Application of protein engineering
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This document discusses various applications of protein engineering in different industries and fields. It describes how protein engineering is used in the food industry to modify enzymes like proteases, amylases, and lipases to improve their properties. It also discusses applications in environmental remediation, medicine like cancer treatment, biopolymer production, nanobiotechnology, and redox proteins. The document provides an overview of the wide range of uses of protein engineering across diverse domains.
Application of protein engineering
- 1. APPLICATION OF PROTEIN ENGINEERING AKSHAY PARMAR Post Graduate Student | Department Of Biochemistry School Of Science, Gujarat University.
- 2. WHAT IS PROTEIN ENGINEERING ? • Protein engineering can be defined as the modification of protein structure with recombinant DNA technology or chemical treatment to get a desirable function for better use in medicine, industry and agriculture. • Protein engineering is the process of developing useful or valuable proteins. It is a young discipline, with much research taking place into the understanding of protein folding and recognition for protein design principles. It is also a product and services market, with an estimated value of $168 billion by 2017.
- 3. FOOD INDUSTRY • Important application area of protein engineering regarding food industry is the wheat gluten proteins. Their heterologous expression and protein engineering has been studied using a variety of expression systems, such as E.coli, yeasts or cultured insect cells. • Wildtype and mutant wheat gluten proteins were produced to compare them to each other for protein structure-function studies because of their availability, rapid and easy use, aswell as high expression levels. • Food industry makes use of a variety of food-processing enzymes, such as amylases and lipases, the properties of which are improved using recombinant DNA technology and protein engineering. The deletion of native genes encoding extracellular proteases, for example, increased enzyme production yields of microbial hosts.
- 4. • Some large groups of enzymes like Proteases, Amylases and Lipases are important for both food and detergent industries, as they have a broad range of industrial applications. • Proteases are used in several applications of food industry regarding low allergenic infant formulas, milk clotting and flavors. They are also important for detergent industry for removing protein stains. • The improvement of proteases for industry to have, for example, high activity at alkaline pH and low temperatures, or improved stability at high temperatures is a challenge for protein engineering. • Microbial protease production is industrially suitable because of low costs, high production yields , and easy genetic manipulation. • Amylases are also important for both food and detergent industries. In food industry, they are use for liquefaction and saccharification of starch, as well as in adjustment of flour and bread softness and volume in baking.
- 5. • The detergent industry makes use of amylases in removal of starch stains Recently, the production of “functional foods” is becoming increasingly important for food industry. Particularly, the production of industrial products and functional foods from cheap and renewable raw agricultural materials is desirable. • Conversion of starch to bioethanol or to functional ingredients like fructose, wine, glucose and trehalose, requires microbial fermentation in the presence of biocatalysts such as amylases to liquefy and saccharify starch. To improve the industrially important properties of amylases, such as high activity, high thermo- and pH-stability, high productivity, etc.; recombinant enzyme technology, protein engineering and enzyme immobilization have been used. • Another major group of enzymes utilized by food and detergent industries is constituted by Lipases.
- 6. • Lipases are used in many applications of food industry such as for the stability and conditioning of dough (as an in situ emulsifier), and in cheese flavour applications. • Lipases are also crucial for the detergent industry, as they are used in removal of lipid stains As lipases are commonly used in food industrial applications, having toxicologically safe lipases is an important requirement of food industry.
- 7. ENVIROMENTALAPPLICATIONS • Environmental applications of enzyme and protein engineering are also another important field. • Genetic methods and strategies for designing microorganisms to eliminate environmental pollutants and included gene expression regulation to provide high catalytic activity under environmental stress conditions, such as the presence of a toxic compound, rational changes introduced in regulatory proteins that control catabolic activities, creation of new metabolic routes and combinations. • Many organic pollutants such as phenols, azo dyes,organophosphorus pesticides and polycyclic aromatic hydrocarbons can be detoxified using enzymatic oxidation.
- 8. • Petroleum biorefining is also an important environmental application area, where new biocatalysts are required. • Protein engineering, isolation and study of new extremophilic microorganisms, genetic engineering developments are all promising advances to develop new biocatalysts for petroleum refining. Petroleum biorefining applications such as fuel biodesulfurization, denitrogenation of fuels, heavy metal removal, depolymerisation of Asphaltenes. • Many protein engineering strategies were identified such as improvement of hydrogen peroxide stability, increasing the redox potential to broaden the substrate range, heterologous expression and industrial production development.
