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    Introduction To Downstream Processing

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    AkshayaaRavee
    Downstream processing refers to the recovery and purification of products such as pharmaceuticals from natural sources like fermentation broth. It involves removing insolubles, isolating the desired product from other components, and purifying it through separation of close contaminants. The final processing steps polish the product to a stable, transportable form for packaging. Proteins have diverse functions related to their complex hierarchical structures determined by amino acid sequences and various bonding interactions.

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    Introduction To Downstream Processing

    Uploaded by

    AkshayaaRavee
    30 views24 pages
    Downstream processing refers to the recovery and purification of products such as pharmaceuticals from natural sources like fermentation broth. It involves removing insolubles, isolating the desired product from other components, and purifying it through separation of close contaminants. The final processing steps polish the product to a stable, transportable form for packaging. Proteins have diverse functions related to their complex hierarchical structures determined by amino acid sequences and various bonding interactions.

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    Lecture 1 DP Introduction

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    Downstream processing refers to the recovery and purification of products such as pharmaceuticals from natural sources like fermentation broth. It involves removing insolubles, isolating the desired product from other components, and purifying it through separation of close contaminants. The final processing steps polish the product to a stable, transportable form for packaging. Proteins have diverse functions related to their complex hierarchical structures determined by amino acid sequences and various bonding interactions.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    30 views24 pages

    Introduction To Downstream Processing

    Uploaded by

    AkshayaaRavee
    Downstream processing refers to the recovery and purification of products such as pharmaceuticals from natural sources like fermentation broth. It involves removing insolubles, isolating the desired product from other components, and purifying it through separation of close contaminants. The final processing steps polish the product to a stable, transportable form for packaging. Proteins have diverse functions related to their complex hierarchical structures determined by amino acid sequences and various bonding interactions.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 24

    Introduction to Downstream Processing

    Downstream processing refers to the recovery and purification of biosynthetic


    products, particularly pharmaceuticals, from natural sources such as animal or plant
    tissue or fermentation broth

    Removal of insolubles
    involves the capture of the product as a solute in a particulate-free liquid

    Product Isolation
    is the removal of those components whose properties vary markedly from that of
    the desired product.
    Product Purification
    is done to separate those contaminants that resemble the product very closely in
    physical and chemical properties.
    Product Polishing
    describes the final processing steps which end with packaging of the product in
    a form that is stable, easily transportable and convenient.
    Down stream Processing of Proteins

    Enzymology
    Proteins are the center of action in biological processes.
    Nearly all the molecular transformations that define cellular
    metabolisms are mediated by protein catalysis
    One of the keys to deciphering the function of a given protein is
    to understand its structure.
    By examining how the amino acids are strung together, we can
    endeavor to understand the chemical and physical properties of
    proteins and, ultimately, their mechanism of action in living
    organisms.
    Functions
    • Diverse functions related to structure
    – Structural components of cells
    – Motor proteins
    – Enzymes
    – Antibodies
    – Hormones
    – Hemoglobin/myoglobin
    – Transport proteins in blood
    Structure

    • Amino acids
    – Amino group
    (NH2)
    – Carboxyl group From: Elliott, WH. Elliott, DC. (1997) Biochemistry
    and Molecular Biology. Oxford: Oxford University
    (COOH) Press. p23
    Amino acids

    • 20 amino acids make up


    protein
    • 8 essential amino acids
    – 9 in infant (histidine)
    From: Elliott, WH. Elliott, DC. (1997) Biochemistry and
    • Dipolar
    Molecular Biology. Oxford: Oxford University Press.
    p23
    – +ve end (NH3+)
    – -ve end (COO-)
    Protein structure - bonding

    • 5 bonds or forces determine structure


    – Peptide bond
    – Hydrogen bond
    – Disulfide bond
    – Ionic bond
    – Hydrophobic force
    Peptide bond

    • Peptide bond joins


    amino acids
    • Bond at both ends
    – Increases range of
    possible proteins

    From: Elliott, WH. Elliott, DC. (1997) Biochemistry


    and Molecular Biology. Oxford: Oxford University
    Press. p23
    Primary protein structure Primary structure of insulin

    • Linear sequence of amino


    acids forms primary
    structure
    • Sequence essential for
    proper physiological
    function

    Bettelheim & March (1990) Introduction


    to Organic & Biochemistry
    (International Edition) Philadelphia:
    Saunders College Publishing, p299
    Sickle cell anemia

    • Replacement of
    single glutamine
    with valine in one
    polypeptide chain
    of hemoglobin
    alters structure and
    function Bettelheim & March (1990) Introduction to Organic & Biochemistry
    (International Edition) Philadelphia: Saunders College Publishing, p301
    Secondary protein structure

    • Peptide chains fold into secondary


    structures:
    –  - helix
    –  - pleated sheet
    – Random coil
     - helix

    • Shape maintained
    by hydrogen
    bonds between
    C=O and N-H
    groups in
    backbone
    • R groups directed
    outward from coil
    From: Elliott, WH. Elliott, DC. (1997) Biochemistry and Molecular
    Biology. Oxford: Oxford University Press. p28
     - pleated sheet

    • Structure
    maintained by
    hydrogen bonds
    between C=O and
    N-H groups in
    backbone
    • R groups directed
    above and below
    backbone From: Elliott, WH. Elliott, DC. (1997) Biochemistry and Molecular
    Biology. Oxford: Oxford University Press. p29
    Random coil
    • Not really random
    structure, just non-
    repeating
    – ‘Random’ coil has fixed
    structure within a given
    protein
    – Commonly called
    ‘connecting loop region’
    From: Elliott, WH. Elliott, DC. (1997) Biochemistry – Structure determined by
    and Molecular Biology. Oxford: Oxford University
    Press. p27 bonding of side chains
    (i.e. not necessarily
    hydrogen bonds)
    Torsion angles of the polypeptide backbone

    The Ramachandran plot


    Tertiary protein structure

    • Secondary structures fold and pack together


    to form tertiary structure
    – Usually globular shape

    • Tertiary structure stabilised by bonds


    between R groups (i.e. sidechains)
    Tertiary structure - H bond

    • H bonds weak
    allowing to be
    broken and reformed
    Hydrogen bond
    easily
    – Allows structural
    change
    • produces ‘functional’
    molecules
    Tertiary structure - disulfide bond

    – Covalent bond
    between sulfur
    atoms on two
    cysteine amino acids

    From: Elliott, WH. Elliott, DC. (1997) Biochemistry


    and Molecular Biology. Oxford: Oxford University
    Press. p32
    Tertiary structure - ionic bond

    • Ions on R
    groups form salt
    bridges through
    ionic bonds

    From: Summerlin, LR. (1981) Chemistry for the Life Sciences. New
    York: Random House, p459
    Tertiary structure - hydrophobic forces

    • Close attraction of non-polar


    R groups through dispersion
    forces
    • Very weak but collective
    interactions over large area
    stabilise structure
    • Repel polar and charged
    molecules/particles Bettelheim & March (1990) Introduction to Organic & Biochemistry
    (International Edition) Philadelphia: Saunders College Publishing, p302
    Quaternary protein structure

    • Arrangement of multiple
    tertiary structures into
    single functional
    complex
    • Allows for changes in
    structure/function in
    response to chemical
    stimuli From: Elliott, WH. Elliott, DC. (1997) Biochemistry and
    Molecular Biology. Oxford: Oxford University Press. p27

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