Biotechnological enzyme production

Course Coordinator


Course Description


1. Determination of enzyme kinetic parameters for the substrate conversion to product under conditions of enzyme industrial application

L: The history of enzyme technology development. Selection of enzymes for industrial application: determination of kinetic parameters and influence of certain parameters on enzyme reaction rate and process productivity of industrial biocatalysis. Influence of bioprocess parameters on enzyme activity and enzyme stability in conditions of enzyme industrial application. Applied kinetics of enzyme reactions. The influence of enzyme inhibitors and activators in industrial processes.

S: Calculation of the required free/immobilized enzyme for implementation of the biocatalysis in industrial conditions.

2. Biotechnological production of free enzymes

L: Microbial biosynthesis of enzymes: medium composition, bioprocess parameters, inoculum preparation, solid-state and submerged cultivation conditions, bioreactor designs. Enzyme isolation: general scheme, precipitation, extraction, cell disruption, (ultra)filtration and chromatography techniques. Modern, advanced and alternative separation techniques in biopharmaceutical production. Strategies to improve enzyme yield. Product formulations.

S: Biotechnological production of free enzymes for the industrial application.

E: Wohlgemuth`s method. Anson`s method. Biosynthesis of a-amylase with Bacillus subtilis. Purification of active enzyme filtrate. SDS-PAGE electrophoresis of enzyme samples.

3. Biotechnological production of immobilized enzymes

L: Enzymes immobilization and stabilization. Methods of enzyme immobilization for industrial application. Application of hybrids and composite materials as nanoparticles to obtain most effective enzyme immobilisation with increased applicability of biocatalytic systems. Membrane reactors in enzyme technology. Influence of environmental parameters and diffusion constraints on the immobilized enzyme kinetics.

S: Biotechnological production of immobilized enzymes for industrial application.

E: Application of enzyme microencapsulation: Immobilization of a-amylase in agar. Immobilization of alkaline proteases in calcium alginate. Determination of immobilized enzymes activities.

4. Industrial application of enzymes

L: Kinetic comparisons of enzyme bioreactors. A review of certain industrially important enzymes: proteolytic, amylolytic, pectinolytic, cellulose, penicillin-amidase and L-aminoacylase. Application of enzymes in biotechnological, food and other industries, analytical, scientific and use in biosensors. Application of genetically modified microorganisms and enzyme engineering for the production of enzymes with better catalytic properties. Implementation of “omics” technologies to facilitate the discovery of new microbial enzymes for industrial production and application. Application of enzymes from extremophile microorganisms. Application of enzymes in two-phases systems. Development of one-pot multienzyme systems to conduct multistage enzymatic reactions. Legislation regarding the application of industrial enzymes in food production and as food additives.

S: Application of enzymes in biotechnology, food and other industries


  • Select the enzyme for a particular industrial application with the best-fit values of the kinetic parameters of the enzymatic reaction on a large scale
  • Compare enzymes activity and stability, as important parameters regarding industrial applications of enzymes
  • Present schematic diagram of the biotechnological process of enzyme biosynthesis by submerged cultivation of microorganisms and the processes of isolation and purification of enzymes
  • Present the main parameters of biotechnological production of enzymes under solid-state cultivation
  • Select enzyme isolation and purification procedures, depending whether the enzyme is extracellular or intracellular
  • Compare the pros and cons of the classical immobilization methods with standard carriers and nanoencapsulation methods with hybrid and composite materials as enzyme carriers
  • Explain the working principle of membrane bioreactors in enzyme technology
  • Prepare the inoculum, perform biosynthesis of amylolytic enzymes by submerged cultivation of Bacillus subtilis, and isolation and purification of the a-amylase in a laboratory scale
  • Determine the amylolytic and proteolytic activity of enzyme preparations
  • Immobilize proteolytic and amylolytic enzymes by microencapsulation, by crosslinking into the polymer matrices agar and alginate


M. F. Chaplin, C. Bucke (eds.) (2017) Enzyme Technology, Cambridge University Press, Cambridge, New York, Sydney,

M. Kuddus (ed.) (2018) Enzymes in Food Technology, Springer, Singapore (PDF)

K. Buchholz, V. Kasche, U. T. Bornscheuer (2012) Biocatalysts and Enzyme Technology, 2nd ed., John Wiley & Sons, Weinheim

Enzyme Nomenclature. Recommendations on Biochemical & Organic Nomenclature, Symbols & Terminology etc. (

A database for 3-D structures of proteins/enzymes and cofactors important for structure and function ( Enzyme Nomenclature. Recommendations on Biochemical & Organic Nomenclature, Symbols & Terminology etc. (

Recent review and original scientific papers related to research on biotechnological enzyme production and application in biotechnology, food and other industries.

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