Riboflavin Production- Biological Process
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A Biological process for the production of
Riboflavin
Team Riboflavulous
Team 1 members
Priyesh WaghmareYixue Chen
Rebecca MilburnMadhunika Padmanabha
Sharath Sathyan
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Outline:
IntroductionMarketChemical Vs biological methodEnvironmental impactMain processWaste treatmentMass balanceMerits of our systemFuture prospects
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Introduction to riboflavin (1)
Riboflavin ( vitamin B2)Molecular formula
C17H20N4OYellow-orange Sparingly soluble in
waterForms crystals in <30ºC
waterLight-sensitive
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Introduction to riboflavin (2)
Precursor of Co-enzymes - flavin adenine dinucleotide (FAD)
- flavin mononucleotide
Deficiency results in metabolic and skin disorders
Riboflavin is mainly used as a food supplement for both human and animals.
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Market
World demand is estimated to be at 6,000 tons p.a.
Major producers are Roche, BASF and China’s Hubei Guangji Pharmaceutical
Feed-grade (80% ) sells at US $30/kg pharmaceutical-grade (98%) sells at US
$50/kg.
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Chemical v/s Biological Process
Glucose
K arabonate
Ca arabonate
Ca ribonate
Ribonolactone
Ribose
Ribitylxylidine
Phenylazo
Riboflavin
Feed + Water
+Innoculum
Riboflavin
Chemical Process Biological Process
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Riboflavin production (Biological process)
Our design involves a single-step biological processRecombinant Bacillus subtilis is a gram positive,
aerobic bactiera that converts glucose directly into riboflavin
Recombinant B.subtilus can yield up to 16g/L riboflavin in 48hours
Riboflavin is sparingly soluble and forms crystals in the fermentation broth
Bacillus subtilis is much small than the produced riboflavin particles, making downstream purification easier
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Life cycle assessment (LCA) of Riboflavin Production
Type of process Chemical Biological
Raw materials (%) 100 150
Non-renewable raw materials(%)
100 25
Energy(%) 100 90
Emissions of VOCs(%) 100 50
Emissions to water(%) 100 33
(Organisation for Economic Co-operation and Development., 2001)
The biological method uses mainly renewable resources Less amounts of energy used in biological method Air and water are contaminated to a lesser degree by the biological process
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Environmental Fingerprint
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Overall Riboflavin Production Process
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Feed-grade production
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Pharma-grade production
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Material Recycling
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Energy Recycling
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Waste Treatment – HCL gas
• HCL gas emitted in the process is removed by scrubbers.
• In the packed tower, HCL gas flows upward through a packed bed (provides close gas-liquid contact) while the scrubbing liquid flows downward by gravity over the packing.
• The internal components of the tower consist of a packing support plate, a packed bed, a liquid distributor and a mist eliminator.
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Waste Treatment – Biomass waste
Biodegradable organic waste is collected and put into an anaerobic digestion tank The material is broken down by bacteria in the absence of oxygen Biogas product is cleaned, compressed and sent to a CHP plant Solid digestate is also produced and can be
used as fertilizer
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Used Charcoal- Thermal regeneration process
Adsorbent drying at approximately 105 °CDesorption and decomposition at 500–
900°C under an inert atmosphereResidual organic gasification by carbon
dioxide at 800°C
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Mass Balance
Fermentation
Feed (3.3)
Inoculums (5)
Water 2.5
Air 30
Nitrogen 22.6
Oxygen 5.2CO2 3.65
Waste Water 7.35
98.55
Differential Centrifugation
Spent Biomass 4.7
Water 84.5
Impure Riboflavin Crystals 2All Measurements in tones
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Mass Balance
Impure Riboflavin Crystals 2 Acid Wash
Impurities (0.4)
Dilute HCL 1.4
Dilute HCL 0.2
Drum Dryer
Riboflavin 1.8
Acid + Water+ Trace amounts Biomass 0.2
Feed Grade
Riboflavin 1.61.6
All Measurements in tones
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Differential Centrifugation:
Foodec decanter centrifuges focus on Hygiene Reliability Easy access User friendly Low noise level
Alfa Laval Foodec decanter centrifuges - used for pharmaceutical applications that comply with strict sanitary regulations.
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Sizing & Cost:
Fodec 800
Capacity Depends on application
G-force max 3243
Bowl material Duplex stainless steel
Other weight parts
AISI 316
Weight Kg 13000 (28860 lbs)
Installed Power kW 132-250 (140-330Hp)
Sound Pressure Level
dB(A) re.20lpa 89
Cost 30,000 US Dollars
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Downstream Processing Equipments:
Candle filter System:•Particulate removal system.•Ceramic Filters•Cyclones•Residues up to 1ppm removed
Stirred Tank Fermenter:•Coiled tube for Heating & cooling•Uniform mixing•Better fermentation•Defoamer
Candle Filter System:
Stirred Tank Fermenter:
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Merits of our system
A simple one-step biological processBacillus subtilis requires a relatively
unrefined growth conditionsNo harmful chemicals used in process>90% culture medium recycle>90% hydrochloric acid recycleUsed biomass recycling as nutrientsEnergy recycling system in placeLarge storage tank in place to make full
usage of downstream process equipment
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Future Prospects
To genetically modify bacillus subtilis to create a strain that has a higher product yield (>16g/L)
To replace part of the glucose feed with cheaper organic residues like rotten potatoes/oranges
To develop a process for purification of pharma-grade riboflavin with fewer steps