Introducción

Bioreactores

Prof. Pablo Araujo Granda Ph.D.

paaraujo@uce.edu.ec
Bioreactores
Conceptos generales de Ingeniería
.. Para ”un” reactor …

• Configuración del reactor

• Tamaño del reactor

• Condiciones de proceso (dentro reactor)

• Modo de operación del reactor

 Reactor de Tanque Agitado
Figure 13.4
culture.
Typical stirred-tank fermenter for aerobic

• Mecanismo de Agitación

• Energía alta / Unidad volumen

• Bafles (Vórtices)

• Impeler (Fuerza corte - Velocidad)

• 70-80% ocupación

• Espuma (Rompe espuma)

• Relación diámetro/altura

• Presión hidrostática

• Material de construcción (intercambio

calor/ Esterilización)
 Reactor de Columna (Burbuja)
e 13.5 Bubble-column bioreactor. horizontal plates are sometimes installed in tall bubble col-
umns to break up and redistribute coalesced bubbles.
Advantages of bubble columns include low capital cost, lack of
moving parts, and satisfactory heat- and mass-transfer perfor-
• Dispersión de gas (energía baja)
mance. As in stirred vessels, foaming can be a problem
requiring mechanical dispersal or addition of antifoam to the
• Levaduras / Cerveza / Vinagre /
medium.
Bubble-column hydrodynamics and mass-transfer charac-
Tratamiento de agua
teristics depend entirely on the behaviour of the bubbles
released from the sparger. Different flow regimes occur
depending on the gas flow rate, sparger design, column diame-
• Relación diámetro/altura
ter and medium properties such as viscosity. Homogeneousflow
occurs only at low gas flow rates and when bubbles leaving the
sparger are evenly distributed across the column cross-section.
• Coalescencia burbujas (Evitar)
In homogeneous flow, all bubbles rise with the same upward
velocity and there is no backmixing of the gas phase. Liquid
mixing in this flow regime is also limited, arising solely from
• Bajo costo (mejora coeficientes de
entrainment in the wakes of the bubbles. Under normal
operating conditions at higher gas velocities, large chaotic
calor y masa)
circulatory flow cells develop and heterogeneousflow occurs as
illustrated in Figure 13.6. In this regime, bubbles and liquid

• Características de la burbuja
tend to rise up the centre of the column while a corresponding
downflow of liquid occurs near the walls. Liquid circulation
entrains bubbles so that some backmixing of gas occurs.
• Homogéneo = bajo flujo de gas
Liquid mixing time in bubble columns depends on the flow
regime. For heterogeneous flow, the following equation has
been proposed [4] for the upward liquid velocity at the centre
 13 Reactor Engineering 339

AirLift Reactor
Figure 13.7 Airlift reactor configurations.

Gas exhaust Gas exhaust Gas exhaust

~T

o Do ier

Riser Downcomer

{}

r
i
o] ~
C

0
0
./
parger ....~ : ~ : ~ , ~ 1

t
Air Air Air
t
(a) (b) (c)
 Packed Bed Reactor (Empacado)
3.8 Packed-bed reactor with medium recycle.
• “catalizador” inmovilizado
Gas exhaust
Recirculated m e d i u m
• Entrada de medio (superior o
i
000 0
inferior).
OoOo ~1 ["-'ql'--~ Air
0
Ooo
'J
O0 • Fase líquida contínua entre las
d bed o~oOO ~ ~ Stirred
oo
alyst oOO vessel partículas (recirculación)
cles O Oooo / \
00~00
000
Stirrer Sparger • Transferencia de masa
000
0 O0 (fluído/”partícula”)

Pump • La presencia de gases dificulta

operación.
ised cells and enzymes for production ofaspartate and
e 13.9
Reactor Lecho Fluidizado
Fluidised-bed reactor. Figure 13.10 Trickle-bed reactor.

• Tamaño / densidad / flujo

• Constante movimiento
• Gases entrada salida
• Recirculación de líquido
• Coeficientes de transporte de
masa
• Tratamiento de aguas /
cerveza / vinagre
emergency pressure release, are normally located on the
 Trickle Bed Reactor (goteo)
ckle-bed reactor.

• Variación del empacado

• Líquido (arriba) Spray

• No hay capa contínua de

líquido = gases permitidos

• Tratamientos de agua
residual aerobios
 Consideraciones Construcción
• Acero (presión) / vacio / 3 atm positivas

• Pequeños = tapa removible / plana / torriesférica

• Grandes = agujero de hombre

• Visores de nivel de líquido

• Mínimos sensores: pH, Temperatura, oxigeno disuelto

• Puerto de muestreo esterilizable por vapor

• Entradas de medio / antiespumante / acido / base

 Operación Aséptica
• Contaminación = depende del proceso

• Esterilización con vapor (presión / tiempo)

• Juntas soldadas pulidas &/ no grietas / no

estancamientos

• “todo” lo que entra = estéril / filtros aire 0,01 um

• Válvulas con construcción aséptica

 mic
equipment. For effective sterilisation, all air in the vessel and closure; the main drawback is that these components must be
pipe connections must be displaced by steam. The reactor checked regularly for wear to avoid valve failure. To minimise
con
should be free of crevices and stagnant areas where liquid or costs, ball and plug valves are also used in fermenter construc- con
solids can accumulate; polished welded joints are used in pref- tion.
erence to other coupling methods. Small cracks or Top pinch-bar
gaps in Spindle
With stirred reactors, another potential entry point for con-
joints and fine fissures in welds are a haven for microbial tamination is where the stirrer shaft enters the vessel. The gap
usin
contaminants and are avoided in fermenter construction between the rotating stirrer shaft and the fermenter body must cam

