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PolyHydroxyButyrate From Whey

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by

Muhammad Usama

on 5 March 2015

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Transcript of PolyHydroxyButyrate From Whey

Dust particles=278.05-272.49=5.56 ton
Group Members :
Muhammad Usama 2011-uet-iefr/chem/fd-77
Production Of PHB from Whey
Muhammad Zia - Ul - Haq
Zaid Yahya 2011-uet-iefr/chem/fd- 100
Awais Ali Sultani 2011-uet-iefr/chem/fd- 53
Nasir Ali 2011-uet-iefr/chem/fd- 49
Shahryar Rafaqat 2011-uet-iefr/chem/fd-73
Faizan Qurban 2011-uet-iefr/chem/fd- 47
Group Advisor :
(Assistant Professor)
Biodegradable Plastic)
( PHB : PolyHydroxyButyrate )
Ultra-filtration unit
47256.22=4725.62+42530.60
47256.22=47256.22
Input=Output
Evaporator
42530.60 = 35695.32 + 6835.28
42530.60 = 42530.60
Input = Output
Culturing Tank
1270.34 + 522.91 = 1793.25
1793.25 = 1793.25

Input = output
Blending Tank
1793.25+6835.28=8628.53
8628.53=8628.53
Input=output
Seed Fermenter
The seed fermenter is used to inoculate the production fermenter.
Sodium hydroxide is added to control the pH of the culture broth.
1913.88+316.92+170.88=1913.88+8762.55+134.69+562.53+219.411+170.88
11763.94 ton=11763.94 ton
Input=Output
Seed Fermenter
Fermenter
The general equation of PHB production is
C; 12 = c + 4d + e →(1)
H; 22 + 3b = 1.77c + 6d + 2f →(2)
O; 11 + 2a = 0.49c + 2d + 2e + f →(3)
N; b = 0.24c →(4)
Applying Elemental balance;
11763.94+84.89=278.05+11570.78
11848.83 ton=11848.83 ton

Input=output
Streams Codes and Description

S-22 Filtered air from A-17 is discharged to Atmosphere
S-23 Product after Fermentation is sent to Storage Tank A-18
S-24 unrefined PHB is sent to Centrifuge A-19
S-25 Aqueous waste is separated from Centrifuge A-19
S-26 Product of Centrifuge A-19 is sent to Blending Tank A-21
S-27 Surfactant (A-20) is added to Blending Tank A-21
S-28 Stream is sent to the Pump A-22
S-29 Stream from Pump A-22 to the Mixer A-25
S-30 Hypochlorite (A-23) is sent to Pump A-24
S-31 Pumped Hypochlorite is sent to Mixer A-25
S-32 Product Of Mixer (A-25) is sent to Centrifuge A-26
S-33 Aqueous waste from Centrifuge A-26 is discharged
S-34 Product of Centrifuge A-26 is sent to Blending Tank A-28
S-35 Water (A-27) is added to Blending tank A-28 for washing
S-36 Stream from Blending Tank A-28 is sent to Centrifuge A-29
S-37 Aqueous Waste from Centrifuge A-29 is discharged
S-38 Product of Centrifuge A-29 is sent to Spray Dryer A-30
S-39 Air is supplied to Spray Dryer A-30
S-40 Air leaves the Spray Dryer A-30
S-41 Product of Spray Dryer is sent to Storage Tank A-31
FlowSheet
S Whey from the Dairy to Storage Tank A-01
S-1 Whey sent to Ultra-Filteration Unit A-02
S-2 Whey retenate is rejected
S-3 Whey permeate is sent to Evaporator A-03
S-4 Steam is supplied to Evaporator A-03
S-5 Vapors from Evaporator A-03
S-6 Steam Condensate from Evaporator A-03
S-7 Thick liquor is sent to Blending tank A-07
S-8 Supplements from A-04 to Culture tank A-06
S-9 Ammonium Sulphate from A-05 to Culture Tank A-06
S-10 Culturing Medium is sent to Blending Tank A-07
S-11 Culture (A-07) is sent to Heat Sterilizer A-08
S-12 Culture (A-08) is sent to Cooler A-09
S-13 Culture (A-09) is fed to the Fermenter Seeder A-12
S-14 NaOH Solution (A-10) is fed to Fermenter Seeder A-12
S-15 E.Coli Cells (A-11) fed to the Fermenter Seeder A-12
S-16 Stream is sent to Fermenter A-16
S-17 Air is supplied to Compressor A-13
S-18 Compressed air is sent to Air Filter A-14
S-19 Compressed Filtered air is sent to Cooler A-15
S-20 Compressed Filtered cooled air is fed to fermenter A-16
S-21 Air sent to Filter A-17
Basis : 1 Day Operation
Whey Requirement
The components in 1 liter whey solution are:
So, in 47256.22 ton whey solution:

