Documentos de Académico
Documentos de Profesional
Documentos de Cultura
Biotechnology
Ideal situation
Population
Industry
Farming
Waste
Atomsferic systems
Terrestrial systems
Aquatic systems
In reality
Large accumulation of WASTE in
dump sites, soil and water systems
Environmental
Biotechnology
Application of biological agents, along with
chemical, physical and engineering processes
to maintain, protect or restore environment
Environmental
Biotechnology
Biomass utilization (waste and by-product)
Treatment of liquid and solid waste from
domestic
municipal
agricultural
industrial sources
Bioremediation (removal of toxic waste
from the environment)
Biomass utilization
Biomass:
source material for production of economically important products
material of biological origin, excluding biological fossils like coal,
natural gas or oil
by-products of the food and agricultural industries (discarded in
the past) (=waste)
e.g. starch, lignocellulose, whey, meat flour, part of municipal
waste, activated sluge, etc.
Biofuel
Bioethanol and biobutanol
Biodiesel
Biomethanol
Biohydrogen
EU Directive for Biofuel (2003/30/CE) set goals for energy
obtained from waste (not food) for the transportation sector:
2005 2%
2010 5,75%
2020 10%
Biofuel
Additives/substitutes to oil derivatives
Bioethanol
may be produced directly from sugar-rich plants (sugar cane, sugar beet)
starches from foodstuffs (corn, wheat, sorghum, grain) require a hydrolysis reaction
(saccharification), to convert the starch into sugars, either enzymatically (amylases) or
chemically
lignocelluloses and hemicelluloses (straw, wood) usually are subjected to chemical
hydrolysis
hydrolyzed wastes (household, other) may also be used
Bioethanol
Sacharomyces cerevisiae
Zymomonas mobilis
high specific productivity (g ethanol g-1 cell h-1)
high ethanol yield (g ethanol g-1 substrate)
high alcohol tolerance
narrow substrate range, difficult GE
Improving tolerance of the fermentation micro-organisms to ethanol
Improving the stoichiometric yield of ethanol per molecule of sugar
Improving the ability of the organism to utilise lignocellulose
Lignocellulose waste
Very abundant and non-edible, but very challenging
Cellulose: composed of glucose residues; 33 % of plant biomass
Hemicellulose: xylose, mannose, galactose, rhamnose, and arabinose
- sugars not readily fermented; 20 % of plant biomass
Lignin: phenylpropane polymer; 30 % of all non-fossil organic carbon
Engineering
xylose
utilization
for ethanol
production
Heterologous
enzymes
Overexpression
Biobutanol
Clostridium acetobutylicum + starch
Acetone, butanol and ethanol (3:6:1), hydrogen and other by-products
Higher energy density
Ease of blending with conventional fuels
Higher toxicity of butanol to the producing organism
Production from sugar is three fold less efficient
than for ethanol production
Chemical and thermal routes for production are dependent on
a clean source of syngas (CO + H2) from biomass gasification
Acetone-Butanol
Fermentation
Enzymes are indicated by letters as follows:
(A) glyceraldehyde 3-phosphate dehydrogenase
(B) pyruvate-ferredoxin oxidoreductase
(C) NADH-ferredoxin oxidoreductase
(D) NADPH-ferredoxin oxidoreductase
(E)NADH rubredoxin oxidoreductase
(F) Hydrogenase
(G) phosphate acetyltransferase (phosphotransacetylase)
