5/3/2012

Polticas de Desarrollo de Gobierno : Tecnologa

Academia Chilena de Ciencia: redefini las polticas
de investigacin
Sectores que contribuyen al desarrollo:
1. Gestin Econmica
2. Hortofruticultura
3. Biotecnologa
4.Biohidrometalurgia
5. Salmonicultura
6. Nutricin
7. Educacin y Generacin de softwares y redes
informticas

5/3/2012
El crecimiento tradicional del negocio
minero se logra a travs de dos rutas:
Actividades que mediante una fusin o adquisicin
de los activos de una operacin, pueden alcanzar un
crecimiento en el corto plazo.
Tradicional exploracin, el cual demanda gran
inversin de dinero y tiempo y que basan su real xito
en el hallazgo y que la industria denomina
elefante. Alcanza un crecimiento a muy largo plazo.

5/3/2012
Existe un enfoque nuevo y alternativo
Sin embargo, su xito radica en que la
tecnologa sea de competencia nica:
Biotecnologa.
Tecnologa Palanca: Capaz de entregar oportunidades
de crecimiento en el mediano plazo, si la tecnologa es
la apropiada.

5/3/2012
La explosin de las ciencias biolgicas abre
nuevas perspectivas de desarrrollo a las
fronteras de las ciencias de la tierra
Las biotecnologas ofrecen perspectivas para la
identificacin de nuevos microorganismos
adaptados a condiciones extremas
Pueden ser utilizados para el desarrollo de nuevos
procesos (ecoprocesos) :
- tratamiento de minerales, rocas, suelos, napas y
deshechos
- fluidos geotrmicos, formaciones geolgicas para
diversos fines: explotacin,depuracin,
almacenamiento, etc
El ciclo de la materia orgnica y del CO2 en lo
procesos geolgicos


5/3/2012
BIOTECNOLOGIA
Es el uso integrado de ciencias de la ingeniera,
microbiologa y bioqumica que alcanzan su
aplicacin tecnolgica (industrial) mediante el
aprovechamiento de las capacidades que poseen los
microorganismos, sus cultivos o partes de ella.
Incluye:
- Manipulacin y produccin de microbios
- Purificacin y separacin de productos biolgicos
para el tratamiento de slidos y lquidos.
Aplicacin qumica, farmacetica, mdica,
alimentos, agricultura e industria minera.

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Biotecnologa Mineral
Biomining : Disolucin microbial de valores
metlicos a partir de minerales
Biobeneficio : Remocin microbial selectiva
de constituyentes indeseables
Funcin de los microorganismos : cataliza la
disolucin o genera productos metablicos
que causan la disolucin.
Uso directo de microorganismos y sus
derivados en el procesamiento mineral y en
la bioremediacin de deshechos de la
industria minera
5/3/2012
Biomining
Es una forma de procesamiento mineral que
utiliza microorganismos para degradar
sulfuros metlicos para obtener mejores
recuperaciones de metales con valor
econmico
Se ha desarrollado como uno de las ms
exitosas reas de la biotecnologa, con un
valor global del proceso de 10 billones de
US$ (1999)
5/3/2012
Interaccin Microbio-Mineral
Genera un proceso en presencia microbial,
el que puede intervenir mediante
fenmenos fsico-bioqumicos :
- Alteracin de la superficie qumica de
slidos
- Generacin de superficie qumicamente
activas
- Disolucin selectiva de fases slidas de una
matriz
- Sorcin,acumulacin y precipitacin de iones
y compuestos
5/3/2012
Microbe-Mineral Interaction
Bio-beneficiation can be defined as a
process brought about by microorganism
which mediate in a number of surface-
chemical and physico-chemical phenomena
such as:
- Alteration of the surface chemistry of
minerals
- Generation of surface-active chemicals
- Selective dissolution of mineral phases in
ore matrix
- Sorption,accumulation and precipitation of
ions and compounds
5/3/2012
Introduction
Activities of mining explotation
cause ores resources of low grade,
principally as complex sulphides.
Most used extractive process is
flotation-fusion-converting.
Its generates dangerous levels of
environmental contamination, unless
incorporate expensive clean
mechanisms
5/3/2012
Biohydrometallurgical processes
include
- bioleaching: microbial dissolution of
metal values.
- biobeneficiation: selective microbial
removal of undesirable consituents
Both processes exploit
biogeochemical activities of
microorganism
5/3/2012