Lactose amount=2126.53 ton

Water amount=47256.22 ton*0.937
Water amount=44279.08 ton

Protein amount=47256.22 ton*0.008
Protein amount=378.05 ton

Salt amount=0.01*47256.22 ton
Salt amount=472.56 ton

Desired capacity=600 ton of PHB


PHB amount=600 ton of PHB*1/0.95
PHB amount=631.58 ton of PHB
Yield=0.33 (tons of PHB)/(tons of lactose)
Recovery of PHB after fermentation=95%
Total Required Lactose
After Ultrafiltration 90% of lactose is recovered from whey solution
Total Required Lactose=2126.53 tons
Whey requirement
Whey requirement=47256222.22 liter
Whey requirement = 47256.22 Ton/day
Sterilizer
Cooler
Molecular weight of PHB = 86 g/gmol
Molecular weight of biomass = 24.97 g/gmol
Molecular weight of lactose = 342 g/gmol
Mass in + Mass generated = Mass out + Mass consumed
For total mass and Nitrogen the mass balance equation is

Mass in = Mass out

(at the inlet of fermenter)
Air Filter
Dust particles=86.62-84.89=1.73 ton
(at the outlet of fermenter)
Air Filter
Storage Tank
Centrifuge (A-19)
11570.78=9246.78+2324
11570.78 ton=11570.78 ton
Input = Output
Blending Tank (A-21)
2324 ton+232.4 ton=2556.4 ton
2556.4 ton=2556.4 ton
Input=output
Pump (A-22)
Pump (A-24)
Mixing Tank (A-25)
2556.4 + 33.58 = 2589.98
2589.98 ton = 2589.98 ton
Input = Output
2589.98=864.55+1725.43
2589.98 ton=2589.98 ton
Input=Output

Centrifuge (A-26)
Blending Tank (A-28)
864.55+12.79=877.34
877.34 ton=877.34 ton
Input=Output
Centrifuge (A-29)
877.34=857.14+20.20
877.34 ton=877.34 ton
Input=Output
Spray
Dryer
47256.22+
522.91+
1270.34+
170.88+
2964.55+
86.62+
232.4+
33.58+
12.79+
16382.20
4725.62+
35695.32+
278.05+
1.73+
9246.8+
1725.43+
20.20+
16627.10+
612.24
68932.49 ton = 68932.49 ton
Input = Output
Inputs
Outputs
Evaporator
Mixing tank
Heat
Sterilizer
Cooler
Seed Fermentor
Compressor
Cooler
Fermenter
Boiler
PUMP (A-22)
Pump (A24)
Spray
Dryer
Reference:
Design and techno-economic evaluation of microbial biopolymer production from food industry waste and agricultural crops
Isabel Lopez Garcíaa, M. Pilar Dorado Pereza, Jimmy A. Lópezb, Marcelo A. Villarb, Stavros Yanniotisc and Apostolis Koutinas
Reference:
Optimization of Bioplastics Production from Cheese Whey
(Inês da Silva Farinha)
Bioprocess Engineering Principles by
Pauline M. Doran, 3rd edition, chapter # 4
Production of Poly(3-Hydroxybutyrate) by Fed-Batch Culture of Recombinant E. Coli with a Highly Concentrated Whey Solution
Woo Suk Ahn, Si Jae Park, and Sang Yup Lee*
Reference:
Reference:
Spray drying by
J.weernink
Reference:
University botany 1 by
S.M Reddy Vol 1 Page 205
Process analysis and economic evaluation for Poly hydroxy butyrate production by fermentation