(H) acetate kinase
(i) Thiolase (acetyl-CoA acetyltransferase)
(J) 3-hydroxybutyryl-CoA dehydrogenase
(K) Crotonase
(L) butyryl-CoA dehydrogenase
(M) phosphate butyltransferase (phosphotransbutyrylase)
(N) butyrate kinase
(0) acetaldehyde dehydrogenase
(P) ethanol dehydrogenase
(Q) butyraldehyde dehydrogenase
(R) butanol dehydrogenase
(S) acetoacetyl-CoA:acetate/butyrate:CoA transferase
(T) acetoacetate decarboxylase
(U) phosphoglucomutase
(V) ADP-glucose pyrophosphorylase
(W) granulose (glycogen) synthase
(X) granulose phosphorylase
Micribiologcal Reviews 1986, 50: 484-524
Algae to fuels
Algal fuel factories can absorb waste carbon dioxide emissions from
coal power plants or other polluting entities
Certain algae in a high carbon dioxide environment will produce a viable
fuel similar to diesel
Biodiesel
First engine invented run on
groundnut oil (Rudolf Diesel)
Diesel, fraction of crude oil composed
mainly of saturated hydrocarbons
Use of vegetable oils for making biodiesel, less polluting and renewable nature
Biodiesel mixture of methyl esters, which is made from the transesterification
of plant oils
Modern diesel engine can not run solely on vegetable oil due to high viscosity,
deposit formation on injection systems and poor cold-start properties
World biodiesel production (2006) about 5-6 million tonnes, with 4.9 million
tonnes processed in Europe (Germany, Italy, Austria, France and Sweden)
Biodiesel is currently limited to 5% in diesel in Europe due to concerns over
engine warranties, materials, cold weather performance and compatibility
Germany is the major biodiesel producer (2499 x106 L, 2006)
Biodiesel
Feedstock derived from renewable sources, mainly vegetable plant oil
Type and yield of fatty acids accumulated vary on crop, but all contain triacylglycerols
Soybean (USA and South America), rapeseed (Europe), sunflower, palm, coconut,
peanut, corn, castor, cottonseed, radish, rice bran, hemp, canola, sorghum
Jatropha, jojoba
Algae
Wastes, as recycled cooking oil, waste animal grease
Oleaginous microorganisms (including genetically modified)
Fatty acid
Soybean
Sunflower
Palm
Rapeseed
Palmitic (C16:0)
11
42.8
Stearic (C18:0)
4.5
Oleic (C18:1)
23
18
40.5
11
Linoleic (C18:2)
54
69
10.1
12
Linolenic (C18:3)
0.2
Erucic (C22:1)
52
Biodiesel
Transesterification - reduces the viscosity of the oil, improves its consistency and miscibility
with diesel, as well as improving other properties, such as its viscosity when cold.
Fatty acids reacts with low molecular weigh alcohol, usually methanol, under acid or
alkaline catalysis (0.5% NaOH or KOH)
Enzymes can also be used as catalyst
Supercritical methanol technology without catalyst
The ester of the fatty acid is formed together with glycerol (10:1, w/w)
Energetics of Biofuel
Pathways to fuel
Plsticos
Polmeros mais usados
Polipropileno - PP
Petrleo
Totalmente
NO
BIODEGRADVEIS
Poliestireno - PS
Problemas ambientais
Adaptado de: Vtor Alves, CEER Centro de Engenharia dos Biossistemas ISA/UTL
90 m
Polmeros
biodegradveis
Bioplsticos
tais
como
amido,
Podem ou no ter
origem em
fontes
renovveis
protenas e celulose
Biodegrabilidade
Decomposio em dixido de carbono, gua, metano, compostos
inorgnicos ou biomassa, por aco enzimtica de
microorganismos presentes no ambiente, num perodo de tempo
definido.
