enzyme and ethanol production using water hyacinth biomass
DESCRIPTION
Powerpoint presentation describing the possibility for using water hyacinth biomass as as feedstock for production of biofuels and biomass hydrolyzing enzymesTRANSCRIPT
Rajeev K Sukumaran, PhDScientist, Biotechnology Division
National Institute for Interdisciplinary Science and TechnologyTrivandrum
Council of Scientific and Industrial Research (Govt. of India)
Enzyme and Bio-ethanol production using Water Hyacinth Biomass
The Potential and Prospects
National Institute for Interdisciplinary Science and Technology
Water Hyacinth Facts Botanical Name: Eichornia crassipes
Common Name : Common Water Hyacinth (One of the seven species of the genus Eichornia)
Free floating perennial aquatic plants native to South America
Introduced in other parts of the world as ornamental plants and later became one of the worst aquatic weeds
Reproduces either through runners (stolons) which eventually form daughter plants or through seeds
Water Hyacinth is one of the most productive plants on earth
It can double in biomass in as little as 6-15 days
An acre of water hyacinth can weigh more than 200 tons
When not controlled, water hyacinth will cover lakes and ponds entirely blocking waterways and hampering all aquatic activity
This can impact water flow, block sunlight from reaching native aquatic plants, and starves the water of oxygen often leading to fish kills.
The plants also create a prime habitat for mosquitoes and a species of snail known to host a parasitic flatworm which causes schistosomiasis (snail fever)
National Institute for Interdisciplinary Science and Technology
WATER HYACINTH FACTS
Water ways infested with water hyacinth
The problems caused by excessive growth of the weed have attained devastating proportions in Kerala where inland water bodies are many
Massive invasion of paddy fields elicits a serious threat to paddy cultivation in Kuttanad area and in the districts of Ernakulam, Kottayam and Alappuzha
Many waterways in the state has become unserviceable due to the water hyacinth menace
Excessive infestation by the weed can severely constrain other activities like irrigation and fishing and prevent access to water navigation
The plant multiples rapidly in stagnant waters and sometimes can occupy the entire water surface from bank to bank stretching over 2-4 km along water channels
National Institute for Interdisciplinary Science and Technology
WATER HYACINTH MENACE IN KERALA
VietnamLake Victoria
Kenya
Florida
Philippines
WATER HYACINTH IS A GLOBAL PROBLEM !
National Institute for Interdisciplinary Science and Technology
CONTROL OF WATER HYACINTH
Chemical Control
Aquatic herbicides: 2,4-D, Diquat (6,7-dihydrodipyridol) and Glyphosate (low toxicity and rapid decomposition in water)
Methods are less expensive but serious ecological problem and human health hazard
Mechanical control
Choppers and shredders, mechanical harvesting, manual harvesting
Low efficiency, extremely expensive, energy intensive, perpetual practice needed. Probably suited only for small scale operations
Biological Control
Natural enemies of the plants employed (eg: Mite- Orthogalumna terebrantis Moth- Acigona infusella, Weevils- Neochetina eichhorniae, Neochetina bruchi, Fungi: Fusarium sp, Trichoderma sp
None of the methods are fully effective in controlling the weed !!
National Institute for Interdisciplinary Science and Technology
INTEGRATED CONTROL
No single method is effective in control of water hyacinth
An integrated control combines the different available methods chosen based on the region, type and level of infestation
National Institute for Interdisciplinary Science and Technology
INTEGRATED CONTROL WITH GENERATION OF ECONOMY
CONCEPT
Control measures should involve active participation from public and end benefactors
Active participation can come only through financial incentives
Remedial of water hyacinth should generate income and employment opportunities
Can the water hyacinth biomass removed from water bodies be in-fact beneficial ?
Isolated cases of benefits - utilization
Water hyacinth as animal feed/fodder
Generation of biogas
Production of compost/fertilizer
Water hyacinth as raw material (furniture, handicrafts, enzymes, alcohol)
National Institute for Interdisciplinary Science and Technology
ENZYME PRODUCTION FROM WH – A CASE FOR UTILIZATION
Water Hyacinth, as any other plant material contains cellulose, hemicellulose and lignin
Hemicellulose (40-50%)
Cellulose (12-18%)
Lignin (3-5%)
Cellulose is a polymer of glucose (a hexose sugar)
Hemicellulose is a polymer mainly containing arabinoxylan (Arabinose and xylose –pentose sugars)
Also contain Tannins
Microorganisms which can utilize these polymers for growth produces proteins which can degrade these polymers
The proteins which hydrolyze (breakdown) these polymers are enzymes
Cellulases , Xylanses, Tannases
National Institute for Interdisciplinary Science and Technology
ENZYME PRODUCTION FROM WH
Microorganisms –(Bacteria/Fungi) can grow on plant biomass and secrete these enzymes
Cellulases are a class of enzymes which find applications in
Textiles
Paper and pulp Industry
Food Industry
Laundry and Detergents
Biomass conversion and bio-ethanol production
World market estimated at 500 million US$
Xylanses : Applications in
Paper and pulp industry
Hemicellulose bioconversion and bioethanol production
National Institute for Interdisciplinary Science and Technology
ENZYME PRODUCTION FROM WH
Fermentative production of enzymes
Water Hyacinth biomass is sun-dried, chopped and pulverized to a particle size of about 1-2mm, and is used as substrate for growth of microorganisms
Mineral salt medium containing necessary salts and nitrogen sources is supplemented with the biomass and sterilized.
