1. Lentikats Biotechnology in wastewater treatment

2. Content Our company What is Lentikats Biocatalyst? Advantages of Lentikats Biotechnology Comparison of Lentikats Biotechnology with traditional processes Comparison of Lentikats Biotechnology with traditional processes Key benefits of Lentikats Biotechnology Application of Lentikats Biotechnology Partners References What can we offer? Contacts

3. Lentikats Biotechnology in wastewater treatment The LentiKat’s corporation follows up on a nine year old tradition of the research and development of a new leading edge technology to be integrated into the wastewater treatment industry. We specialise in the efficient elimination of nitrogen pollution. Lentikats Biotechnology has demonstrated a successful elimination of both high and low concentrations of nitrogen from wastewater with the removal efficiency of >98%. We are flexible and competent in preparing custom made solutions.

4. What is a Lentikats Biocatalyst? Unique shape of the matrix and the original principle of the immobilisation enables encapsulation of free enzymes or microorganisms (bacteria, yeasts, fungi) in a stable porous PVA (polyvinyl alcohol) carrier. The carrier’s advantages are: Excellent physical-mechanical characteristics that provide for long-term mechanical stability Low biodegradability Zero toxicity Lentikats Biocatalyst provides for easy separation of enzymes and microorganisms from the surrounding media, thus enabling their multiple reuse. Biotechnologies are changing the world…

5. Carriers Lentikats Biotechnology vs. biomass growth on the carrier surface traditional proven technology new method based on well-proven principle limited working concentration Biocatalyst is highly efficient also at high working concentrations of nitrogen biomass is encapsulated inside the carrier highly active from the beginning, defined composition and known activity uncontrolled development of biomass with variable composition

6. How does Lentikats Biocatalyst function? Due to its maximum surface and at the same time minimum volume of the Lentikats Biocatalyst, the diffusion limitations associated with the transport of the substrate into the carrier and the product out of the carrier are significantly reduced.

7. Nitrification 1.Ammonia nitrogen is oxidised to nitrites by bacteria Nitrosomonas (N. europaea, N. oligocarogenes, …) or Nitrosococcus, Nitrosospira and Nitrosolobus. NH 4 + +1,5 O 2 → NO 2 - + 2 H + + H 2 O 2.Nitrite is oxidised to nitrates by bacteria Nitrobacter (N. agilis, N. winogradskiy) or Nitrospira and Nitrococcus. NO 2 - + 0,5 O 2 → NO 3 - Nitrification – summary: NH 3 + 2 O 2 → NO 3 - + H + + H 2 O

8. Denitrification = one-step biochemical process Denitrifying bacteria (Paracoccus sp., Pseudomonas) utilises nitrate nitrogen as electron acceptor instead of oxygen to create gaseous nitrogen. A red + NO 3 - → N 2 + A ox + energy Denitrification – summary:: A red + NO 3 - → N 2 + A ox + energy Denitrification

9. Lentikats Biotechnology – lay-out

10. Your demands? 1.to meet strict effluent concentration limits for nitrogen 2.to intensify your WWTP without extensive reconstruction 3.to build a new WWTP with reduced operational and investment costs 4.to enhance nitrification and denitrification process of a current treatment system 5.to become more environmentally concerned (reduced energy consumption, lower waste production, lower sludge production…) 6.to simply become better and more efficient in treatment of wastewater

11. Comparison of Lentikats Biotechnology to conventional wastewater treatment technologies 1.Reduction of nitrification and denitrification tank volume. 2.Lower energy consumption, lower organic matter demand, lower oxygen demand in nitrification. 3.Lower waste and sludge production, reduction of sludge management and disposal requirements. 4.High process stability. 5.Ability to respond to even highly fluctuating concentrations. Want to be more efficient? Want to be more environmentally concerned? LentiKat´s a.s. is the right business partner for you.

12. Lentikats Biotechnology benefits 1. reaction volume reduction 1.Higher concentration of nitrification and denitrification organisms. 2.Higher reaction rate - [up to 500-800 mg N (NH 4 +, NO 3 - ) per hour and kg of Lentikats Biocatalyst]. 3.Shorter retention time in nitrification (1-15 hr) and denitrification (0.5 - 2 hr). 4.Reduction of reaction volumes. Activated sludge – 5% nitrifying microorganismsBL – 100% nitrifying microorganisms X

13. Lentikats Biotechnology benefits 2. lower demand for operational material Activated sludgeLentikats Biotechnology Oxygen and organic substrate are utilised only for the biochemical conversion, not for the production of a new biomass. Electricity and heat consumption reduction due to the smaller reaction volumes.