A biodegrabilidade no est directamente relacionada com a origem das matriasprimas, mas sim com a natureza qumica do polmero e com as propriedades finais
da embalagem/produto
Existem polmeros derivados do petrleo que so biodegradveis
Polmeros Biodegradveis
Biomassa
vegetal/animal/algas
Polissacridos
Amido
Protenas
Hemicelulose
Alginato
Carragenato
Pectina
Quitosano
Monmeros da
petroqumica
Polihidroxialcanoatos
Poli(cido lctico)
(PHAs)
(PLA)
Policaprolactona
(PCL)
Polissacridos
De soja
(batata)
Celulose
Monmeros
microbianos
De soro de leite
(milho)
(trigo)
Origem
microbiana
Polieteramidas
(PEA)
(gelano)
Gelatina
(xantano)
(bovinos)
(peixe)
(celulose
bacteriana)
Glten
(alginato
bacteriano)
(pululano)
Polisters alifticos
(p.e. PBSA)
Polisters aromticos
(p.e. PBAT)
Polihidroxialcanoatos (PHAs)
Produzidos por bactrias
Acumulados dentro das clulas em grnulos
Reservas de energia
Fontes de carbono: acares, cidos orgnicos volteis,
glicerol (de preferncia
presentes em resduos e subprodutos)
Poliester
Propriedades semelhantes
ao polipropileno
Hidrofbico
Barreira a CO2 e O2
Barreira vapor de gua
Lactic Acid
Lactic acid bacteria
Lactobacillus, Sporolactobacillus, Enterococcus, Lactococcus,
Bacillus, Streptococcus, Pediococcus, Leuconostoc
Two different pathways of lactic fermentation exist:
homolactic - lactic acid is the only product
Heterolactic produces lactic acid plus various co-products such as ethanol, carbon
dioxide and acetate
Bifidobacteria
They have a unique hexose metabolism
(catabolize sugars via frutose-6-phosphate phosphoketolase),
Are exclusively anaerobic (others are generally microaerophilic)
Lactic Acid
Used
as a preservative and iron chelator in food
in cosmetics
in pharmaceuticals
Produced by
Lactic acid bacteria, that require:
complex media, pH control
Unsuccessful!
Abbott, DA. FEMS Yeast Res 9 (2009) 11231136
Alkanes (- oil)
Candida tropicalis
Candida oleophila
Saccharomycopsis lipolytica
Alkanes are insoluble, but cells produce emulsifying substances and are enriched in
lipids in order to consume droplets by passive diffusion
Methane
(not all is used as fuel)
Obligate methylotrophs
Methylomonas methanica
Methylococcus capsulatus
Methylovibrio soehngenii
Methanomonas margaritae
CH4
+ O2 + XH2
HCHO
Methanol
(inhibitor)
+ H2O
Methanol
Bacteria:
Rapid growth
Higher protein content
Better yields
Simpler culture medium requirements
Catabolism of:
Methanol
Methyamine
Formaldeyde
Formate
CH3OH
HCHO
HCOOH
CO2
One-carbon
assimilation pathways
Serine Pathway
Serine pathway
formaldehyde
Quayle cycle
Transketolase and transaldolase
Wood-Ljungdahl pathway
CO2
Calvin Cycle (ribulose diphosphate, plants)
Quayle Cycle
EMP
ED pathway
2 ATP
2 NADH
1 ATP
1 NADH, 1 NADPH
One-carbon
assimilation pathways
Quayle cycle
(Ribulose monophosphate cycle):
most efficient, selected for use
WasteWaste-water treatment
Increased need due to:
Increase in world population
Agriculture dependence on fertilizers and pesticides
Man-made chemicals
Necessary due to:
The need to reclaim wastewater
Preventing contamination of drinking water
Preventing entry in the food chain
Should treat (amongst others):
Compounds with high organic load (BOC)
Pathogenic organisms and viruses (cholera, hepatitis, thyphoid,
parathyphoid)
Man-made chemicals
Sewage, industrial effluents, agricultural run-off, storm water and urban runoff
ESTRATGIA GERAL
lquidos
no aquosos
solutos
perigosos
correces
fsicas (temperatura)
qumicas (pH, bio-nutrientes)
EFLUENTE
AQUOSO
elevao
tanque de
igualizao
agentes
slidos/lquidos precipitantes /
floculantes
flutuantes
colector
municipal
elevao
slidos
suspensos
grosseiros
densos
tanque de
igualizao
resduo slido
TRATAMENTO PRINCIPAL EM
ETAR PRPRIA /
PARQUE INDUSTRIAL
DESCARGA
REUTILIZAO
PRELIMINAR /
PRIMRIO
slidos
suspensos
leves
TERCIRIO
SECUNDRIO (AFINAO)
CBO
turbidez
(carncia
poluentes diludos
bioq. de O2) patogneos
N, P
CAUDAIS A TRATAR
UNIDADES INDUSTRIAIS
Balanos mssicos
Higiene de equipamentos e instalaes
gua da rede potvel
??? %
GUAS RESIDUAIS
guas pluviais
MUNICPIOS
infiltrao
consumos de gua
domstico
industrial
(na malha urbana)
UTILIZAO
60-85%
GUAS RESIDUAIS
pblico
rede de saneamento
no contabilizado
(perdas; erros de medida)
Gilda Carvalho, IBET
+
rede pluvial
(sarjetas)
rede
unitria
Tratamento de
guas residuais
ETAR Terras de Santa Maria
Matria Orgnica
Carncia Qumica de Oxignio (CQO) mede o teor de matria orgnica presente numa gua
residual, por oxidao qumica, a quente, em meio cido.
