1. Production of Invert Sugar using Lentikats Biotechnology

2. Content Our aim What is Lentikats Biocatalyst
Production of Lentikats Biocatalyst
Benefits of Lentikats Biotechnology
Lentikats Biocatalyst in Food Industry
Our guarantees
Our partners
Contacts 2

3. LentiKat’s a.s. in Food Industry
LentiKat’s a.s. follows up on nine year of research and development of Lentikats Biotechnology.
Our production know-how for commercial use of our Biocatalyst and for its industrial applications is patented internationally.
Our main focus is in intensification of production processes that leads to:
higher process yields
enhanced process stability,
Increased revenue,
Reduction of environmental footprint.
Biotechnologies are changing
the world…

4. 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… 4 4

5. 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 5

6. Why polyvinylalcohol? Biotechnologies are changing the world…
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

7. How is Lentikats Biocatalyst produced?

8. 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

9. Lentikats Biocatalysts Manufacturing Plant

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

11. What are the benefits? Matrix containing high concentration of biomass
Higher yields (in comparison to conventional technologies using free biomass)
Higher reaction rates
Reduction of reaction tanks’ volume
Operational costs savings (electricity, heat)
Possibility of repeated usage
Operational costs savings
Enhanced resistance of modified and immobilised biomass
Higher process stability
Easy separation of biomass from reaction media
Reduced costs for downstream processes (separation)
Lower production of waste and by-products
Environmentally friendly disposal 11

12. ..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… 12 12

13. 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

14. Lentikats and food industry
We currently offer solutions for the following processes:
production of lactose free milk
Lentikats Biocatalyst with β-galaktosidase activity
production of invert sugar
Lentikats Biocatalyst with invertase activity
whey processing
Lentikats Biocatalyst with β-galaktosidase activity for:
production of D-galaktose from whey lactose (pharmaceutical quality)
production of galacto-oligosaccharides from whey lactose
production of special syrups
Lentikats Biocatalyst with invertase activity for production of glucose-fructose syrups
Lentikats Biocatalyst with immobilised glucoamylase for production of glucose from sugar and starch-based materials or glucose syrups from maltotriose or maltose syrup

15. Lentikats and food industry
Following solutions are currently under development and testing:
production of organic acids
Lentikats Biocatalyst with immobilised bacteria Bacillus coagulans for production of lactic acid from lactose, glucose and sucrose
Lentikats Biocatalyst with immobilised filamentous fungi Aspergillus niger for production of gluconic acid from sucrose and glucose
Lentikats Biocatalyst with immobilised filamentous fungi Aspergillus terreus for production of itaconic acid from sucrose and glucose
Biotechnologies are changing the world…

16. Invert Sugar in Food Industry
Sucrose source – sugar beat, sugar cane
Mixture of glucose and fructose (sucrose hydrolysis)
Invert sugar as a product:
composition 33% sucrose, 35% glucose and 32% fructose
» composition 4% sucrose, 51% glucose and 45% fructose
invert sugar is more expensive than syrup
» invert sugar is sweeter and contains also other monosaccharides apart from sucrose
large variety of applications: kids food, pastry, soups and sauces, candy and sweets, fruit drinks, dairy products.

17. Invert sugar in Distilleries
Sucrose source – sugar beat, sugar cane
Mixture of glucose and fructose
molasses – benefits of sucrose hydrolysis
a longer shelf-life
» possibility of a year-round operation for molasses distilleries
a lower water binding capacity – possibility to a achieve higher thickness prior to storage (80%)
» lower demands for storage space and transportation

18. Lentikats Biotechnology and production of invert sugar
Lentikats Biocatalyst with invertase activity
= immobilised saccharase (β-D-fructofuranosidase)
saccharase source – Baker’s yeast (Candida utilis)
invertase activity g sucrose/kg LB/hr
optimum temperature °C
optimum pH 4.5 – 5.0
operational stability 600 – 1000 hodin 18 18

