INTRODUCTION Lecturer Dr Kamal E M Elkahlout Assistant P rof of Biotechnology Relationship of Scientists Engineers Microbiologists biochemists and molecular biologists are scientists welltrained in empirical testing of hypotheses ID: 436210
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Advanced Bioprocess EngineeringINTRODUCTION
Lecturer Dr. Kamal E. M. ElkahloutAssistant Prof. of BiotechnologySlide2
Relationship of Scientists Engineers
Microbiologists, biochemists, and molecular biologists are scientists, well-trained in empirical testing of hypotheses.Engineers develop theories based on mathematical models, use models to predict performance, optimize and develop processes.Slide3
Biologists and Engineers
Research scientists often pursue knowledge while applications may take a secondary role.The work of engineers is often driven by economics of an application and problem solving.Slide4
A bioprocess
is any process that uses complete living cells or their components (e.g., bacteria, enzymes
,
chloroplasts
) to obtain desired products
Transport of energy and mass is fundamental to many biological and environmental processes.
Areas
, from
food processing
to thermal design of building to
biomedical devices
to
pollution control
and
global warming
, require knowledge of how energy and mass can be transported through materials[
mass,momentum,heat
transfer]Slide5
Bioprocess Engineering
It is a specialization of Biotechnology,
Chemical Engineering
or of
Agricultural Engineering
.
It
deals with the design and development of equipment and processes for the manufacturing of products such as
food
,
feed
,
pharmaceuticals
,
nutraceuticals
,
chemicals
, and
polymers
and
paper
from biological materials.
Bioprocees
engineering is a conglomerate of mathematics, biology and industrial design
,Slide6
It consists of various spectrums like designing of
Fermentors, study of fermentors (mode of operations etc). It also deals with studying various biotechnological processes used in industries for large scale production of biological product for optimization of yield in the end product and the quality of end product.
Bio process engineering may include the work of mechanical, electrical, and industrial engineers to apply principles of their disciplines to processes based on using living cells or sub component of such cells.Slide7
Definition of Fermentation
Metabolism: energy generating processes where organic compound acts as both electron donor and acceptor.Industrial Biotechnology: the process by which large quantities of cells are grown under aerobic or anaerobic conditions.The industrial microorganisms are grown under controlled conditions with an aim of optimizing the growth of the organism for production of a target microbial product.Slide8
Definition of Fermentation
Fermentation is carried out in vessels known as FermentorsThe types of fermentor ranges from simple tank to complex integrated system of automated control.Slide9
An overview of a typical industrial fermentation process and the movement of
materials through a typical fermentation plant are shown in the following figure:
Fermentation PlantSlide10
The Process of Fermentation
Process is divided into a number of sections:· In-bound logistics: (the delivery and storage of raw materials) · Upstream processing: the processing of raw materials for the fermentation
· The fermentation, where the major conversion occurs
· Downstream processing: the purification and concentrating of the raw product(s)
· Out-bound logistics : the final packaging, storage and delivery of the purified product(s)
Stages of Industrial fermentation:
· Upstream Processing (USP)
· Downstream Processing (DSP)Slide11
Upstream Processing
The upstream part of a bioprocess refers to the first step in which microbes/cells are grown, eg bacterial or mammalian cell lines (see Cell culture), in bioreactors
.
B
asically
up
stream
processing involve all those steps related with
inoculum
development, media development, improvement of
inoculum
by genetic engineering process, optimization of growth kinetics so that product development can improve tremendously. Slide12
Fermentation has two part upstream & downstream. A
fter product development the next step is purification of product for desired quality. When they reach the desired density (for batch and fed batch cultures) they are harvested and moved to the downstream section of the bioprocesSlide13
Upstream Processing
Three main areas: A) Producer microorganism
This include processes for
obtaining a suitable microorganism
strain improvement to increase the productivity and yield
maintenance of strain purity
preparation of suitable
inocullum
B ) Fermentation media
C) Fermentation ProcessSlide14
Downstream Processing
The downstream part of a bioprocess refers to the part where the cell mass from the upstream are processed to meet purity and quality requirements. Downstream processing is usually divided into three main sections,
a
capture section,
a
purification section and
a
polishing section. Slide15
The volatile products can be separated by distillation of the harvested culture without pre-treatment.
Distillation is done at reduced pressure at continuous stills. At reduced pressure distillation of product directly from fermentor may be possible. Slide16
The steps of downstream processing are:
1-separation of biomass 2-cell disruption
3-concentration of broth
4-initial purification of metabolites
5-metabolite specific purification
6-de-watering
7-polishing of metabolitesSlide17
1-separation of biomass: separating the biomass (microbial cells) generally carried out by centrifugation or ultra-centrifugation. If the product is biomass, then it is recovered for processing and spent medium is discarded. If the product is extra cellular the biomass will be
discarded.Ultra filtration is an alternative to the centrifugation.2-cell disruption:
If the desired product is intra cellular the cell biomass can be disrupted so that the product should be released. The solid-liquid is separated by centrifugation or filtration and cell debris are discarded.