Agente oxidante utilizado: Dicromato de potssio, K2Cr2O7
Agente acidificante: H2SO4 concentrado
Temperatura da digesto qumica: 160C
Tempo da digesto qumica: 110 min
Carncia Bioqumica de Oxignio CBO5(20) permite medir a matria orgnica presente
numa gua residual, atravs de oxidao biolgica, a 20C, durante cinco dias
Agente oxidante utilizado: microrganismos presentes na gua residual, ou uma populao
de microrganismos (inculo) previamente adaptada gua residual
Temperatura da digesto biolgica (incubao): 20C
Tempo da digesto biolgica (incubao): 5 dias
Luminosidade: no escuro, para reduzir a actividade de produo de oxignio pelos
organismos fotossintticos que possam estar presentes na gua residual
No Mtodo Respiromtrico, ao longo dos 5 dias de incubao, procede-se determinao
do oxignio consumido ao fim de cada perodo de 24 horas
Agitao: permanente para facilitar as trocas gasosas entre a amostra e a atmosfera
existente no interior do Respirmetro
e ainda slidos totais, slidos totais em suspenso, C total, C orgnico total, carncia
total de oxignio
Organic Matter
Secondary treatment
(biological, with
activated sluge microbial consortia):
Decrease BOD
Aggregation
Adsorption
(Polysaccharides,
metallothioneins)
Biological treatment
Aerobic (activated sluge)
surface aeration
OR
trickling filter
(with sand and stones)
Anaerobic digestion
on sediment sludge from aerobic process
occasionally on raw sewage
TRATAMENTO AERBICO
- LAMAS ACTIVADAS
Consrcio microbiano
ADSORO
CBO
CBO
CBO + O2
NUTRIENTES
azotados
fosfatados
CBO residual
solvel
coloidal
suspenso
O2
CO2 + H2O
NO3 -
NO2
O2
NO3 -
O2
OUTROS
(p. ex. nutrientes
residuais)
bactrias, fungos
BIOMASSA
(separvel da
gua tratada)
Denitrification
(anoxic or anaerobic)
O2
O2
Remoo de Fsforo de
guas Residuais
Tratamento Qumico
remoo: precipitao com sais de ferro ou alumnio (cloreto frrico, almen)
pros: eficiente, de baixo custo de investimento e relativamente simples
cons: elevados custos (produtos qumicos utilizados, volume acrescido de
lamas), maior contedo em sais no efluente final; quantidade de
precipitante qumico aumenta exponencialmente na razo inversa da
quantidade de fosfato a obter no efluente
Tratamento Biolgico
remoo: crescimento microbiano (consrcio); acumulao intracelular
(polifosfato)
pros: efluente com muito baixa concentrao de fsforo, sem custos
adicionais, maior flexibilidade (conjugada com a remoo biolgica de
azoto); lamas utilizadas como fertelizante aps estabilizao
cons: eficincia e estabilidade do processo (reversibilidade)
Anaerobiose/Aerobiose
Efluente
REACTOR
ANAEROBIO
Efluente
(baixa [P])
REACTOR
AEROBIO
Purga
Biomassa
Recircula o de lamas
Anaer
Aerob
PHA
Glicognio
Crescim.
Carbono
Fosforo
Tempo
Methane Production
The main goal is not always energy production, but stabilization of sludge
Only a small fraction of the production is distributed to others
Methane is insoluble thus readily removed from the reaction mixture
but
Its long-distance transport is difficult
1 kg of sludge 600 l of methane (60-70% of produced gas, inc. CO2 and H2S)
There is a risk of explosion
Methane Production
Methane production: final link in the anaerobe food chain
Microbial consortia are required
Starting material: cellulose, starch, fats and protein - methanogenics dont use!
