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production of fructo-oligosaccharides
from sucrose
Kyung Hoon Jung, Jong Won Yun, Kyung Rae Kang, Jai Yun Lim
and Jae Heung Lee
R & D Centre, Cheil Sugar & Co., Ltd., Kyonggi-Do, South Korea
@1989 Butterworth Publishers Enzyme Microb. Technol., 1989, vol. 11, August 491
Papers
Kinetic studies 100
Unless otherwise specified, enzyme reactions were
carried out for 1 h at 55C and pH 5.5 in test tubes
containing 10 ml of reaction mixture as described 80
Similar reaction patterns were found with other sub- dG 4 180 Vms" S
strates such as 1-kestose and nystose. Sucrose and d---t = 8 342 [Kms + S + (Kms/Kig)G] (2)
Products a
Concentration Molar ratio
Substrate (g 1-1) GF._I (g 1-1) GFn+I (g 1-1) (GF._I/GFn.I)
G 250
F 250
300 10.3 29.2 0.98
GF 400 11.8 34.7 0.96
600 13.4 29.5 1.26
300 1.7 4.3 0.78
GFF 400 2.5 6.6 0.84
600 4.0 9.0 0.86
300 1.0 1.7 1.23
GFFF 400 1.9 2.8 1.10
600 3.5 6.1 0.94
Enzyme reactions were carried out for 20 min instead of 1 h at 55C and pH 5.5
4 x 504 4 G F2 400
300
GF 2o0
2 x 666 2 100
0 10 20 30 40 50 10 20 30 40 50
O F2
Tlmelh)
4 x 828 1 GF 4
Figure 5 Comparison of experimental data with the mathemat-
! ical model at 55C and pH 5.5. (A) 50% sucrose, (B) 65% sucrose.
'. Glucose (O), sucrose (Q), 1-kestose (A), nystose (IlL fructofu-
ranosyl nystose (~)
Figure 4 Network of the proposed reaction mechanism the production of fructo-oligosaccharides by the action
of fructosyltransferase occurs from a consecutive set
of disproportionation reactions. It should be men-
tioned that small amounts of fructose (below 1%) were
The rate of 1-kestose production is complicated: 4
also accumulated slowly as the enzyme reaction pro-
moles of 1-kestose are produced from 8 moles of
gressed. The production of fructose is most likely due
sucrose and 1 mole of 1-kestose is formed from 2
to the action of another enzyme such as invertase as a
moles of nystose. Simultaneously 2 moles of nystose
contaminant in the fructosyltransferase enzyme prepa-
and 2 moles of sucrose are also formed from 4 moles of
ration.
1-kestose.
dK 4 x 504 Vr~s" S
dt 8 X 342 [Kms + S + (Kms/Kig)G] Nomenclature
Vmk " K 504 Vmn " N F fructose concentration (g 1-~)
+ - - (3) G glucose concentration (g l -J)
(Kink+K) 2 x666 (Km.+N) K 1-kestose concentration (g 1-~)
where N indicates nystose, Vm~ indicates Vmax for Kig competitive inhibition constant for glucose
nystose, and Kmn indicates the Michaelis constant for (g 1-1)
nystose. Kink Michaelis constant for 1-kestose (g 1-l)
With respect to the production of nystose, 2 moles Krnn Michaelis constant for nystose (g 1-l)
of nystose are produced from 4 moles of 1-kestose and Krns Michaelis constant for sucrose (g 1-l)
the nystose formed is removed to form 1 mole of N nystose concentration (g 1-l)
fructofuranosyl nystose and 1 mole of 1-kestose. P fructofuranosyl nystose concentration (g l-l)
S sucrose concentration (g 1-1)
dN 2 x 666 Vmk " K Vmn " N t time (h)
- - = - - (4)
dt 4 x 504 (Kmk+ K) (Km. + N) Vmk maximum velocity for 1-kestose (g 1-l h l)
maximum velocity for nystose (g 1-l h -l)
Finally, the rate of fructofuranosyl nystose produc-
Vms maximum velocity for sucrose (g 1-1 h -1)
tion is given by:
dP 828 Vmn " N
d--~- = 66------~
2 x " (Kin, + N) (5) References
where P indicates fructofuranosyl nystose. 1 Hidaka, H., Eida, T., Adachi, T. and Saitoh, Y. Nippon
Nogeikagaku Kaishi 1987, 61, 915-923
The simultaneous integration of the differential 2 Pazur, J. H. J. Biol. Chem. 1952, 199, 217-225
equations describing the proposed mechanism was 3 Gupta, A. K. and Bhatia, I. S. Phytochemistry 1980, 19,
carried out using a digital computer with a fixed step 2557-2563
size of 0.1 h. As illustrated by Figure 5, which shows 4 Gupta, A. K. and Bhatia, I. S. Phytochemistry 1982, 21,
the computer curve and the experimental data, good 1249-1253
5 Smith, J. A., Grove, D., Luenser, S. J. and Park, L. G. US Pat.
agreement was found between the model and the 4 309 505 (1982)
experimental results, although the data points were 6 Jung, K. H., Lira, J. Y., Yoo, S. J., Lee, J. H. and Yoo, M. Y.
scattered to some extent. Therefore, it appears that Biotechnol. Lett. 1987, 9, 703-708