14. microsieve Lentikats Biotechnology benefits 3. lower sludge production AS clarifie r Sludge management Activated sludge system BL Lentikats Biotechnology Bacteria is immobilised inside the Biocatalyst, i.e. there is no need for further production of a new biomass, the actual biomass production is very limited (2 g/m 3 where BOD < 50 mg/l). Sludge production is up to 170 times lower in industrial wastewaters with low content of organic matter.

15. Lentikats Biotechnology benefits 4. high process stability Porous outer shell protects the microorganisms while creating inside „micro-climate“» higher robustness towards negative external effects. High concentration of microorganisms » high working concentration (up to 500-800 mg/l N-NH 4 +, 10 g/l N-NO 3 - ). Long lifetime – 1.5 to 3 years guarantee.

16. Lentikats Biotechnology benefits 5. ability to respond to extensive fluctuations It is possible to set-up and operate the process with a high precision and efficiency of >98% due to the ability to dose the accurate amount of Biocatalyst. Maximum system’s capacity – defined by the amount of Biocatalyst in the reactor with activity corresponding to optimum conditions. Real system’s performance – effectively managed by the control of air/organic substrate dosage. Biotechnologies are changing the world…

17. What do you gain? 1.Costs savings: –no heavy fines incurred for breaking environmental legislation, –operational costs savings (lower organic substrate demand, reduction of costs for sludge management and disposal), –investment costs savings (retrofit of existing WWTPs without extensive reconstruction). 2. Smaller footprint (reduction of reaction volumes). 3. You are more environmentally concerned (lower energy consumption, reduced waste production). 4.Sludge management reduction (up to 170 times lower sludge production*). 5.Reduced operational risks (controlled process, high performance under low temperature, high process stability). * In case of industrial wastewater with low organic matter content..

18. Fields of application Municipal wastewater Intensification of existing wastewater treatment plants without extensive reconstruction to decrease effluent concentration of nitrogen. Industrial wastewater Removal of high concentrations of nitrogen pollution (N-NH 4 +,, N-NO 3 -,, N-NO 2 - ) or of hardly degradable organic compounds. Biogas plants Removal of nitrogen from reject water prior to its discharge into a recipient/WWTP.

19. 1. Intensification of municipal WWTPs by the removal of nitrogen from reject water Option 1

20. 1. Intensification of municipal WWTPs by the removal of nitrogen from reject water System designed to effectively remove nitrogen from reject water, which can make up to 10-20 % of total mass of nitrogen entering the biological unit in conventional municipal WWTPs. Multiple reduction of investment costs in comparison to traditional ways of WWTPs’ intensification – substantially smaller reactors. Applicable for WWTPs with extensive sludge management (anaerobic sludge digestion). Limited applicability depending on the actual content of nitrogen in the reject water.

21. 1. Intensification of municipal WWTPs by the removal of nitrogen from reject water Option 2

22. 1. Intensification of municipal WWTPs by the removal of nitrogen from reject water Thorough analysis of the current system is essential. Evaluation of a denitrification capacity of the primary clarifier and activated sludge system. Cost reduced by the elimination of the denitrification stage – approx. ½ of the Option 1. Denitrification stage using Lentikats Biotechnology can be easily added on at any time. Option 2

23. 2. Intensification of municipal WWTPs by the removal of residual nitrogen from the effluent stream

24. Applicable for any WWTP, which needs to meet its effluent criteria. Mostly reduced to denitrification only in case of conventional municipal WWTPs. Multiple reduction of investment costs in comparison to traditional ways of WWTPs’ intensification – substantially smaller reactors. System works slower at colder temperatures, yet the lifetime of the Biocatalyst is extended by two to three years. It is possible to set up the system for the exact amount of nitrogen to be removed and to further effectively control the system’s efficiency by a simple adjustment of the Biocatalysts’ amount in the system.

25. 3. Removal of high and low concentration of nitrogen from industrial wastewater Simple, compact and highly efficient system. Eliminates the demand for organic substrate in nitrification related to the evolution of a new biomass (in contrast to systems utilising suspended biomass). Substantial reduction of operational costs in comparison to traditional technologies. Possibility to precisely set up the system’s capacity to meet specific requirements. Enhanced resistance towards external factors (fluctuating concentration of nitrogen, pH, presence of toxic compounds, etc.). High system stability, easy operation and control.

26. 4. Nitrogen removal from wastewater with highly fluctuating nitrogen concentration (industry, hotels, petrol stations, etc.) Maximum capacity is defined by the total amount of Lentikats Biocatalyst. Actual performance is effectively managed by the control of air/organic substrate dosage. Simple process automation. High process stability Substantial reduction of operational and investment costs. Biotechnologies change the world.