19. Lentikats Biotechnology and production of invert sugar
The active compound is immobilised invertase for sucrose hydrolysis to glucose and fructose.
- Standard activity assay:
batch test
standard sucrose solution with the concentration 100 g/l
temperature 30°C
pH 4.5 (10 mM acetate buffer)
conversion time 50 min
conversion rate 100% 19

20. Case studies

21. 1) Hydrolysis of sucrose solution
Invert sugar production
Input parameters
sucrose concentration 450 g/l
pH = 4.5
temperature = 30°C
batch operation mode
solution volume: m3
Lentikats Biocatalyst
activity 1000 g SUC/kg LB/hr
stability 900 hr
amount 1 ton
Outlet
concentration:
sucrose 10 g/l
glucose 220 g/l
fructose 220 g/l
volume m3
Invert sugar production
amount kg
14 EUR/m3 solution
0.03 EUR/kg invert sugar 21

22. 2) Hydrolysis of sucrose solution
Ethanol production
Input parameters
sucrose concentration 450 g/l
pH = 4.5
temperature = 30°C
batch operation mode
Solution volume: 2222 m3
Lentikats Biocatalyst
activity 1000 g SUC/kg LB/hr
stability 1000 hr
amount 1 ton
Output
concentration:
sucrose 10 g/l
glucose 220 g/l
fructose 220 g/l
volume m3
Ethanol production
volume litres a.a.
14 EUR/m3 solution
0.05 EUR/litre a.a. 22

23. 3) Molasses distillery 4 EUR/m3 molasses 0.05 EUR/litre a.a.
Ethanol production
Input parameters
sucrose concentration 120 g/l
pH = 4.5
temperature = 30°C
continuous operation mode
molasses volume: m3
Lentikats Biocatalyst
activity 1000 g SUC/kg LB/hr
stability 1000 hr
amount 1 ton
Output
concentration:
sucrose 30 g/l
glucose 45 g/l
fructose 45 g/l
volume m3
Ethanol production
volume litres a.a.
4 EUR/m3 molasses
0.05 EUR/litre a.a. 23

24. Summary – application of 1 ton of LB
Sucrose solution – 450 g/l sucrose
Volume of solution processed
[m3]
Ethanol produced
[l a.a.]
Price
[EUR/m3 SUC solution]
[Kč/litr a.a.]
1 ton LB
2222 14
0.05
Molasses – 120 g/l sucrose
Volume of molasses processed
[m3]
Ethanol produced
[l a.a.]
Price
[EUR/m3 molasses]
[Kč/litr a.a.]
1 ton LB
8333 4
0.05

25. 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… 25

26. Investment and Investment Return Guarantees
We guarantee the following:
output parameters
operational stability of Lentikats Biocatalyst and its activity and productivity

27. Our partners in the field of industrial applications
Fermenta Biotech, DIL Complex, Ghodbunder Road, Thane (West) , Maharashtra, India
Protean, Pod Lesem 9, Dobra Voda u Ceskych Budejovic, Czech Republic
Pharmix s.r.o., Na Sadkach 2798, Kromeriz, Czech Republic 27 27

28. Our partners in the field of research and development
Fermenta Biotech, DIL Complex, Ghodbunder Road, Thane (West) , Maharashtra, India
Institute of Microbiology, Academy of Science of the Czech Republic Vídeňská 1083, Prague 4, Krč, Czech Republic
STU Bratislava, Fakulta chemické a potravinářské technologie, Ústav biotechnologie a potravinářství, Radlinského 9, Bratislava, Slovakia
VŠCHT Praha, Ústav kvasné chemie a bioinženýrství, Technická 5, Praguea 6
University of Alberta, Edmonton Alberta, T6G 2J9 Canada
Masaryk University, National Centre for Biomolecular Research, Kamenice 5, Brno-Bohunice 28 28

29. Thank you for attention
Laboratory:
INBIT LentiKat‘s a.s.
Kamenice 34
Brno
Czech Republic
Tel.:
Tel.:
Company Headquarters:
Office Building ARGO
LentiKat‘s a.s.
Evropska 846/176a
Prague 6
Czech Republic
Tel.:
Fax:
Production plant:
Pod Vinicí 83
Stráž pod Ralskem
Tel.:
Fax: 29 29