1_Fun_Bio.pptx
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B.Tech. IV (CH), Semester – VII L T P CES-1: Fundamentals of Biochemical Engineering 3 0 0 3(Elective Subject from Department)• INTRODUCTION (06 Hours)Scope and possibilities; Characteristics and classification of biological matter; Basics of microbial growth; • OVERVIEW OF BIOSEPARATIONS (02 Hours)An Overview of bioseparations; Cell disruptions; Genetically modified organism • SEPARATION METHODS (25 Hours)Filtration; Centrifugation; Adsorption; Extraction; Membrane separation processes; Concepts of precipitation, Chromatography – Basic concepts; Gel filtration Ion exchange chromatography; Hydrophobic chromatography; Affinity chromatography; Suitable examples; Electrokinetic methods of separations; Finishing operations and formulations• INDUSTRIAL APPLICATIONS (12 Hours)Biomass to Biofuels; Bioremediation; Biocatalysts; Biofouling; Microbial Polymer and plastics; Natural resources recovery
Total Contact Hours: 45)BOOKS RECOMMENDED:1.Shuichi Aiba, Arthur E. Humphrey, Nancy F. Millis, “Biochemical Engineering”, 2nd Ed., Academic Press, New York, 1973.2. James E. Bailey, David F. Ollis, “Biochemical Engineering Fundamentals”, 2nd Ed., McGraw hill, 1986.3. L. Weatherley, “Engineering Processes for Bioseparations”, Butterworth-Heinemann Ltd., Oxford, 1995.4. D.L. Pyle, “Separation for biotechnology”, Royal Society of Chemistry, Cambridge, 19945. A. Scragg, “Environmental Biotechnology”, 2nd Ed., Oxford University Press, 2005.
Fundamentals of Biochemical Engineering
Topics already Discussed• The importance of the need of the day of the subject• Similarity and difference between the Chemical and
Biochemical Engineering• Advantages of bioprocesses (ambient temperature,
high product specificity, relatively clean technology)• Application areas like Pharmaceuticals, drug
intermediates, food chemicals, beverages, organic fine chemicals and solvents, industrial enzymes, dairy products etc.
Need for Downstream Processing
• Downstream processing (bioseparation)is essential part of bioprocess Technology
• The products are manufactured using a variety of equipment like Fermenters or bioreactors, other reactors such as airlift, membrane and immobilized cell reactors are also used.
• The products formed are usually in low concentrations and for separation unit operations are involved
Characteristics of Fermentation Broths
The characteristics of fermentation broths that influence the downstream processing of biomolecules include:
•The type of microorganisms and their morphological features (size and shape)
• Concentration of cells, products and byproducts
•Physical and rheological characteristics
Morphology of Cells
Figure 1: Morphology of Cell
•Cells size ranges from 1 micro m to 4000 micro m
•The cells and the cell agglomerates exhibit a variety of shapes.
•Bacterial and yeast cells occur mostly as homogeneously suspended particles in the fermentation broth
•Fungi form a network of hyphi thereby increasing the viscosity of the broth
•Cell size decreases capacity of the separation process decreases and cost increases
Concentration of cells, products and byproducts
The concentrations of the biomass and that of the products in the fermentation broth are important in deciding on the choice of the separation process
Physical and Rheological CharacteristicsThe density of the dry biomass would be about 1400 kg/m3; however the density of the fermentation broth is lower , around 1100 kg/m3 as the cells have a high water content of about 70-80%.
The rheological property of fermentation broth is of importance for downstream processing in the case of centrifugation and membrane separations.
Newtonian and Non-newtonian models can describe most of the fermentation broths.
The viscosity of the broth is strongly influenced by the cell concentration as well as cell shape and to a minor extent by the changes in the concentration of the nutrients and other metabolites.
The stages of bioseparations
The bioseparation stages are:1. Product release and pre-treatment: Involves cell disruption and release of intracellular products2. Removal of insoluble's or particulates that is solid- liquid separation: Filtration or centrifugation achieves the removal or separation of particulates 3. Product concentration and recovery: Involves adsorption, extraction, precipitation or membrane separation4.Purification: Involves high resolution techniques like chromatography and affinity separations5. Finishing operations and formulations: Includes drying, crystallization etc.Note: This is purely for academic purpose and there is no rigid stages for bioseparation
Cell wall•The murein layer is about 10-80 nm thick, made of petidoglycan exists in one form or other in almost all the species.• The space below the murien layer called periplasmic space, is about 8 nm thick and often contains enzymes•The inner layer called the plasma or cytoplasmic membrane is about 8 nm is a double layer made of phospholipids and some proteins and metal ions• The cell interior, called the cytoplasm, is an aqueous solution of salts, amino acids and biopolymers including proteins, enzymes, RNA and DNA.It is necessary to rupture the cell to release the protein in the cytoplasm
Cell Disruption
Cells for disruptions
Physical Methods
Thermoly
sis
For
products of stable of heat
shock
Osmotic shoc
k
Cells in pure water (double amt) , cells
swells
and
disrupts
Ultrasonication
Ultrasound waves of frequency .20kHz
ruptures the
cells
Chemical and Enzymatic methods
Alkali treatment
Detergent solubilization
Cell wall permeabilization
Lipid solubilization by organic solvents
Enzyme digestion
Lytic enz yme to the cell
suspension
Mechanical Disruptions
Bead Mill disruption
High prressure
homogenizer