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P 4 .~'ii.t 2 ;?.L RE Y . 5 .


~ '~N ~ I r- ~ ~~ I A L

Subject : Glucose Isomerase Report Date : January 7, 1970

. (Period Covering Dec . 22, 1969 to
Jan . 2, 1970)

To : Mr . Edward A . Vassallo

r- The glucose isomerase activity in shake flask cultures has been

raised to about 700 pU per ml by a modificati .on in inoculation procedures .
These results will be verified this week in the 30 gallon fermentors using
the three different inoculation procedures .

, The procedures for the reuse of the enzyme have been studied using
the column method . An enzyme concentration equivalent to a 700 ml batch
of syrup was bound to DEAE-cellulose . The bound enzyme was inserted in a
jacketed glass column and the column was heated by passing water at 60°C
t hrough the column jacket . In these preliminary experiments, 2M glucose
slightly buff ered at pH 8 .0 and containing a magnesium salt was used as
the substrate, and approximately 700 ml of isomerized syrup were collected
f per day .
After 16 days, the enzyme was still active and approximately 45%
conversions of glucose t.o fr ;uctose were obtained . *The columns will be
run until exhaustion so that an approximate measure of enzyme stability
can be obtained . Although these are preliminary data, the potential of
this procedure is very significant . In comparing these data with a batch
enzymatic isomerization, the partially purified enzyme is being reused 16
times with potential for further reuse . Although the color units and ion
load were not determined, the % :olor of the isomerized syrup was comparable
to the initial syrup . The studies will be repeated using filtered high
DE syrups ; and variables such as pH, temperature and flow rates will be
examined .

Charles G . Pheil

CGP :gof

Cc : Mr . D . S . Peoples Mr . Claude Sawyer

Dr . Murray Senkus Dr . D . J . Lartigue
Dr . R. E. Farrar Dr . J . L . Ayres
Dr . E. D. Nielson Dr . Paul Benko
Dr . R. E. Brouillard Mr . L . E . Hayes
Dr . E. C. Ritchell Dr . Chin K . Lee
Mr . W . C . Black Miss Margaret E . Long