Methane production depends on anaerobic fermentative bacteria:
Polymer hydrolysis facultative anaerobes
Production of low chain fatty acids (sludge liquefaction phase) other anaerobes
Direct substrates for methanogenesis:
Acetate (75 %), methanol, formate, H2 + CO2 (almost all the rest)
Yield depends on substrate composition and temperature.
Improvement: Multistage process
Temperature optimization (45-60C)
Mixing
Methane Production
Syntrophomonas
Syntrophobacter
Methanobacterium
thermoautotrophicum
Methanosarcina
Composting
Useful for:
Soil conditioning material
Commercial production of
Agaricus bisporus
Providing rich microflora and
fauna for bioremediation
Composting
Aerobic solid state fermentation
Controlled oxidation of organic matter to
produce a stable and humified product
(it may be necessary to add a N-source in order to obtain a C/N of 30:1)
Microbial consortium depends on substrate composition and temperature
(commercial inoculants also for sale):
1.
2.
3.
4.
5.
Composting
Solid waste
Bioremediation
Removal of toxic waste
from the environment
using biological agents
Bioremediation
Technology used to speed up the natural processes of waste degradation and
recycling
Use of (naturally occurring) microorganisms such as bacteria, fungi, and yeast to
degrade pollutants or hazardous substances in soil, water and air into non-toxic
or less toxic substances
Biologically-based remediation detoxifies hazardous substances instead of
merely transferring contaminants from one environmental medium to another
Bioremediation is generally less disruptive to the environment than excavationbased processes
The cost of treating a hazardous waste site using bioremediation technologies
can be considerably lower than that for conventional treatment methods:
vacuuming, absorbing, burning, dispersing, or moving the material
Bioremedia\on biodeterioration
Xenbiotico
Biodegradvel
Sofre degradao primria, parcial ou mineralizao
Persistente
Degradao muito lenta em certo ambiente
Recalcitrante
No est biodisponvel ou no sofre degradao em
certo ambiente
Recalcitrant
compounds
Hydrofobic nature
Biomagnification or bioconcentration
(organisms higher up the food chain)
Xenobiotics
Bio-degradable
petroleum products (gas, diesel, fuel oil) crude oil compounds (benzene, toluene,
xylene, naphthalene) some pesticides (malathion) some industrial solvents coal
compounds (phenols, cyanide in coal tars and coke waste)
Partially degradable
TCE (trichlorethylene) threat to ground water PCE (perchlorethlene) dry cleaning
solvent PCBs (have been degraded in labs, but not in field work) Arsenic,
Chromium, Selenium
Not degradable / Recalcitrant
Uranium Mercury DDT
In natural communities
Halogenated compounds, especially halogenated aromatic compounds (such as
polychlorinated biphenyls) are often recalcitrant
The presence of halogens in a meta position makes the compound more recalcitrant
Often one stereoisomer of an organic compound will be biodegradable, while another
isomer will be recalcitrant
Specific organisms in an environment may be able to degrade recalcitrant compounds,
at varying rates depending on the conditions
Degradation of xenobiotics
Xenobiotic as sole energy source
may lead to
Mineralisation: conversion of organic to inorganic compounds
(opposite of partial degradation)
Co-metabolised with another substrate (usually when microbial
enzymes synthesized for the degradation of another substrate also
degrade the xenobiotic)
may lead to
Primary biodegradation: disappearance of parent compound
Intermediates can be more toxic and persistent than parent compound
(activation), or may become accessible to further degradation
Physiological conditions
Requirements for biodegradation:
Electron acceptor at a residual level of 1 ppm or more
Essential organic and inorganic nutrients
Microbes and substrate must be in contact
Water
Oxidation of xenobiotics
Faster oxidation
when O2 is the
electron acceptor
Fe3+
HCO3SO42-
NO3-
Alvarez, Pedro J. J., Illman, Walter A., Bioremediation and Natural Attenuation, 2006
Degradation enzymes
are plasmid-encoded
Plasmid-encoded
genes
Biostimulation
Biodegradation by naturally-occurring organisms may be stimulated by
- adding essential nutrients to the contaminated area
- providing aeration or limiting aeration, depending on whether the
contamination is better degraded under aerobic or anaerobic conditions
- using plants and the microbial communities of their rhizospheres
(phytoremediation)
Bioaugmentation
Adding microbes not normally found in an environment to try to alter or accelerate
the biodegradation process
When the microbes are added without consideration of their normal habitat (e.g.,
just adding a pure culture), there may be short-term improvement but the added
microbe usually fails to establish a stable population
Better results are may be seen when the added organisms microenvironment
(nutrients, oxygen, aeration, etc.) are included in the bioaugmentation strategy
Metal bioremediation
Remediation of heavy metals such as:
lead, chromium, arsenic, and others
in soil and groundwater
frequently uses reduction reactions to lower the solubility of an
oxidized metal (lowers two or three valence states)
Ultimately, the metal is precipitated as a sulfide or in another
immobile and low toxicity form
It may also be retained in the biomass bioabsortion
A soil washing technique may also be coupled to metal reduction
Radioactive bioremediation
Uranium
One of the heaviest elements found in nature, 1.7 times denser than lead
emits alpha, beta, and gamma particles
The half-life of U238 is 4.5 billion years!