- 9. MEDICALAPPLICATION • The use of protein engineering for cancer treatment studies is a major area of interest. Pretargeted radioimmunotherapy has been discussed as a potential cancer treatment. By pretargeting, radiation toxicity is minimized by separating the rapidly cleared radionuclide and the long-circulating antibody. • Advances in protein engineering and recombinant DNA technology were expected to increase the use of pretargeted radioimmunotherapy. • The use of novel antibodies as anticancer agents is also an important field of application, where the ability of antibodies to select antigens specifically and with high affinity is exploited, and protein engineering methods are used to modify antibodies to target cancer cells for clinical applications. • Recently, the term “modular protein engineering” has been introduced for emerging cancer therapies.
- 10. • Treatment strategies based on targeted nanoconjugates to be specifically directed against target cells are becoming increasingly important. Additionally, multifunctional and smart drug vehicles can be produced at the nanoscale, by protein engineering. These strategies could be combined to identify and select targets for protein-based drug delivery. • Pharmacokinetic properties of antibodies have been improved by protein engineering and antibody variants of different size and antigen binding sites have been produced for the ultimate use as imaging probes specific to target tissues. • Molecular imaging tools based on antibodies will find more applications in the future regarding diagnosis and treatment of cancer and other complex diseases.
- 11. BIOPOLYMER PRODUCTION • Protein engineering applications for biopolymer production are also promising. Particularly,peptides are becoming increasingly important as biomaterials because of their specific physical, chemical and biological properties. • Protein engineering and macromolecular self assembly are utilized to produce peptide-based biomaterials, such as elastin-like polypeptides, silk- like polymers. • The ability of protein engineering to create and improve protein domains can be utilized for producing new biomaterials for medical and engineering applications.
- 12. NANOBIOTECHNOLOGY • Nanobiotechnology applications of protein engineering are becoming increasingly important. The synthesis and assembly of nanotechnological systems into functional structures and devices has been difficult and limiting their potential applications for a long time. • Biological macromolecules, such as proteins, carbohydrates and lipids are used in the synthesis of biological tissues in aqueous environments and mild physiological conditions, where this biosynthetic process is under genetic regulation. • Particularly proteins are crucial elements of biological systems, based on their roles in transport, regulation of tissue formation, physical performance and biological functions. Thus, they are suitable components for controlled synthesis and assembly of nanotechnological systems.
- 13. • Combinatorial biology methods commonly applied in protein engineering studies, such as phage display and bacterial cell surface display technologies, are also used to select polypeptide sequences which selectively bind to inorganic compound surfaces, for ultimate applications of nanobiotechnology. • Another interesting nanotechnology application is the use of amyloid fibrils as structural templates for nanowire construction. This application is based on the fact that some proteins form well-ordered fibrillar aggregates that are called amyloid fibrils. • The self association of well-ordered growth fibrils through noncovalent bonds under controlled conditions was suggested to have a high potential to be used for nanobiotechnology. The use of amyloid fibrils as structural templates for nanowire construction.
- 14. REDOX PROTEINS AND ENZYMES • Such proteins and enzymes can be modified to be used in nanodevices for biosensing, as well as for nanobiotechnology applications. • The electrochemistry of redox proteins particularly draws attention for applications in biofuel cells, chemical synthesis and biosensors. • Protein engineering of redox-active enzymes pointed out two emerging areas of protein engineering of redox-active enzymes:novel nucleic acid-based catalyst construction, and intra-molecular electron transfer network remodelling. • Cytochrome P450 monooxygenases and protein engineering of cytochrome P450 biocatalysts for medical, biotechnological and bioremediation applications.
- 15. OTHER NEW APPLICATION • “Insertional protein engineering” applications are also becoming important, particularly for biosensor studies. The applications of insertional protein engineering for analytical molecular sensing. • “Zinc finger protein engineering” is another approach that has been used in gene regulation applications. The zinc finger design and principle is used to design DNA binding proteins to control gene expression. • “Virus engineering” is another emerging field, where the virus particles are modified by protein engineering. Viruses have many promising applications in medicine, biotechnology and nanotechnology. They could be used as new vaccines, gene therapy and targeted drug delivery vectors, molecular imaging agents and as building blocks for electronic nanodevices or nanomaterials construction.
- 16. • “Protein cysteine modifications” are also important protein engineering applications. As cysteine modifications in proteins cause diversities in protein functions, cysteine thiol chemistry has been applied for in vitro glycoprotein synthesis. • This method could be potentially used for development of new protein- based drugs, improving their half-life, reducing their toxicity and preventing multidrug resistance development.
- 17. In Many Biological Structures Proteins Are Simply Components Of Larger Molecular Machines. -Michael Behe (American Biochemistry)
- 18. Thank YOU