Microorganism are increasingly finding
use in mineral processing and
hydrometallurgy both for the
enhancement of mineral enginering
operations and for the remediation of
mineral industry wastes




5/3/2012
Some of the applications, such as biologically
assisted leaching of sulfide ores and biooxidation
of refractory sulfides gold ores are established
commercial processes.
Others, such as the use of organisms for the
removal of heavy metals ions from dilute aqueous
streams
Use of microorganisms in leaching non-sulfide
ores, the flocculation or flotation on minerals and
remediation of toxic chemical discharged from
mineral engineering operations.
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SCOPE
Successful commertialization of bio-
oxidation for refractory gold and for low
grade copper, and the potential for its use
in a wide number of mineral processing and
environmental control operations
there is considerable incentive to gain
further understanding of :
- Identification of the sub-processes
involved
- their mechanism and kinetics,
in order to identify the rate limiting
sub-processes.
5/3/2012
Two basically different large
scale processes can be applied in
bacterial leaching:
Nowadays in-situ dump or heap
l}eaching are the most widespread
bacterial leaching processes.
Aerated slury reactors find less
application:
- investment cost of slurry reactors
- Production costs determined by
aeration, heat transfer , pyrite
oxidation rate.
- Improvement of the oxidation rate
per unit of reactor volume is essential
for the feasibility.
5/3/2012
ANTECEDENTES MICROBIOLOGICOS
Ingeniera Bioqumica
Procesamiento de material biolgico
Utilizacin de agentes biolgicos
Teora Celular (Schleiden y Schwann, 1840)
Un sistema vivo se compone de un mdulo bsico
CELULA
Anlisis de los seres vivos
Estudio individual de la clula
Estudio completo del organismo

Seres Vivos
Reino Animal Reino Vegetal Reino
Protisto
Protistos : Organismo que no presenta un
desarrollo de tejidos y pueden ser uni o
pluricelulares microorganismos

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Caractersticas
Autalimentacin o Nutricin : Toma
sustancias qumicas del ambiente la
transforma, libera energa y elimina
productos de deshecho
Autoduplicacin o desarrollo: Dirige
su propia sntesis
Interaccin a travs de seales
qumicas
Evolucin: Cambios hereditarios que
permiten seleccionar a los
microorganismos mejor capacitados
para vivir en determinados ambientes.

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P Pr ro oc ca ar ri io ot te es s E Eu uc ca ar ri io ot ta as s
O Or rg ga an ni iz za ac ci i n n c ce el l u ul la ar r Unicelular Pluricelular
P Pa ar re ed d Rgida (cido
diamlico pimlico
D.A.P)
No posee rigidez
R Re es sp pi ir ra ac ci i n n Aerbica o anaerbica Aerbica
D Di iv vi is si i n n No Mitsica Mitosis
A Ap pa ar ra at to o n nu uc cl le ea ar r Sin membrana
nuclear
Con membrana
nuclear
CLASIFICACION CELULAR
Observaciones SEM revela dos tipos
marcadamente diferentes de clulas
tanto en funcin y organizacin
Difieren en la forma de resolver
problemas comunes de la vida celular


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CELULA : C , H, O, N, P y S
Estructuras celulares
Monmeros: bloque individual de
construccin
Macromolculas: monmeros
conectados en forma especfica
Tipos de Monmeros
Azcares : constituyente monmero
de POLISACARIDOS
Acidos grasos: constituyente
monmero de LIPIDOS
Nucleticos : constituyente monmero
de ACIDO NUCLEICO
Aminocidos : constituyente
monmero de PROTEINA

Polmeros No-informativos: Secuencia
repititiva de monmero y su secuencia
carece de importancia funcional.Lpido
y Poli
Polmeros Informativos:Secuencia
monmera muy especfica que
contiene informacin biolgica.
Protenas
ENZIMA
Protena catalizadora de reacciones
metablicas es altamente especfica :
cada enzima cataliza un solo tipo de
reaccin.
Catlisis enzimtica : 108 a 109 veces
la velocidad espontnea
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CAPACIDADES
METABOLICAS
Metabolismo : Serie colectiva de
procesos qumicos que se llevan a cabo
en organismos vivos.
Nutrientes: Sustancia qumica del
medio ambiente, a partir de los cuales
se forma una clula.