Jong-il Choi, Sang Yup Lee
Design and techno-economic evaluation of microbial biopolymer production from food industry waste and agricultural crops
Isabel Lopez Garcíaa, M. Pilar Dorado Pereza, Jimmy A. Lópezb,
Marcelo A. Villarb, Stavros Yanniotisc and Apostolis Koutinas
Bioprocess Engineering Principles by
Pauline M. Doran, 3rd edition, chapter # 4
Spray drying by
J.weernink

Production of Poly(3-Hydroxybutyrate) by Fed-Batch Culture of Recombinant Escherichia coli with a Highly Concentrated Whey Solution

Woo Suk Ahn, Si Jae Park, and Sang Yup Lee*
Optimization of Bioplastics Production from Cheese Whey
Inês da Silva Farinha
University botany 1 by
S.M Reddy Vol 1 Page 205
Chemical Process Principles By
O. A. and Watson, K. M. Hougen

Basic Principles and Calculations in Chemical Engineering
By David M. Himmelblau
References
Purification and Recovery Section
Production of polyhydroxybutyrate from wheat based feed stock...
The university of Manchester
Process analysis and economic evaluation for Poly hydroxy butyrate production by fermentation
Jong-il Choi, Sang Yup Lee
Reference:
Energy Balance
References:
1. http://pubchem.ncbi.nlm.nih.gov/compound/24436#section=Top (Open Chemistry Database)
2. http://www.ncbi.nlm.nih.gov/ ( Open Database)
3. http://highered.mheducation.com/sites/dl/free/0072849606/315014/physical_properties_table.pdf (Compiled from data in Perry’s Chemical Engineers’ Handbook, 6th ed., Biochemical and Biotechnology Handbook, 1991, 2nd ed. and Process Synthesis, D. F. Rudd, G. J. Powers and J. J. Siiroia, 1973.)
4. http://courses.chem.indiana.edu/c360/documents/thermodynamicdata.pdf
5. Materials and the Environment: Eco-informed Material Choice, 2nd Edition, Chap. 15 Material Profiles Pg#508 (By M. F. Ashby)
6. http://webbook.nist.gov/cgi/cbook.cgi?ID=C7757826&Type=JANAFS&Plot=on#JANAFS (Chemistry web book )
7. http://fblgroup.com.pk/steam-gen-cost10fuels/
Enthalpy and Heat Data at Specific Temperature
Heat Capacity & Enthalpy Data
at Specific Temp.
Heat Capacities of Components at Specific Temperature
Production of PHB by the fermentation of dairy
waste(Whey) with the help of E.Coli

