Scribd Logo
Upload

Welcome to Scribd!

  • 38 views

    Immobilization of Enzymes

    Uploaded by

    Devlina Sengupta
    The document describes immobilizing the enzyme alpha amylase in sodium alginate gel beads and quantifying the protein content. Sodium alginate solution was mixed with enzyme extract and dripped into calcium chloride solution to form beads through ionic crosslinking. The beads were then used to quantify encapsulated protein content through a Bradford assay. Immobilizing enzymes increases reaction efficiency by concentrating enzyme molecules at the surface of a support matrix. Sodium alginate is commonly used due to its mild gelling properties, non-toxicity, and ability to form instant gels with calcium ions.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    Immobilization of Enzymes

    Uploaded by

    Devlina Sengupta
    38 views2 pages
    The document describes immobilizing the enzyme alpha amylase in sodium alginate gel beads and quantifying the protein content. Sodium alginate solution was mixed with enzyme extract and dripped into calcium chloride solution to form beads through ionic crosslinking. The beads were then used to quantify encapsulated protein content through a Bradford assay. Immobilizing enzymes increases reaction efficiency by concentrating enzyme molecules at the surface of a support matrix. Sodium alginate is commonly used due to its mild gelling properties, non-toxicity, and ability to form instant gels with calcium ions.

    Copyright

    © © All Rights Reserved

    Available Formats

    PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    The document describes immobilizing the enzyme alpha amylase in sodium alginate gel beads and quantifying the protein content. Sodium alginate solution was mixed with enzyme extract and dripped into calcium chloride solution to form beads through ionic crosslinking. The beads were then used to quantify encapsulated protein content through a Bradford assay. Immobilizing enzymes increases reaction efficiency by concentrating enzyme molecules at the surface of a support matrix. Sodium alginate is commonly used due to its mild gelling properties, non-toxicity, and ability to form instant gels with calcium ions.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    38 views2 pages

    Immobilization of Enzymes

    Uploaded by

    Devlina Sengupta
    The document describes immobilizing the enzyme alpha amylase in sodium alginate gel beads and quantifying the protein content. Sodium alginate solution was mixed with enzyme extract and dripped into calcium chloride solution to form beads through ionic crosslinking. The beads were then used to quantify encapsulated protein content through a Bradford assay. Immobilizing enzymes increases reaction efficiency by concentrating enzyme molecules at the surface of a support matrix. Sodium alginate is commonly used due to its mild gelling properties, non-toxicity, and ability to form instant gels with calcium ions.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 2

    Immobilization of Enzymes

    Aim:

    To Immobilize alpha amylase in the sodium Alginate Gel and quantify protein content in the
    beads

    Principle:

    We know that enzymes have been used for years in the industry and many other applications;
    so that brings us to the increase for the need of higher qualities of an enzyme. The importance
    of the need for storage and commercialization of this catalytic molecules, has led researcher
    to discover different techniques that can keep them save and stable. Enzyme immobilization
    is one of these techniques, and basically consists in the confinement of an enzyme to a
    matrix/support different from its normal phase, where the absence of substrate and product is
    looked up to. The immobilization increases the number of enzyme molecules per unit area
    increasing the efficiency of the reaction. There are several methods of enzyme
    immobilization like adsorption, covalent binding, cross-linking, encapsulation, membrane
    confinement. In practice, encapsulation is the most preferred method. Common materials
    used in encapsulation are: alginate, chitosan, collagen, carrageenan, pectin & gelatin, silica
    and nanoparticles.
    Alginates, however, are one of the most frequently used polymers due to their mild gelling
    properties and non-toxicity. Alginate is a hydrocolloid, used as a matrix because of its ability
    to absorb water, easy handling and safety, and due to its gelling properties, stabilizers and
    thickeners. Sodium alginate is the most used because of its high solubility in cold water and
    characteristic sol-gel transition in an instant and irreversible way to the calcium ion. In this
    experiment we carry out the spherification process with sodium alginate dissolved in calcium
    chloride to immobilize the fermentation product alpha amylase enzyme.

    Materials Required:
     3% W/V Sodium Alginate solution
     0.2 M Calcium Chloride solution
     Enzyme extract
     3.5 pH acetate buffer (for protein quantification)
     Bradford reagent
     Distilled water
     Glasswares
     Falcon tubes
     5ml Syringe

    Procedure:
     3g of Sodium alginate was dissolved in warm distilled water (not boiling) till all the
    lumps were dissolved. The mixture was allowed to cool.
     To this 100ml of enzyme extract was added and mixed homogenously.
     4.45g of Calcium chloride was weighed and dissolved in 200ml distilled water to
    prepare a solution of 0.2 M concentration. It was stored in a falcon tube at 4 ̊C.
     In a 5ml syringe, the alginate + enzyme extract mix was loaded.
     The mix was slowly dripped through the needle into the 0.2M calcium chloride
    solution. Approximately 45 minutes of gelification time was allowed.
     After gelification was observed, with the help of a strainer and sterile water, the beads
    were filtered.
     The beads were preserved in distilled water in a falcon tube and refrigerated for
    further quantification of their protein content.

    Procedure for quantification of bead protein content:


     6-9 spheres of immobilized beads were taken in a 1.5ml microfuge tube.
     To this 500μl of pH 3.5 acetate buffer was added and the beads were grounded
    thoroughly using a glass rod.
     This was dissolved in 500μl distilled water and vortexed thoroughly.
     Total protein content was quantified by Bradford assay from a standard curve
    prepared by taking few standard protein concentrations and their respective
    absorbances.

    You might also like