1. Lentikats Biotechnology in Pharmaceutics

2. Content  Our focus  Protein engineering  What is Lentikats Biocatalyst  Production of Lentikats Biocatalyst  Benefits of Lentikats Biotechnology  Lentikats Biocatalyst in Pharmaceutics  Our guarantees  Our know-how, references and experience  Our partners  Contacts

3. LentiKat’s a.s. in Pharmaceutics Our main focus in the pharmaceutics is on production of robust biocatalysts by application of the following scheme: 1)molecular cloning and production of recombinant proteins, 2)proteins’ activity and stability enhancement by application of protein engineering methods, 3)immobilisation using Lentikats Biotechnology Our approach leads to: higher process yields, enhanced process stability, improved process economics.

4. Protein engineering We offer molecular design and production of any enzyme in a recombinant host system. Using state-of-the-art protein engineering technology we gain: Higher enzymatic activity Improved robustness of the enzyme Our partners in the field: National Centre For Biomolecular Research, Masaryk University Protean s.r.o. Biotechnologies are changing the world…

5. Lentikats Biotechnology Unique shape of the matrix and original immobilisation procedure enables to encapsulate free enzymes or microorganisms (bacteria, yeasts, fungi) inside a hydrogel porous carrier from polyvinyl alcohol (PVA). The advantages of PVA matrix are: excellent physical and mechanical characteristics that result in a long-term mechanical stability, hardly biodegradable, no toxic effects. Lentikats Biocatalysts enable easy separation of enzymes or microorganisms from the bulk media and therefore can be used repeatedly. With biotechnologies to a better future…

6. Microencapsulation Lentikats Biotechnology spherical shape – limited diffusion to the “core” traditional proven technologynew method based on well-proven principle lentil-like shape – no diffusion limitations „micro“ size„mili“ size – easier separation vs.

7. How does the Lentikats Biocatalyst work? A maximum surface in combination with the minimum volume of Lentikats Biocatalyst results in a substantial reduction of diffusion limitations associated with the transfer of substrate into the matrix and the product out of the matrix.

8. Why polyvinylalkohol? It is an inexpensive material Encapsulation takes place under optimal, cell-sparing conditions It allows a production of hydrogel with excellent mechanical properties It does not have any side effects on fermentation It offers high capacity for the cell loading The process is stable and easy to control It is a non-toxic material Biotechnologies are changing the world…

9. How is the Lentikats Biocatalyst produced?

10. Lentikats Biocatalysts Biocatalyst Types for Use in Lentikats Encapsulation: Free Cells dead cells viable, non growing cells viable and growing cells. Free or Immobilised Enzymes. Lentikats Biotechnology allows to immobilise any type of enzyme The size of Lentikats Biocatalyst pores varies depending on the size and diameter of the Biocatalyst due to: PVA additive type (size of macromolecule) additive concentration biomass concentration.

11. Lentikats Biocatalysts Manufacturing Plant

12. Laboratory Units for Encapsulation For low-capacity production of encapsulates (especially for research purposes), a laboratory device is available.

13. What are the benefits? Higher reaction rates Reduction of reaction tanks’ volume Operational costs savings (electricity, heat) Lower production of waste and by-products Environmentally friendly disposal Matrix containing high concentration of biomass Higher yields (in comparison to conventional technologies using free biomass) Possibility of repeated usage Operational costs savings Enhanced resistance of modified and immobilised enzyme Higher protein resistance and activity durability Immobilisation prevents enzyme aggregates disruption Reduced costs for downstream processes (separation)

14. ..and also… Shortening of the reaction times Much higher yields in comparison to conventional technologies (up to 5 – 15 % higher) Repetitive use of the Biocatalyst, easy separation from media (sieve separator) Higher productivity of the Biocatalyst Lower risk of contamination during the production process High stability and easy control of production process Biotechnologies are changing the world…

15. Technological set-up  - inlet of reaction media  - filling of Lentikats Biocatalyst  - medium with Lentikats Biocatalyst  - stirrer  - sieve separator  - outlet of separated media  - outlet of Lentikats Biocatalyst     