Slide18
3-concentration of broth: The spent medium is concentrated if the product is extracellular.
4-initial purification of metabolites: According to the physico-chemical nature of the product molecule several methods for recovery of product from the clarified fermented broth were used ( precipitation, solvent extraction, ultra-centrifugation, ion-exchange chromatography, adsorption and solvent extraction)5-metabolite specific purification:
specific purification methods are used when the desired metabolite is purified to a very high degree.Slide19
6-de-watering: If low amount of product is found in very large volume of spent medium, the volume is reduced by removing water to concentrate the product. It is done by vacuum drying or reverse osmosis.
7-polishing of metabolites: this is the final step of making the product to 98 to 100% pure.the purified product is mixed with several inert ingredients called EXCIPIENTS. the formulated product is packed and sent to the market for the consumers.
Slide20Slide21Slide22
Types of Fermentation Process
Batch FermentationContinuous FermentationFed batchBatch reactors ,simplest type. Reactor is filled with medium and the fermentation is allowed.
Fermentation has finished, contents are emptied for downstream processing.
The reactor is then cleaned, re-filled, re-inoculated and the fermentation process starts again.Slide23
Types of Fermentation Process
Continuous reactors, where fresh media is continuously added and bioreactor fluid is continuously removed. As a result, cells continuously receive fresh medium and products and waste products and cells are continuously removed for processing.The reactor can thus be operated for long periods of time without having to be shut down.
Many times more productive than batch reactors.Slide24
Types of Fermentation Process
does not have to be shut down as regularly the growth rate of the bacteria in the reactor can be more easily controlled and optimizedcells can also be immobilized in continuous reactors, to prevent their removal.Fed batch reactor, most common type of reactor used in industry.
fresh media is continuous or sometimes periodically added
.Slide25
Batch Fermentation Process
dynamic processes that are never in a steady state. Often , the critical parameter is gas exchange or balance between respiration rate and oxygen transfer. sterilized media components are supplied at the beginning of the fermentation with no additional feed after inoculation.Slide26
Batch Fermentation Process
cells are grown in a batch reactor, they go through a series of stages:Lag phase· Exponential phase· Stationary phase· Death phaseSlide27
Batch Fermentation ProcessSlide28
Batch Fermentation Process
Lag Phase microbial population remains constant as there is no growth. However it is the period of intense metabolic activity.Factors Influencing the Lag Phase
·
Chemical composition
of the fermentation media influences the length of the lag phase.
Longer lag phase is observed if the
inocullum
is transferred into a fresh medium of different carbon source.
· Age of the
inocullum
. If the
inocullum
is in exponential growth phase, it will exhibit shorter lag in the fresh medium.
·
Concentration of the
inocullum
.
·
Viability and morphology of the
inocullum
.Slide29
Penicillin: “Birth of Biochemical Engineering”
1928- Alexander Flemming was plating Staphylococcus aureus and the plate was contaminated with mold – near the mold no bacteria grew. WWII- most common cause of death was infection from wounds.Sulfa drugs were effective on limited range of infectious organisms.1930-1940 British scientists Florey and Chain at Oxford developed a process to produce penicillin from the mold.Slide30
How Penicillin Works...
Antibiotics on a plate: cell walls do not formSlide31
Early Work
They asked US pharmaceutical companies to help work on the project – to develop a commercial scale process for penicillium.Merck, Pfizer, Squibb, USDAAt this time, most drugs were made synthetically. Fermentation was unproved and companies were skeptical.Problem: low concentrations, fragile product.Slide32
Significant Advances
New medium- Corn steep liquor (x10).New strain isolated from molded fruit- P. chrysogenum (still used in some form).Change to tanks from “bottle plants”.Separation: liquid-liquid extraction.Slide33
Challenges
Very large (10 kgal) fermentation vessels.Provide sterile air and feed.Agitator seal.Heat removal.Recovery and purification of fragile product.Slide34
Biology-Engineering ConnectionCooperation between engineers and scientists was critical (Merck specifically formed teams of each).
“Biochemical engineering” born as a result.Slide35
• Essential Knowledge/
Topics needed in Bioprocess engineering work– Cells– How cells work– Metabolism Pathways– Enzymes and Kinetics
– Reactors: Design and Data Analysis
–
Bioseparations
– Application and Industrial considerationsSlide36Slide37Slide38Slide39Slide40Slide41Slide42Slide43
Cleaning in place/sterilization in place
Oxygen utilization rate
Oxygen transfer rateSlide44Slide45Slide46Slide47Slide48Slide49Slide50Slide51Slide52Slide53Slide54