Transformaciones Qumicas en una
Clula
1. Anabolismo: Una clula se construye
a partir de nutrientes: Es un proceso
de BIOSINTESIS donde se requiere
energa.
2. Catabolismo: Sustancias qumicas
usadas como fuente energtica, se
rompen en compuestos ms sencillos y
lo hacen liberando energa.
DEGRADACION

Metabolismo es funcin de:
1. Fuente energtica
2. Fuente de carbn
3. Aceptor o donor de
electrones

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CLASIFICACION DE
ACUERDO A CAPACIDADES
METABOLICAS
1. Basado en la fuente energtica para
crecer y biosntesis

a. Fottrofa : Utilizan la luz solar.
b. Quimitrofa : Utilizan la energa
qumica

2. Basado en la fuente de carbn

a. Auttrofa : Utilizan dixido de
carbono
b. Hetertrofa : Usan compuestos
diferentes al CO2

3. De acuerdo a la fuente de e- que
usan para sus Rx OX-RED

a. Organtrofa : Si el donante de e-
es un compuesto orgnico.
b. Littrofa : Si el donante de e- es
un compuesto inorgnico.
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1 1. . A Ae er r b bi ic co os s : : Existen en presencia
de oxgeno Respiracin
O O
2 2
2 2. . A An na ae er r b bi ic co os s: : Como aceptor de e-,
utilizan compuestos inorgnicos
oxidados.
2.1 Desnitrificacin
2.2 Reduccin de sulfato
2.3 Reduccin de Fe3+
2.4 Reduccin de protones
2.5 Reduccin de S
N NO O
3 3
- -
S SO O
4 4
2 2- -
F Fe e
3 3+ +
H H
+ +
S S
3 3. . F Fe er rm me en nt ta ac ci i n n : : Los compuestos
orgnicos sirven dsimultneamente
de donores y aceptores de e-
MODOS METABOLICOS
De acuerdo a la naturaleza del
oxidante y del reductor

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Bioenergtica
Estudio cuantitativo de las
transformaciones de energa
Naturaleza y funcin de los procesos
qumicos en las que se basan estas
transformaciones.
- Basadas en leyes termodinmicas
- Adenosintrifosfato (ATP):
biomolcula de alta energa que
participa en el flujo de energa celular
La energa qumica se conserva
gracias a la formacin de ATP
acoplada con las reacciones
exergnicas degradatorias del
catabolismo
La energa qumica se utiliza luego, por
rompimiento del ATP, para las
reacciones sintticas endergnicas
del anabolismo y para otros procesos,
ejm. cambio de propiedades de
protenas
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Bioenergtica
ATP es el portador universal de
energa metablica y es el nexo entre
el catabolismo y al anabolismo
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Bioenergtica
Crecimiento de A. Ferrooxidans es el
fenmeno bioenergtico ms inusual.
Caractersticas del estilo de vida de
A. Ferrooxidans:
Pequea cantidad disponible de
energa disponible en el sustrato para
el crecimiento
Bacteria quimilittrofa requiere fijar
su propio CO2 y si es necesario su
propio N2.
Es obligadamente acidfila
Membrana celular posee un contenido
altamente especfico de cadena
transportadora de electrones

5/3/2012
Crecimiento Bacterial
t0 X0
t1 X1= 2Xo Primera generacin
t2 X2= 2X1 = 22X0 Segunda
generacin
t3 X3= 2X2 = 23X0 Tercera
generacin
t4 X4= 2X3 = 24X0 Cuarta
generacin

tn Xn= 2Xn-1 = 2nX0 n-sima
generacin

ln Xn = n ln2 + ln X0
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Cultivos batch y continuos
Medicin de flujo de gases
Cultivo
Conjunto de microorganismos a los que se
les ha hecho crecer deliberadamente en un
medio adecuado a escala de laboratorio