By-products (such as Thorium (Th232)) have even longer half-lives,
making uranium an element that gets more radioactive during its own
decay process
Natural uranium is 99.274% U238, 0.720% U235, and 0.0055% U234
Uranium is enriched when its content of U235 (fissible) reaches 3.2-3.6%
Radioactive bioremediation
Radioactive bioremediation
Uranium contamination:
is a threat to wildlife and water supplies
can cause kidney damage and cancer in humans
takes billions of years to decay radioactively
traditional clean-up methods involving pumping of contaminated water
are useless against uranium because it sticks to soil
Radioactive bioremediation
Sequestring and/or reducing metals
Geobacter metallireducens
reduces uranium
Shewanella oneidensis
produces UO2 with extracellular cyochrome
Deinococcus radiodurans
radiation resistent
Geobacter species have been shown to play important roles in the bioremediation of
groundwater contaminated with petroleum and landfill leachate. Soluble organic
contaminants are oxidized to carbon dioxide with the reduction of iron oxides that are
abundant in most subsurface environments. Molecular analysis of a diversity of
contaminated subsurface environments in which oxidation of pollutants coupled to iron
oxide reduction is important have consistently found that these environments are
enriched in Geobacter species
Petroleum
In addition to pollution through human activities, millions of tons of
petroleum enter the marine environment every year from natural seepages.
Despite its toxicity, a considerable fraction of petroleum oil entering marine
systems is eliminated by the hydrocarbon-degrading activities of microbial
communities, in particular the so-called hydrocarbonoclastic bacteria.
Alcanivorax borkumensis
n-alkane degradation
biosurfactant production
biofilm formation
scavenging of nutrients and cofactors in the oligotrophic saline marine
environment
copes with various habitat-specific stresses
Legislation
Many strains not used due to concerns with:
horizontal gene transfer
persistence of strain in the environment
Bioleaching
Thiobacillus, Bacillus, Pseudomonas
Thermofilic: Thiobacillus, Sulfolobus
Many other candidates!
Thiobacillus ferrooxidans and thiooxidans
Chemoautotrophs
Electron donnors: Fe2+ (to Fe3+), molecular and reduced sulfur (to sulphate)
Its important to provide physiological growth conditions
Bioleaching
Copper
2 S + 3 O2 + 2 H2O
2 CuFeS2 + H2SO4 + 8.5 O2
CuS + 2 O2
2 H2SO4
2 Cu(SO4) + Fe2(SO4)3 +H2O
Cu(SO4)
Fe2(SO4)3 + H2SO4
Fe2(SO4)3 + FeS2
FeSO4 + 2S
Uranium
hexavalent U extracted with hot H2SO4 and Fe3+
UO2 + Fe2(SO4)3
Bioleaching
Field Applications
Laboratory
Recycling cells
Recycling leachate
Use of finely ground ores
Shaking
Temperature control
Good yields
Recovery by
precipitation
organic solvent addition
ion-exchange
microbial methods
Crueger & Crueger 1894. Biotechnology