27. 5. Nitrogen removal from reject water in biogas plants Lentikats Biotechnology allows for working concentration of up to 800 mg N- NH 4 /l, which is almost 4 times higher than what activated sludge systems allow. Removal efficiency of >98% with easy regulation. Applicable for pre-treatment of reject water prior to its discharge into a recipient/WWTP. Pure oxygen application in nitrification alternative. Applicable also as a polishing step after previous treatment using another technology.

28. 6. Special application of Lentikats Biotechnology in wastewater treatment Brines from ion exchange columns regeneration: high concentration of nitrate nitrogen (up to 2.5 g/l N-NO 3 - ) high concentration of inorganic salts (up to 20 g/l NaCl) enables repetitive use of the regeneration solutions (brines) Selective biodegradation: selection, cultivation and immobilisation of suitable microbial cultures to remove hardly degradable organic compounds

29. Lentikats Biotechnology Applications Lentikats Biocatalysts and applied Lentikats Biotechnology also delivers additional value and benefits in the following sectors : distillery – production of bioethanol pharmaceutics – production of robust biocatalysts food industry – whey processing, production of special sugars carbohydrates and syrups Biotechnologies change the world…

30. Our partners in the field of industrial applications Centroprojekt a.s., Štefánikova 167, 760 30 Zlín, Czech Republic www.centroprojekt.cz ASIO s.r.o., Tuřanka 1, 627 00 Brno, Czech Republic www.asio.cz Fermenta Biotech, DIL Complex, Ghodbunder Road, Thane (West) - 400 610, Maharashtra, India www.fermentabiotech.com RUDOS Ružomberok, s.r.o., Štiavnička 190, P. O. Box 112, 034 50 Ružomberok, Slovakia www.rudos.sk geniaLab ®, BioTechnologie-Produkte und Dienstleistungen Gmbh Büchnerstraße 7, 38118 Braunschweig, Germany www.genialab.com

31. Our partners in the field of research and development ICT Prague, Department of Fermentation Chemistry and Bioengineering, Technická 5, 166 28 Prague 6, Czech Republic www.vscht.cz Microbiological Institute, Czech Academy of Sciences, Vídeňská 1083, 142 20 Praha 4, Krč, Czech Republic www.biomed.cas.cz STU Bratislava, Faculty of Food and Biochemical Technology, Institute of Biotechnology and Food Industry, Radlinského 9, 81237 Bratislava, Slovakia www.stuba.sk TUL Liberec, Studentská 2, 461 17 Liberec, Czech Republic www.tul.cz Fermenta Biotech, DIL Complex, Ghodbunder Road, Thane (West) - 400 610, Maharashtra, India www.fermentabiotech.c om University of Jan Evangelista Purkyně, Faculty of Environment, Hoření 13, 400 96 Ústí nad Labem, Czech Republic www.ujep.cz

32. Our references and experience Full-scale application at the WWTP Ostrov u Macochy (1000 PE) for the removal of residual nitrates with a consequent P and COD removal. Full-scale application at the WWTP Litoměřice (40 000 PE) for the nitrogen removal from reject water. Retrofit of the industrial WWTP (4000 EO) for the elimination of fluctuating effluent concentration of nitrates, Baxter Bioscience s.r.o. (pharmaceutical industry) Semi-continuous and continuous pilot-scale nitrification and denitrification of reject water, municipal WWTPs (15 000, 35 000 and 640 000 PE)

33. Why Lentikats Biotechnology? 1.It presents a highly efficient, cost-effective and simple method for removal of nitrogen from wastewater. 2.It is the only economically viable solution for removal of high concentrations of nitrogen. 3.Risk minimisation on investor’s side. We offer financial participation on the pilot-scale test phase of your project. You shall decide about the project execution after the testing period. 4.We guarantee the effluent parameters as designed and approved.

34. What can we offer to you? Have we raised your interest for testing Lentikats Biotechnology in your own project? We are prepared to supply a sufficient amount of our Biocatalyst and to provide full assistance in order to convince you of the obvious advantages our technology has when applied in your wastewater treatment or biogas plant. For all companies introducing new technologies, the first applications and end results are the most important ones. That is why we offer excellent conditions to potential clients in an effort to eliminate any of their risks.

35. Lentikats Biotechnology services Analysis of water influent and effluent parameters. Control of maintenance of specified technological procedures and conditions. Periodical monitoring of Lentikats Biocatalyst’s activity. Replacement of Lentikats Biocatalyst in such periods and volumes to sustain the required Lentikats biotechnology parameters. Disposal of the spent biocatalyst.

36. Contacts Company’s 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