Tcnicas Experimentales Estudio de la
Cintica





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Medio de cultvo
Una solucin nutriente : 9K
Un sustrato adecuado: en solucin o slido



Composicin g / l
(NH4)2SO4 3
KCl 0,1
K2HPO4 0,05
MgSO4 .7H2O 0,5
Ca(NO3)2.4H2O 0,0215

Concentracin ptima de Fe 2+ : 10 g/l

5/3/2012
BIO-OXIDACION DEL ION FERROSO
Oxidacin de Fe
2+
:
Fe3+ + e- = Fe2+
Reduccin del O2 :
2 H2O = O2 + 4 H+ + 4 e
________________________________
4 Fe2+ + O2 + 4 H+ = 4 Fe3+ + 2 H2O Eh
= 0,77 + 0,059 log (Fe3+ / Fe2+)


5/3/2012

5/3/2012
Bacteria
Thiobacillus ferroxidans,
Leptospirillum ferrooxidans and
Thiobacillus thiooxidans are obligate
chemolithoautotrophic bacteria :
- Use chemical energy.
- Energy from inorganic substrate.
- Their sole or major source of
carbon for cell biosynthesis is carbon
dioxide.
T. ferrooxidans : ferrous iron,
numerous sulphides, sulfur,
thiosulphate and polythionates
L. Ferrooxidans : ferous iron
T. Thiooxidans: sulphur and its
inorganic compounds
5/3/2012
DIRECT MECHANISM IN BIO-
OXIDATION OF METAL
SULPHIDES
It is assumed that irreversibly
attached bacteria to the mineral
surface and the sulphur moiety is
biologically oxidized.
Specific mechanism, enzymatic or
electrochemical, by which this occurs
has not yet been elucidated.

5/3/2012
Direct Mechanism

5/3/2012
INDIRECT MECHANISM IN
BIO-OXIDATION OF METAL
SULPHIDES
It is assumed that the sulphide
mineral is chemically oxidized by
ferric iron leading to the dissolution
of the metal cation.
Chemical oxidation reaction can be:
- complete ( ferrous iron and sulphate
are formed) and the role of the
bacteria is to regenerate ferric iron
by oxidizing the ferrous iron.
- incomplete (ferrous iron and
elemental sulphur are produced) the
rol of bacteria is to oxidize the
sulphur to sulphate.
5/3/2012
Methods used in Literature to
discriminate between the direct
and indirect mechanism
1. Scanning electron micrographs
Some author observed the formation
of pits and corrosion patterns in the
bacterial oxidation and concluded
that direct bacterial attack of the
surface took place. However, pit
Fornmation was also found to occur
in chemical oxidation of mineral.
SEM shown substantial intergranular
variations in surface texture and
these grain edges and corners,
defectcs, solid and fluid inclusion pit,
cleaveages and fractures were both
inherited from mineral growth
history.This sides were chemically
reactive because of high excess
surface energy and this explained
the ocurrence of pit formation.
5/3/2012
4. Bio-oxidation at iron free conditions
Some authors claimed a direct mechanism bycomparing a bio-
oxidation rate that was greater than the chemical, sterile leaching
rate at equal initial concentrations of ferric iron. However this
conclusion is incorrect, because in the sterile blank the used ferric
iron was not regenerated and ferrous iron accumulated..
5/3/2012
INDIRECT MECHANISM IN BIO-OXIDATION OF METAL
SULPHIDES
It is assumed that the sulphide mineral is chemically oxidized by
ferric iron leading to the dissolution of the metal cation.
Chemical oxidation reaction can be:
- complete ( ferrous iron and sulphate are formed) and the role of
the bacteria is to regenerate ferric iron by oxidizing the ferrous
iron.
- incomplete (ferrous iron and elemental sulphur are produced)
the rol of bacteria is to oxidize the sulphur to sulphate.
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Indirect Mechanism
5/3/2012
Prelimnary
Update, the kinetics equation most used is that Pinches:


