‘Foil Me nev. 5/07 Subject: Glucose Isomerase Report Deter Novenber 4, 1969 (Week Ending October 31, 1969) Tot Mr, Edward A, Vassallo Since it has been decided to make large-scale enzymatic isomerizations (500 to 1,000 gallons) in Cedar Rapids, a trip was made to the R. J. Reynolds Avoca Farm to determine if any of the chemical reactors could be converted to fermentation vessels, There ate four reaction vessels (2 of 500 gallons, i 1 6£ 1,000 gallons, and 1 of 2,500 gallons) which could probably be converted at minimum expense to'fermentors. A few successful runs in these vessels could prepare enough cells to isomerize many thousand gallons of syrup for better experimental clean-up studies and market test evaluations. The idle chemical plant also contains storage and hold tanks, pumps, a 20-square-foot Bird-Young vacuum filter and a Sharples/Fletcher basket-type centrifugal filter. : Five fermentations were successful in this period.” ‘the four piodictica ~~ runs yielded cells containing glucose isomerase activities of 690 to 780. units per tl Im the experimental run, the yeast extract was replaced with corn steep liquor and cells containing about 600 » units/ml were obtained. Excess foaming was observed in some of these fermentations and this foaming could not be attributed to the antifoam used or the mediun componi “Since foaming must be controlled for an efficient fermentation, some experi- mentation must be performed during the production runs and variables such as aeration-agitation rates and back-pressure in the ferméntors are under investigation. Another 100-gallon batch of enzyme-isomerized syrup was prepared for clean-up studies and for blending with last week's batch for feeding trials. ‘The initial substrate was an unfiltered high DE syrup and, after conversion, a syrup containing approximately 43% fructose was obtained. These syrups a¥é p¥ederitly being filtered and cleaned up by the Product Development ] “—~personnel. A portion of this batch was centrifuged to remove the celle prior __ to filtration while the remainder was only filtered through Standard Super Cei to remove the cells. In both cases a slight haze was stili observed in the syrup and studies of the procedures to remove this haze are under investigation. A laboratory convereion was performed to investigate the possible © use of the cells. The initial one-liter batches of syrup, using fresh cells, contained approximately 35% fructose after 12 hours and 43% after 24 hours. | The cells were then removed from the syrup by centrifugation and resuspended in one liter of fresh syrup (equivalent to, 1’ part cella to 1 part ayrup: and into 500 ml syrup (equivalent to, 2:1 ratio cells to syrup). After 12 9290 ceeeshours of incubation the syrup with the ratio of 1:1 contained 16,5% fructose while the 1:2 ratio syrup contained 26% fructose. These values were correct for the zero time controle. At 12 hours the isomerization curves were : essentially flat which suggests very little, if any, increase in fructose content upon further incubation, Further studies on the effect of various filter aids on enzymatic activity and re-use of the cells are under tavesttgatto D. 8. Peoples Murray Senkus R, E, Farrar E, D. Nielson R. E, Brouillard E. C, Ritchell W. C, Black Claude Sawyer D. J. Lartigue J. Ly Ayres Paul Benko L. E. Hayes Chin K. Lee Miss Margaret E, Long 4490 eeses