Enzyme production using modern biotechnology

Most commercial enzymes are produced from microorganisms that are enhanced through natural selection, classical strain improvement techniques (e.g., mutagenesis and selection), recombinant DNA technologies and/or gene editing. The Enzyme Technical Association considers modern biotechnology to mean the utilization of genetic engineering, other than selection or classical improvement techniques, to modify organisms for the production of substances. Modern biotechnology may be used in combination with classical techniques

Why do we use biotechnology for enzymes?

The use of classical selection techniques alone poses limitations in that only a small number of strains isolated from nature are able to grow & produce on industrial scale, with few options to improve enzyme expression and with little control over the introduced changes. In addition, the available natural variation in protein sequence poses limits to important characteristics such as pH optimum and temperature stability, especially when seeking specific combinations of such characteristics required under the enzyme application conditions.

Using the modern biotechnology toolbox enables commercialization of a wide diversity of enzymes found in nature that otherwise could not be produced at scale in an economically viable manner, including enzymes from non-culturable microbial sources. The use of targeted methods such as genetic engineering and gene editing also allows for better definition and control of the desired changes than with more traditional, random improvement methods. This results in more consistent enzyme activity/stability in the application and eliminates side activities that may interfere with the application. When synthetic gene sequences are used this allows only the gene for the enzyme to be transferred without transfer of cloning remnants.

Modern biotechnology techniques and why we use them:

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Genetic Engineering/Gene Editing

  • Enabling methods

  • More efficient

  • Biobased and biodegradable

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Protein Engineering Techniques

  • Tailoring to the application

  • New biodiversity

  • Enables new applications

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Classical Selection Techniques

  • Natural & random biodiversity

Use of Safe Strains for Production

By using the tools of modern biotechnology, the enzyme industry has developed safe host organism systems for the production of many enzymes that could not otherwise be produced. These safe host systems have been used since the early 1980’s for the production of different enzymes in contained manufacturing facilities. The host organisms and their enzyme products have been tested to demonstrate that they are safe for their intended use; this includes, but isn’t restricted to, testing of the organism to demonstrate that it is not pathogenic and does not produce toxins, and testing of the product to demonstrate that it is safe for the intended use.

The repeated use of well-established host strains allows for the accumulation of know-how in both strain construction and fermentation operating conditions to maximize predictability in performance and safety of these drop-in production platforms, and the ability to streamline their safety assessment. As a result, enzyme yield and purity are superior for engineered microbes, resulting in a better use of all resources: raw materials, energy, water, land, and less waste, along with a faster route to market, such that society can more readily benefit from the sustainability benefits provided by enzymes.

Benefits of using modern biotechnology

 

  • Safety of production strain is key component to safety evaluation
    • Non-toxigenic, Non-pathogenic
  • Genetic modifications introduced are safe and well characterized
  • Demonstration of History of Safe Use
    • Safety demonstrated by toxicological studies and analysis using well established safety assessment guidelines
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  • Controlled Manufacturing Process
    • Closed manufacturing systems and material transfers to maintain purity and safe enzyme production
    • Fermentation follows aseptic procedures and specific protocols to monitor growth of organism
    • Various parameters (e.g., pH, oxygen, temperature, concentration of nutrients, and cells) are monitored and controlled
    • Quality checks for contamination at critical control points

  • The structure determines an enzyme property such as catalytic activity, specificity, and stability
  • Intentional changes to the amino acid sequence of the enzyme relative to the native sequence allows for enzymes with tailored characteristics
    • Improved thermal stability (e.g., temperature)
    • Improved enzymatic efficiency (e.g., rate of enzyme action)
    • pH optimum for the application conditions
  • Enzyme are selected and sometimes modified to optimize their activity in the intended use application
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  • Follow standard industry practices for purification and formulation
  • Final products meet purity recommendations for enzymes used in intended use applications
  • Optimization of the protein – improve specific activity, purity to meet the intended use of the product application
  • Addition of sequences to improve yield

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To learn more about Biotechnology, click on below:

Enzymes – A Primer on Use and Benefits Today and Tomorrow (PDF)

ETA Position Paper on the Use of Genetic Modification Techniques for the Improvement of Enzyme Producing Organisms and Enzyme Proteins (PDF)

Industrial Biotechnology Journal Publication: “The Generally Recognized As Safe (GRAS) Process for Industrial Microbial Enzymes”