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INTER-OFFICE MEMORANDUM Subject! Glucose Isomerase Report Dote: November 10, 1969 (leek Ending Novenber 7, 1969) CONFIDENTIAL ‘The production of Arthrobacter cells is continuing to obtain enough enzyme to convert approximately 1,000 gallons of high DE syrup as a pilot run in Cedar Rapids. The enzymatic conversion will follow the alkali conversion study since the Product Development personnel will be available to assist in the preparation of the high DE syrup and in monitoring the enzymatic conversion. The tentative date for the initiation of the enzy- matic conversion experiments is November 24, 1969, A concentrated cell-free preparation of isonerase was tested at various temperatures to determine its thermal stability. The enzyme was incubated in the concentrated culture broth for eight hours and little, if any, reduction in enzymatic activity was observed at 40 to 50°C. In the range of 55 to 65°C a slight loss of activity was observed, while at 70°C a significant reduction was observed. However, incubation of the enzyme in the presence of a high DE syrup should improve the temperature stability of the enzyme. ‘The problem of lowered conversion of glucose to fructose using filtered high DE syrup has not been solved. However, it has been observed that low concentrations of iron (107M) inhibit the enzyme. The studies on the various effects of filter aid on enzymatic isomerization are continuing. As noted in last week's report, the studies on re-use of the whole cells are not encouraging. However, it has been observed that the cell free enzyme can be tightly bound to DEAE-cellulose. The enzyme is diffi- cult to remove with water or dextrose’ solutions but can be removed with 0.3M sodium chloride, More economical enzyme carriers, such as carboxy methyl cellulose, are under investigation. Studies are in progress to determine the extent of syrup contamination from the Arthrobacter cells during the conversion of glucose to fructose. It has been observed that the majority of the macromolecular contamination (proteins, nucleic acids, and polysaccharides) occurs during the first two hours of isomerization. ‘he protein constitutes only a small fraction of the contamination. Further characterization of the impurities in the syrup is under investigation to possibly decrease the clean-up costs. 8 ° The effect of a short heat treatment on the crystallization of the enzynatically-isomerized syrups is under investigation. isomerized syrups containing 41%, 43%, 45%, and 47% fructose at 70%, 72%, 742, and 76% solids were given to Dr. A. H. Laurene for a study of their stability at 10°C and 25°C. -2. as the control. cop: vbl Ce: Mr. Dr. Dr. Dr. Dr. Dr. Mr. Mr. Dr. Dre aE. Mr. Dr. Miss Margaret E. Long D. S. Peoples Murray Senkus R, E, Farrar B,D. Nielson R. E, Brouillard E, ¢, Ritchell WC, Black Claude Sawyer D. J. Lartigue J. Le Ayres Paul Benko L. E. Hayes Chin K, Lee Enzymatically— ‘A sample of Tsomerose 100 is used BTTO 96805