16. Lentikats Solutions Lentikats Biotechnology is used in the following pharmaceutical processes: 6-aminopenicillanic acid production (6-APK) (immobilized enzyme with activity of penicillin G-acylase) 7-aminodeacetoxycephalosporanic acid production (7-ADCK) (immobilized enzyme with penicillin G-acylase activity) Following products are available for chemical industry: production of D-galactose from lactose (immobilised enzyme β -galactosidase, p.a.) production of galacto-oligosaccharides from lactose (immobilised enzyme β -galactosidase)

17. Lentikats Solutions Products that are currently under development and being tested Amoxicillin synthesis (synthesis of amoxicillin from p- hydroxyphenylglycine methyl ester and 6-aminopenicillanic acid by immobilized penicillin G acylase) production of lactic acid from glucose, sucrose and lactose (immobilised bacteria Lactobacillus coagulans) production of 1,3 – propanediol (immobilised bacteria Clostridium butyricum) production of dihydroxyaceton (immobilised bacteria Gluconobacter oxydans)

18. Lentikats Solutions for Antibiotics Production LENTIKATS BIOTECHNOLOGY FOR PRODUCTION OF ANTIBIOTICS Enzyme: immobilised penicillin G-acylase for production of antibiotics. Specific Activity: specific activity is determined by use of standard solution and conditions in batch test: - pen G: 8% w/v - temperature: 28°C - pH: 8.0 (5 mM phosphate buffer) - conversion time: 80-120 min - conversion rate: 98-99% Operation Stability: 600-800 repeated batch conversions. sourceactivity E.coli70 U / g LB wet weight 380 U / g LB dry weight

19. Investment and Investment Return Guarantees You can easily verify our great results by testing our Biocatalyst in your or other facilities with the material of your choice. Sample of Lentikats Biocatalyst free of charge. Our company provides assistance and full technical support. Our company offers a long-term cooperation to companies, whose main concerns are reduced investment costs and maximal process profitability. Biotechnologies change the world…

20. Investment and Investment Return Guarantees We guarantee the following: 1.delivery of production facilities and devices within 24 months 2.output parameters 3.operational stability of Lentikats Biocatalyst and its activity and productivity

21. Our know-how, references and experience in pharmaceutics Production and supply of Lentikats Biocatalyst with Penicillin G-acylase activity for production of antibiotics

22. Our partners in the field of industrial applications Fermenta Biotech, DIL Complex, Ghodbunder Road, Thane (West) - 400 610, Maharashtra, India www.fermentabiotech.com Protean, Pod Lesem 9, 373 16 Dobra Voda u Ceskych Budejovic, Czech Republic www.protean.cz Pharmix s.r.o., Na Sadkach 2798, 767 01 Kromeriz, Czech Republic www.pharmix.cz

23. Our partners in the field of research and development Fermenta Biotech, DIL Complex, Ghodbunder Road, Thane (West) - 400 610, Maharashtra, India www.fermentabiotech.com Institute of Microbiology, Academy of Science of the Czech Republic Vídeňská 1083, 142 20 Prague 4, Krč, Czech Republic www.biomed.cas.cz STU Bratislava, Fakulta chemické a potravinářské technologie, Ústav biotechnologie a potravinářství, Radlinského 9, 81237 Bratislava, Slovakia www.stuba.sk VŠCHT Praha, Ústav kvasné chemie a bioinženýrství, Technická 5, 166 28 Praguea 6 www.vscht.cz University of Alberta, Edmonton Alberta, T6G 2J9 Canada www.ualberta.ca Masaryk University, National Centre for Biomolecular Research, Kamenice 5, 625 00 Brno-Bohunice www.ncbr.chemi.muni.cz

24. Contacts Company Headquarters: LentiKat´s a.s. Evropská 423/178 160 00 Praha 6 Czech republic Tel.:+ 420 224 362 461 Fax:+ 420 224 362 463 E-mail:info@lentikats.eu Production plant and laboratories LentiKat´s a.s. Pod Vinicí 83 471 27 Stráž pod Ralskem Czech republic Tel.:+ 420 487 851 393 Fax:+ 420 487 851 399 E-mail: info@lentikats.eu