INTER-OFFICE MEMORANDUM Subject! Glucose Isomerase Report : Dote: (Week Ending November 14, 1969) To: Mr. Edward A. Vassallo Pe | = During the production runs of Arthrobacter cells for the pilot ~ plant isomerization test, a lowered yield of enzyme has been observed. Manipulation of the magnesium content of the medium has not solved this problem. ‘The basic problem is the use of the protein hydroly- sates, auch as hydrolyzed animal protein or corn ateep liquor, since these contain many contaminating fons and the composition of these materials’ vary greatly from batch to batch. These probléns are conmon in establishing optimum fermentation conditions and only further experimentation in shake flasks and the 30-gallon fermentors = owl solve these problems. oe i The revised date for the enzymatic conversion experiments in Cedar Rapids is December 8, 1969. The continuous conversion of glucose to fructose was studied using the cell-free enzyme absorbed to DEAE-cellulose in @ short chromatography colunn. The cells were lysed using 1 mg of lysozyme per 25 g of wet weight cells and mechanically disrupting the cells. ‘The enzyme was precipitated with equal volumes of cold acetone at -10C, For this initial experiment with the bound enzyme, 2M glucose — wereused as the initial substrate. Using the same concentration of enzyme that is normally used in a batch conversion and a flow rate of 1 ml/minute, a 42% fructose solution was obtained in about 15 hours. ‘These studies are continuing, since the column approach is a good method for the re-use or continual use of the enzyme. The haze material in the last two syrup conversions (200 gallons) has been renoved by @ powdered carbon treatment. ‘The syrup is presently being concentrated at Product Development. A report on the clean-up procedure and heat stability of these syrups will be issued by the appropriate Product Development personnel. : It was recently observed that the addition of low levels of t cobalt ion to the test system significantly increased the enzymatic activity of the cells. The cobalt and magnesium ions appear to have a synergistic effect on enzymatic activity. The approximate 50% increase in enzymatic activity could reduce or eliminate any additional ion removal costs. The optimum concentration of the ions for maximum enzymatic activity is presently under investigation. ZL90 €€80S CoP: vbi, aoe (See next page for distribution.)Distributs Mr. D. S. Peoples Dr. Murray Senkus Dr. R, B, Farrar Dr. E. D. Nielson Dr. R. E. Brouillard_ Dr. EB. G, Ritchell Mr. W. C. Black Mr. Claude Sawyer : Dr. D. J. Lartigue Dr. J. L. Ayres Dr. Paul Benko Mr. L. E, Hayes Dr.’ Chin K, Lee Miss Margaret E. Long @ 3 3 8 3 s 3