Introduction

[1]http://oceancrusaders.org/crusades/plastic-crusades/plastic-statistics

It is now believed that there are 5.25 trillion pieces of plastic debris in the ocean. Of that mass, 269,000 tons float on the surface, while some four billion plastic microfibers per square kilometer litter the deep sea.[1]
APPROXIMATELY 1 MILLION SEA BIRDS ALSO DIE FROM PLASTIC.[1]
Plastic buried deep in land fills can leach harmful chemicals that can spread into ground water.
The solution is to use something degradable and bearable to our environment.
And the answer is Biodegradable plastic.
Need of Biodegradable Plastic
Importance:
The amount of plastic waste increases every year and the exact time needed for its biodegradation is unknown. Nowadays plastics and synthetic polymers are mainly produced using petrochemical materials that cannot be decomposed.(Hazardous)
it is 100% degradable.
Requires no petrochemicals therefore no bad environmental impacts.
PHB
What is PHB?[1]
Polyhydroxybutyrate (PHB) is a polyhydroxyalkanoate (PHA), a polymer belonging to the polyesters class that are of interest as bio-derived and “biodegradable plastics”. It is a thermo plastic material.
[1]Biosynthesis, biodegradation, and application of poly(3-hydroxybutyrate) and its copolymers - natural polyesters produced by diazotrophic bacteria
[2] Ahn W.S., Park S.J. & Lee S.Y. 2000. Production of Poly(3-hydroxybutyrate) by Fed-Batch Culture of Recombinant Escherichia coli with a Highly Concentrate Whey Solution. Applied and Environmental Microbiology, 66(8), 3624–3627.
[3] A Handbook of Applied Biopolymer Technology: Synthesis, Degradation and Applications by Sanjay Kumar Sharma, Ackmez Mudhoo, James H. Clark. Page 316
Synthesis:
Nitrogen-fixing microorganisms can synthesize PHB in considerable quantity.[1]
Recombinant strain of E. coli CGSC 4401, harboring the plasmid pJC4 containing the Alcaligenes latus PHA biosynthesis genesis used in our project.[2]
Biodegradation:
PHB serve as storage compound for carbon under growth limiting conditions. Thus can be utilized by microorganisms.[3]
Degrades to H2O and CO2.
Uses:
Medical Industry
In the pharmacology it can be used as drug carrier.
In food industry as bottles.
PHB
[1]Ahn W.S., Park S.J. & Lee S.Y. 2000. Production of Poly(3-hydroxybutyrate) by Fed-Batch Culture of Recombinant Escherichia coli with a Highly Concentrate Whey Solution. Applied and Environmental Microbiology, 66(8), 3624–3627.
Choi J., & Lee S.Y. 1999. Factors Affecting the Economics of Polyhydroxyalkanoates Production by Bacterial Fermentation. Applied Microbiology and Biotechnology, 51(1), 13-21.

It provides us the Carbon source i.e Lactose.
It is a by-product of the manufacture of cheese.
While making 1 kg of cheese 9 litters of whey is generated.
Composition of whey:[1]
Whey
Process description:
In this process PHB is produced by Fermentation of Lactose(Carbon Source) with the help of E.Coli.
Ultrafiltration is done for the protein and salt removal.
Evaporator is used to concentrate the whey solution.
Then it is mixed with Ammonium Sulphate(nitrogen source) & mineral supplements.
Sterilization is required to remove undesired Microorganisms.
Then Fermentation occurs with the help of E.Coli.
In the downstream process the PHB is purified.
PHB remaining in cells is extracted and purified via Centrifugation, surfactants-hypochlorite digestion and water washing processes respectively.
[1]Choi J. & Lee S.Y. 1999. Efficient and Economical Recovery of Poly(3-Hydroxybutyrate) from Recombinant Escherichia coli by Simple Digestion with Chemicals. Biotechnology and Bioengineering, 62(5), 546-553.
[1]Choi J, & Lee S.Y. 1997. Process Analysis and Economic Evaluation for Poly(3-hydroxybutyrate) Production by
Fermentation. Bioprocess Engineering, 17(6), 335-342.
Capacity : 600 tons/Day
Material Balance
Overall Material Balance
Feed Purification and Preparation Section
Fermentation Section
Contents
Introduction
Flowsheet
Process Description
Raw Material Requirement
Material Balance
Energy Balance

Reason to use it:
The primary contributor to the overall cost is Carbon substrate cost which is up to 50%.and in our project its cost is negligible because it is taken from whey.
Easily available
Renewable
Economic Feasibility


Exports
Importance
Production Capacity

Conclusion
HEATS OF COMBUSTION OF HIGH TEMPERATURE POLYMERS
Richard N. Walters*, Stacey M. Hackett* and Richard E. Lyon
Reference:
Full transcript