Lentikats B iotechnology in O rganic A cid P roduction

1. Lentikats Biotechnology in Organic Acid Production

2. Content What is the Lentikats biocatalyst? How is the Lentikats biocatalyst produced? Benefits of the Lentikats biotechnology Lentikats biocatalyst and its usage in organic acid production Partners Guarantees Contacts

3. What is the Lentikats Biocatalyst? Unique shape of the matrix and original immobilization procedure enables to encapsulate free enzymes or microorganisms (bacteria, yeasts, fungi) inside a solid porous carrier from polyvinylalkohol (PVA). The advantages of PVA cells 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. Biotechnologies are changing the world…

4. 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.

5. Why polyvinylalcohol? 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…

6. How is the Lentikats Biocatalyst produced?

7. What can be immobilized? Free or immobilized enzymes The size of Lentikats biocatalyst pores varies depending on the size and diameter of the biocatalyst due to: type of PVA additives (size of macromolecules) concentration of additives biomass concentration • Free cells • Dead cells • Living non-growing cells • Living growing cells

8. Laboratory Units for theEncapsulation Lab scale units with a low production capacity of Lentikats biocatalysts (for research purposes) are also available

9. Lentikats Biocatalyst – production line

10. What are the benefits of LB? Biotechnologies that are environmentally friendly …

11. …and also… Lower biochemical reaction time In comparison with conventional technologies much higher yield (i.e. 5 – 15% higher) Reuse of the biocatalyst (sieve separation) Reduction of the risk of contamination during fermentation processes High stability of production processes

12. Lentikats Organic Acid Production Solutions Lentikats biotechnology is used in the following processes: • lactic acid production from glucose, sucrose and lactose (immobilized bacteria Bacillus coagulans) • gluconic acid production from sucrose and glucose (immobilized filamentous fungi Aspergillus niger) • itaconic acid production from sucrose and glucose (immobilized filamentous fungi Aspergillus terreus).

13. Lentikats Biotechnology LENTIKATS BIOTECHNOLOGY FOR LACTIC ACID PRODUCTION Producer: bacteria Bacillus coagulans. Substrates: glucose, sucrose, lactose. Production: • batch, fed batch and continuous lactic fermentation is stable for all C-substrates. • saccharose fermentation solutions with60 g/l of saccharose concentrate(The total fermentation process was decresed from 48 hours in„the first conversion“ to 12 hours in „the tenth conversion“with a lactic acid yield of 91 to 93%.) • glucose fermentation solution with 60 g/l glucose concentrate (After 14 hours fermentation broths consist of 55.2 g /l of lactic acid and 1.2 g /l of glucose in the tenth conversion.) • fermentationproductivity is increased from 13.5 to 17.0 (g LA.dm-3.h-1).

14. Lentikats Biotechnology LENTIKATS BIOTECHNOLOGY FOR ORGANIC ACIDS PRODUCTION Producer: 1) filamentous fungi Aspergillus terreus for itaconic acid production 2) filamentous fungi Aspergillus niger for gluconic acid production Substrates: sucrose, glucose Production: • extensive production of itaconic acid is not stable yet • first results indicate that if the initial concentration of sucrose is 50 g/l then the concentration of itaconic acid is 18.5 g/l in the batch mode process

15. Optically clear L-lactic acid (99%) manufacturing Fermentation processes regulation What does Lentikats Biocatalyst bring? Biomass regeneration costs decrease Robustness High yield of lactic acid (about 20%) High specific activity Operational costs decrease

16. Significant improvement of operating economic results Notable cost reduction Low risk of contamination Delivering on Lentikats Biotechnology Key Benefits Decreased fermentation volumes Increased operational costs Decreased sales revenue High yields About 91-93 % High Productivity Short fermentation time(10 hours) Reusable Lentikats Biocatalysts High concentration of Lentikats Biocatalysts in fermentors

17. Our partners in the field of industrial applications

18. Lentikats Biotechnology – guarantee of your investments and the investment return • You can test our biocatalyst easily by yourself with your substrate inyour own plant or plant of your choiceto see the results. • We offer youour assistance and full support. • We offer a long-term cooperation to the engineering companies specialising in the design of distilleries and are interested in the implementation of technologies requiringminimal investment at high profitability. Biotechnologies are changing the world…

19. We guarantee for the following: • delivery of production facilities within 24 months • delivery ofoutput parameters • operational stability of Lentikats biocatalyst and theiractivity and productivity • purchase price stability within 36 months. Lentikats Biotechnology – Investment and Investment Return Guarantees

20. Our partners in the field of research and development

21. Lentikats Biotechnology Applications Lentikats Biocatalysts and applied LentikatsBiotechnology also deliversadditional value and benefits in the following sectors: waste and industrial water treatment, distillery industry – bioethanol production, pharmaceutics – production of robust biocatalysts, food industry – whey processing, production of special sugars carbohydrates and syrups Biotechnologies are changing the world…

22. Contacts Company Headquarters: LentiKat‘s a.s. Office Building ARGO Evropska 846/176a 160 00 Prague 6 Czech Republic Tel.: + 420 255 710 680 Fax: + 420 255 710 699 E-mail: info@lentikats.eu Production plant: 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 Laboratory: INBIT LentiKat‘s a.s. Kamenice 34 625 00 Brno Czech Republic Tel.: + 420 511 205 288 Tel.: + 420 511 205 287