Transcript of Advanced Bioprocess Engineering INTRODUCTION Lecturer Dr. Kamal E. M. Elkahlout Assistant Prof. of...
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- Advanced Bioprocess Engineering INTRODUCTION Lecturer Dr. Kamal
E. M. Elkahlout Assistant Prof. of Biotechnology
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- 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.
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- 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.
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- A bioprocess is any process that uses complete living cells or
their components (e.g., bacteria, enzymes, chloroplasts) to obtain
desired productscellsbacteria enzymeschloroplasts 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]food
processingbiomedical devicespollution controlglobal warming
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- Bioprocess Engineering It is a specialization of Biotechnology,
Chemical Engineering or of Agricultural
Engineering.BiotechnologyChemical EngineeringAgricultural
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.foodfeedpharmaceuticals
nutraceuticalschemicalspolymerspaper Bioprocees engineering is a
conglomerate of mathematics, biology and industrial design,
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- It consists of various spectrums like designing of Fermentors,
study of fermentors (mode of operations etc). Fermentors 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.
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- 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.
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- Definition of Fermentation Fermentation is carried out in
vessels known as Fermentors The types of fermentor ranges from
simple tank to complex integrated system of automated control.
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- 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 Plant
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- 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)
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- 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.Cell
culture bioreactors Basically upstream 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.
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- Fermentation has two part upstream & downstream. After
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 bioprocesdownstream
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- 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 Process
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- 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.upstream
Downstream processing is usually divided into three main sections,
a capture section, a purification section and a polishing
section.
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- 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.
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- 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 metabolites
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- 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.
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- 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.
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- 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.
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- Types of Fermentation Process Batch Fermentation Continuous
Fermentation Fed batch Batch 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.
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- 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.
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- 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 optimized cells 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.
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- 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.
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- Batch Fermentation Process cells are grown in a batch reactor,
they go through a series of stages: Lag phase Exponential phase
Stationary phase Death phase
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- Batch Fermentation Process
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- 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.
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- 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.
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- How Penicillin Works... Antibiotics on a plate: cell walls do
not form
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- Early Work They asked US pharmaceutical companies to help work
on the project to develop a commercial scale process for
penicillium. Merck, Pfizer, Squibb, USDA At this time, most drugs
were made synthetically. Fermentation was unproved and companies
were skeptical. Problem: low concentrations, fragile product.
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- 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.
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- Challenges Very large (10 kgal) fermentation vessels. Provide
sterile air and feed. Agitator seal. Heat removal. Recovery and
purification of fragile product.
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- Biology-Engineering Connection Cooperation between engineers
and scientists was critical (Merck specifically formed teams of
each). Biochemical engineering born as a result.
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- 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 considerations
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- Cleaning in place/sterilization in place Oxygen utilization
rate Oxygen transfer rate
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