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F Cj z:M 2 4 t2 REV .



Subject : Streptomyces Glucose Isomerase Date : December 12, 1969

To : Dr . Murray Senkus

There has been much discussion recently concerning the Japanese

article on glucose isomerase which appeared in Fermentation Advances,
1969 . Much interest has been shown in their report on the re-use of
the enzyme and the heat fixation properties of their enzyme . In order
to clarify a few misconceptions which this article has stimulated, this
short report will discuss the relationships between the Streptomyces
and the RJR glucose isomerase .

Their unit of enzyme activity at 70°C is equivalent to 17 RJR

microunits (}.tU) at 60°C and the maximum activity they reported was
equivalent to about 510 pU/ml (RJR definition) . The yield of Strepto-
_myces cells was approximately 10 grams per liter in 25 to 30 hours,
while the RJR system yields approximately 30 grams per liter in 48 to
55 hours .

The studies on the re-use of the enzyme are"misTeading since they

reported that six units of enzyme were required per gram of glucose
in a batch system . However, 24 units were used per gram of glucose
with heat-fixed cells that were re-used . It is obvious that if one
increases the initial level of enzyme four-fold, one can re-use the
enzyme . In the first four enzyme reactions, syrups of approximately
48, 45, 38, and 43% conversions ~f glucose to fructose were obtained
in 20 hours at 70°C . The enzymatic activity was reduced to approximately
50% remaining after the fifth and sixth reaction ; however, an increased
incubation time (30 to 35 hours) was required to obtain a 40% conversion .
In the seventh reaction, approximately 40 hours were required to produce
a 30% conversion .

In these examples, no mention was made of the color or macro-

molecular contamination in the final syrup With the RJR enzyme, the
time-temperature relationship and enzyme concentration were chosen to
minimize color formation during the isomerization .

CGP :vbl

Cc : Mr . E . A . Vassallo Dr . R . E . Brouillard
Dr . E . D . Nielson Mr . W . C . Black
Mr . C . E . Sawyer