupture the cell wall and remove the unwanted cell debris.
E. The PHB powder will be extracted with boiling chloroform and filtered through
Whatman’s No 1 filter paper.
F. This will be followed by addition of 10 mL conc. sulphuric acid and kept at 100 °C for 10–
15 min in water bath.
8. PHB will be estimated spectrophotometrically by recording absorbance at 235 nm
against sulphuric acid blank.
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Problem to be addressed
Poor yield of PHB with Use of non
gram positive bacteria biodegradable medical
material in body
how can we improve the
yeild? how can we replace
traditional materials with
PHB?
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HARSH SINGH
2020UBT1058
THANK YOU