This has been shown to fit both batch and continuou data and to
predicts multi-stages pilot plant perfomance.
Ftting continuous steady state data is successful, but the major
limitation is that it is not mechanistically based.

|
|
.
|

\
|
=
m
m
X
X
1 X k
t d
X d
5/3/2012
Methods used in Literature to discriminate between the
direct and indirect mechanism
1. Scanning electron micrographs
Some author observed the formation of pits and corrosion patterns in the
bacterial oxidation and concluded that direct bacterial attack of the surface
took place. However, pit Fornmation was also found to occur in chemical
oxidation of mineral.
SEM shown substantial intergranular variations in surface texture and these
grain edges and corners, defectcs, solid and fluid inclusion pit, cleaveages
and fractures were both inherited from mineral growth history.This sides
were chemically reactive because of high excess surface energy and this
explained the ocurrence of pit formation.
5/3/2012
3. Biomass yield on the mineral sulphide
Some authors claim that based on a comparison of the bacterial
growth yields on ferrous iron and the metal sulphide, a direct
mechanism has to be postulated.
Arkesteyn showed that the transport of electrons from sulphur to
oxygen yielded more energy for the bacterial CO
2
fixation than the
transport of electrons from ferrous iron.
5/3/2012
4. Bio-oxidation at iron free conditions
Some authors claimed a direct mechanism bycomparing a bio-
oxidation rate that was greater than the chemical, sterile leaching
rate at equal initial concentrations of ferric iron. However this
conclusion is incorrect, because in the sterile blank the used ferric
iron was not regenerated and ferrous iron accumulated..
5/3/2012
2. Cyclic Voltammetry

Technique of studying the kinetics of elevtrode processes : current
from the electrode is monitored as the potential of elctrode is
changed.
Measured cyclic measurements of pyrite electrodes in presence
and absence of T. Ferrooxidans were closely matched.
Slight differences were attributed to the presence of small amounts
of iron transferred with the inoculum into the experimental
solution.
5/3/2012
Engineers need to have a picture on whether a direct bacterial
oxudation of sulphide minerals is possible in principle, and secondly
on whether this is s relevant (rate determining) sub-processes with
respect to the overall bio-oxidation rate of sulphide minerals.
Determination of a reaction mechanism by analytical technique,
is often recognized as a scientifically solid method. However, from
literature, it has not proven to be conclusive with respect to the
bio-oxidation mechanism.
5/3/2012
THEORICAL AND EXPERIMENTAL METHODS IN
DETERMINING RATE-LIMITING SUB-PROCESSES
Chemical and bacterial oxidation rates presented in the literature
were mostly determined from off-line concentration
measurements in batch experiments.
Although this is an accepted method, it is limited in the accurate
determination of the oxidation rates as a function of the process
conditions, and consequently, in the determination of rate limiting
sub-processes.
5/3/2012
It is shown how general theory and experimental methods
used in other fields of the biotechnology can be applied to
kinetics experiments in biohydrometallurgy.
In the literature, often cell counts, organic nitrogen or protein
analyses were applied to determine the biomass concentration .
However, these methods are time consuming, and suffer of limited
accuracy when used to determine the bacterial growth rate.
Mass balance have not been checked in any of the kinetics studies.
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Mass and Charges Balances
Assuming that the biomass composition is represented by CH
1,8
O
0,5

N
0,2
and introducing Y
sx
as the biomass yield on ferrous iron, the
following stoichiometric equation for the bacterial oxidation of ferrous
iron results:
O
2
H f
3
Fe e
0. 2
N
0. 5
O
1. 8
CH
H d
2
Fe
sx
Y
1
2
O c
4
NH b
2
CO a
+
+
+

+
+
+
+ +
+
+
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Conservation of elements and charge must be provided
O
2
H
sx
Y 2
sx
1,2Y 1
3
Fe
sx
Y
1
0. 2
N
0. 5
O
1.8
CH
H
sx
Y
sx
0,2Y 1
2
Fe
sx
Y
1
2
O
sx
Y 4
sx
4. 2Y 1
4
NH 0. 2
2
CO
|
|
.
|