Inoculated with spores of the cellulase producing organism (Trichoderma reesei , or Aspergillus niger) and cultivated in a bioreactor.
After growth for about 4-6 days, the culture medium is filtered and processed to recover enzyme
Alternatively, the sterilized biomass is spread on trays and inoculated with spores.
Cultivation is carried out in special sterile rooms/cabinets called koji rooms (Solid State fermentation)
After growth, the enzyme is harvested by extraction with citrate buffer
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SCHEMATIC FOR SUBMERGED FERMENTATION
Centrifuge/FilterCulture supernatantCentrifuge/Filter
Pulverized WH
Water Hyacinth
Chopping
Pulverization
Bioreactor
Medium
AcetoneCellulase
Solid mass
Bio gas
Organic Manure Spore
inoculum
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SCHEMATIC FOR SOLID STATE FERMENTATION
National Institute for Interdisciplinary Science and Technology
Pulverized WH
Water Hyacinth
Chopping
Pulverization
Koji RoomTray Fermenter
Medium
Centrifuge/FilterCulture supernatant
Acetone
Centrifuge/Filter
Cellulase
Solid mass
Bio gas
Organic Manure Spore
inoculum
Fermented WHB
Citrate buffer
OTHER ENZYMES
Phytase : Food and Feed Applications
Tannase : Feed Applications
Beta-glucosidase : Synthesis, Bio-ethanol
National Institute for Interdisciplinary Science and Technology
ETHANOL FROM WATER HYACINTH BIOMASS
Ethanol from lignocellulosic biomass is projected as a potential transportation fuel which is renewable
Ethanol is produced by microbial fermentation of sugars
Both hexose sugars (glucose) and pentose sugars (xylose) can be fermented to ethanol by appropriate organisms (Saccharomyces cerevisiae , Pichia stipitis)
Water hyacinth contains cellulose (polymer of glucose) and hemicellulose (rich in xylose) both of which can yield sugars that can be fermented to alcohol
National Institute for Interdisciplinary Science and Technology
ETHANOL FROM WATER HYACINTH BIOMASS: SCHEMATIC
Pulverized WHB
Cellulases
Xylanases
Acid
Pentose sugarsHexose sugars
Fermentation with pentose fermenting microbes (eg Pichia stipitis)
Fermentation with hexose fermenting microbes (eg Saccharomyces cerevisiae)
Fermentation with C5 and C6 co- fermenting microbes (eg recombninant E coli)
Ethanol
Absolute Alcohol
Distillation and Dehydration
National Institute for Interdisciplinary Science and Technology
HEMICELLULOSE UTILIZATION
Water hyacinth is rich in hemicellulose (40-50%)
Any value addition to the water hyacinth should consider the utilization of hemicellulose sugars derived from the biomass
National Institute for Interdisciplinary Science and Technology
Xylitol : a natural food sweetener, a dental caries reducer, and a sugar substitute for diabetics
Can be produced from fermentation of xylose using special yeasts (Candida guillermondii, Candida tropicalis, Debaryomyces hansenii)
2,3-Butanediol: Solvent, liquid fuel, and as a precursor of many synthetic polymers and resins.
Dehydration of 2,3-BD yields the industrial solvent methyl ethyl ketone,which is much more suited as a fuel because of its much lower boiling point. Further dehydration yields 1,3- butanediene, which is the starting material for synthetic rubber and is also an important monomer in the polymer industry
Can be produced by fermentation of xylose using Bacillus polymyxa, Aerobacter aerogenes, Serratia marcescens
Lactic acid : Used in the food, pharmaceutical, and cosmetic industries. It is a component of biodegradable plastic polylactate, the market for which is expected to grow significantly
Can be produced by fermentation of xylose using Lactobacilli
VALUE ADDED PRODUCTS FROM WH-HEMICELLULOSE
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TECHNICAL CHALLENGES
Large Scale Harvesting and Logistics of WHB is the major problem in utilization of this resource for any application including biogasification, ethanol or enzyme production, or even composting
The biomass contains almost 95% of water and the density is as low as 0.3 g/cm3
All methods of collection and pretreatment utilize high quality energy and economy of the processes are poor whether it is gasification or production of enzyme/ethanol
All methods of utilization can only add value to a program where the primary objective is remediation
Positive economic balances can be achieved only through an integrated utilization plan working on a bio-refinery concept
National Institute for Interdisciplinary Science and Technology
BIOREFINERIES FOR WATER HYACINTH PROCESSING
Direct Physical utilization plans with minimal processing
Crafts , Furniture
Composting
Biogas
Needing elaborate processing
Value added compounds
Enzymes
Ethanol
Chemicals
Ethanol, Chemicals
Chemicals
Ethanol
Biogas, Compost Energy
Biogas, Compost Energy
Biogas, Compost Energy
Primary Product
Primary Byproducts
Secondary Byproducts
National Institute for Interdisciplinary Science and Technology
DE-CENTRALIZED UTILIZATION PLANS
Large Scale Plants for bio-processing may not be feasible in certain cases owing to geographical location, large capital investment, sustainable availability of raw material
Centralized Plants are capital intensive, needs more elaborate infrastructure, are technically intensive, and needs skilled manpower
Decentralized operations can be less capital intensive, needs less infrastructure and are less intensive on technical skills
They can generate local employment, income
Financial incentives to end benefactor of remediation programs can attract more people for utilization plans (Collection centres for WH biomass?)
Can we think of co-operatives with public involvement?
National Institute for Interdisciplinary Science and Technology