Válvulas
whenever possible. After sterilisation, all nutrient medium and be sealed; if the fermenter is operated for long periods, wear at
air entering the fermenter must be sterile. As soon as flow of the seal opens the way for air-borne contaminants. Several types
Flexible sleeve
steam into the fermenter is stopped, sterile air is introduced to of stirrer seal have been developed to prevent contamination.
ope
valv
maintain a slight positive pressure in the vessel and discourage On large fermenters, mechanical seals are commonly used [ 14]. mu
in t
Figure 13.12 Weir-type diaphragm valve in (a) closed and (b) open positions.
Lower pinch-bar of
tho
) ferm
Industrial fermenters are designed for in situ steam sterilisa- slee
tion under pressure. The vessel should have a minimum fro
number of internal structures, ports, nozzles, connections and the
other attachments to ensure that steam reaches all parts of the ing
Diaphragm equipment. For effective sterilisation, all air in the vessel and clo
pipe connections must be displaced by steam. The reactor che
should be free of crevices and stagnant areas where liquid or cos
solids can accumulate; polished welded joints are used in pref- tion
erence to other couplingf methods. Small cracks or gaps in
(a)
joints and fine fissures in welds are(b)a haven for microbial tam
contaminants and are avoided in fermenter construction bet
 Figure 13.13 Pipe and valve connections for aseptic transfer
ofinoculum to a large-scale fermenter. (From A. Parker, 1958,

Inoculación y muestreo
Sterilization of equipment, air and media. In: R. Steel, Ed,
BiochemicalEngineering,pp. 97-121, Heywood, London.)

Steam Sterileair

• Presión positiva de aire t E

~O
A H
@--t>< 9 I

(industrial / piloto)

• “Juego” de válvulas Steam ~

,, G~~.~
F
<~--~
I Inoculum
vessel

B I
trap
• Vapor / Aire estéril
~ Fennenter~

In these devices, one part of the assembly is stationary while the

 Monitoreo y control
• Ambiente “dentro” = Actividad “óptima”
• T, PH, O2, Flujo, revoluciones, velocidad burbujeo
• Monitoreo y control (Desviación valores deseados)
• Manual / Automatic Feedback / Optimización (basada
en modelos)
• Variables críticas de proceso
• Medidas “on-line” y “off-line”
• Afectaciones (burbujas – OD / densidad BM – pH/O2)
mentation industry; others are currently being developed or these measurements are relatively commonplace; deta
are the focus of research into new instrumentation. descriptions can be found elsewhere [ 17-19]. The availab
Despite the importance of fermentation monitoring, of an on-line measurement or its use in the laboratory does
industrially-reliable instruments and sensors capable of rapid, necessarily mean it is applied in commercial-scale process

Monitoreo y control
accurate and direct measurements are not available for many
process variables. For effective control of fermentations based
Owing to the cost of installation and the financial co
quences of instrument failure during fermentation, meas
on measured data, the time taken to complete the measure- ment devices used in industry must meet stringent performa

Table 13.1 Parameters measured or controlled in bioreactors

Physical Chemical Biological

Temperature pH Biomass concentration

Pressure Dissolved 0 2 Enzyme concentration
Reactor weight Dissolved C O 2 Biomass composition
Liquid level Redox potential (such as DNA, RNA,
Foam level Exit gas composition protein, ATP/ADP/AMP,
Agitator speed Conductivity N A D / N A D H levels)
Power consumption Broth composition Viability
Gas flow rate (substrate, product, ion Morphology
Medium flow rate concentrations, etc.)
Culture viscosity
Gas hold-up
eactor E n g i n e e r i n g J S o

Feedback control loop

ure 13.18 Components of a feedback control loop.

I Disturbances

Set
+
~ Error I Variable,e.g.,pH
~_- Controller
point _ Actuator PROCESS v

Measuredvariable Measurement
device
I-

or fluctuations occur in operating conditions or if cell from metabolic activity. One of the simplest control schem
perties and model parameters change with time, the model is a conventional feedback control loop, the basic elements
y become inadequate and estimation accuracy will decline. which are shown in Figure 13.18. A measurement devi
 Control de proceso

• Instrumento de medida (envía señal a….)

• Controlador / compara señal con “set point” /

Desviaciones = error => acciones a tomar

• Controlador = Persona / electrónico / neumático /

dispositivo de computador … (envía señal a….)

• Actuador = ejecuta la acción de control

 Control de proceso

• PID control = Proportional-Integral-Derivative

• Acción de control = proporcional al error / la

integral del error / derivada del error en tiempo

• Monitoreo simultáneo y ajuste de parámetros

• Digitalización de la señal – DDC (Direct Digital

Control)
 Control de proceso

• Variables = cambios con tiempo (Batch Fed-Batch)

• “constantes” = no es una idea de control “smart”

• Rangos de variación “aceptables” = control

programado.

• Entender detalladamente el proceso

• Modelizar el proceso (reactor)

 Modelización de Procesos
• Se requiere modelización del reactor y del proceso – Matemática

• Descubre que variables monitorear en el proceso ”real”

(operativo).

• Modelo = estructura + parámetros

• Parámetros pueden cambiar en función del tiempo.

• Modelo = Relaciones matemáticas “teóricas” + Observaciones

experimentales

• Modelización contínua = Ec. Diferenciales

• Modelización basada en el Individuo (siguiente unidad) .. J