\
|
|
|
.
|

\
|
|
|
.
|

\
|
|
|
.
|

\
|

+
+
+
+

+
+
+

+
+
+
5/3/2012
Relation between the oxidation rate of ferrous iron and the
oxygen and carbon dioxide consumption rates: Degree of
Reduction Balance
2 2
2
O CO
Fe
r 4 r 4,2 r - =
+
This equation shows the effect of using an integrated
stoichiometric equation for ferrous iron oxidation and biomass
growth compared with :

2
2
O
Fe
r 4 r - =
+
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Yield and Maintenance
Primary process in baterai oxidation reactions is considered to be the
oxidation of substrate. These oxidation reactions generate energy that
is used by bacteria for growth and maintenance.
According the empirical Pirt relationship, the following equation
predicts the substrate oxidation rate that is required for both the
growth and maintenance:

x
C
s
m
Y
x
r
s
r
max
sx
+ =
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Specific Rates
Biomass specific oxidation rate of compound i, is defined as the rate
per C-mole of biomass:

q i= ri /Cx
So, the oxygen consumption rate per C-mole of biomass:
qO2 = r O2 /Cx
Biomass specific growth rate, , is defined by:
= rx /Cx


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s
m
max
sx
Y
1
sx
Y
1
+ =
Using the definitions of actual yield of biomass on substrate and
specific growth rate, Pirts relationship can be rewritten:

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Similary, introducing the maximun yield of biomass on
oxygen and the maintenance coefficient of biomass on
oxygen, the oxygen consumption rate required for growth
and maintenance of bacteria on a certain substrate
according to the Pirt equation is:

o
m
max
ox
Y
1
ox
Y
1
+ =
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Propositions
On line oxygen and carbon dioxide off-gas analyses
Determination of the biomass concentration from the carbon
dioxide analyses in the off-gas and applying the carbon balance
The ferrous iron ( any substrate) oxidation rate is calculated from
the oxygen and carbon dioxide consumption rate using the degree
of reduction balance
Application of a biological oxygen monitor as an analytical tool to
measure the oxygen consumption rate
of cell suspensions from batch or continuous cultures.
5/3/2012
Methods to Determine Rate Limiting Sub-processes
Simultaneous substrate oxidation experiments are perfomed in
the presence and absence of bacteria at equal ferrous to ferric iron
ratios and pH.
Other bacterial substrates oxidation kinetics are examined by the
measurement of the biomass specific consumption rate of
substrate grown cells in the presence and absence of substrate, at
equal ferrous to ferric iron ratios.
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CONCLUSIONS
Theorcal fundam,entals used in biotechnology can be apply to
identify of sub-processes involved in bio-oxidation
Degree of reduction balance is an integrated stoichiometric
equation.
Through Pirts equation it can determine the energetic parameters
involved on the process
Theorical results can be experimentally verified by measurements
on line of variables

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En 1903, Henry propuso que la enzima se combina con la molcula de sustrato
para formar el complejo enzima-sustrato como paso necesario en la catlisis. L.
Michaelis y M. Menten, en 1913, ampliaron ese fundamento para proponer que la
enzima E, se combina con el sustrato S en forma reversible y rpida para formar el
complejo enzima-sustrato, ES, el que puede disociarse para generar el producto
de reaccin P y liberar la enzima E en un paso ms lento, el cual limita la
velocidad global, segn(6):
p E
4
k
3
k
ES
2
k
1
k
S E + +




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Al inicio de la reaccin, la concentracin del producto es insignificante de tal
manera que se puede despreciar la constante k
4
, de tal manera que la velocidad
inicial queda determinada por la descomposicin de ES.
Dado que la concentracin de complejo enzima-sustrato es difcil de medir
experimentalmente, su cuantificacin se logra a travs de una expresin analtica:
De este modo, las velocidades de formacin y descomposicin de ES :
sustrato en libre total
E E E + =
| || | S E de formacin de Velocidad
1
k ES =
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