1. Adding Value to Molasses Donal Day and C. Chung Audubon Sugar Institute Audubon Sugar Institute Factory Operations Seminar 2007

2. Goal  Upgrade the value of molasses by converting the sugar to a higher value product  Target Market-Quality Liquid Feeds  Others- Human and poultry feeds

3. Background  We have patented a process that produces an oligosaccharide mixture from sugar.  This mixture has shown beneficial effects in animal (poultry) nutrition

4. L. mesenteroides B-742 ( ATCC 13146)  Synthesizes a highly branched dextran from sucrose  High yields of oligosaccharide from sucrose in presence of maltose by acceptor reaction  Fructose conversion to mannitol

5. Oligosaccharide Production-Process Flow Fermentation with L. mesenteroides ATCC 13146 Cell separation Product polishing by chromatography Oligosaccharide product Deionization and Decolorization Mannitol crystallization Input : maltose 50 g/L sucrose 100 g/L, salts Output : oligosaccharides 80.1 g/L (82.3 % theoretical yield) mannitol 37.6 g/L (70.1 % conversion ) 86.4 % mannitol recovered Ca 2+ ion exchange 98.8 % purity

6. Oligosaccharide Distribution by DP % Degree of Polymerization

7. Postulated structures of isomaltooligosaccharides DP, Degree of Polymerization DP3 (panose).  -1,6 o o o o o oo o o o oo o o  -1,4  -1,3 (?) o (glucose).

8. Isomalto(gluco)oligosac charide Process Rapid, simple and economical production method Readily Scalable High production of glucooligosaccharides and mannitol

9. Proposed Mechanism of Action  IMO slows starch utilization (hence growth of undesirable microflora) Growth of desirable microflora favored Intestinal pH drop produces “wash-out” of non-favored species Acts as a  -glucosidase inhibitor, blocking maltose translocation

10. In Vivo Experimentation Toxicity and weight gain in rats LSU Weight gain, Salmonella loading in poultry (broilers) - USDA, Athens, Ga.

11. Rat Toxicity Trials  No toxicity seen  No abnormal physiology on sacrifice  Animals liked the material

12. Body Fat-(12 rats)

13. Blood Triglycerides (12 rats)

16. 1% Nc diet, 2 trials 21 day old poultry (averages) ControlTest Feed Efficiency Weight gain/unit feed 0.580.67 Cecal pH 5.816.63 Salmonella loading cfu/bird x 107 3603.1

17. Estimated Product Value Nutraceutical - unknown  Target market - Poultry Production  Cost attributable to foodborne illness due to poultry about $2 billion/annum  Corn Syrup Replacement  Crude-animal feed supplement  Refinery molasses-healthy baking product Mannitol ($3.32/lb)

18. Costs and Returns (Material) per Kg of Product  Carbon Source (as pure compounds)  Sucrose $0.29  Maltose $0.83  Total $1.12  Products (pure)  Oligosaccharide $9.03 (est)  Mannitol $1.59  Total $9.59

19. Molasses Process  Production of Nc, in situ, in molasses  Process (Blackstrap)  Dilution  Fermentation  Concentration.  Market- QLF

20. Production-Fermentation

21. Process-Pilot Product BlackstrapModified Brix84.171.2 Sucrose % 37.711.5 RS % 15.312.8 IMO % 013.1 Ash % 16.715.9 Proteinndnd

22. What’s Next?  Contacting QLF to set up trials of material in their process.  Investigation of “health benefits” of new material in animal market  Pure Material- pursuing possibility of GRAS status with FDA Research into human food applications

23. Acknowledgement This work was supported in part by the American Sugar Cane League Dr. Roy Martin and Kathleen McCutcheon for their help with the rat studies Dr. Greg Siragusa, USDA, Athens, Ga for his help with the poultry trials