Escolar Documentos
Profissional Documentos
Cultura Documentos
568-575
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Youssef Ben Ammar*, Takayoshi Matsubara, Kazuo Ito, Masaru Iizuka,
Tipaporn Limpaseni, Piamsook Pongsawasdi and Noshi Minamiura
Laboratory of Enzyme Chemistry, Graduate School of Science, Osaka City University,
3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558, Japan
Introduction
569
570
Protein
(mg)
Total activity
(U)
Specific activity
(U/mg)
Yield
(%)
Purification fold
Crude enzyme*
Alcohol precipitation
1st DEAE-Cellulose
1st P-100
2nd DEAE-Cellulose
2rd P-100
3th P-100
38000
1200
920
364
240
40
27
700
700
780
560
570
500
412
0.018
0.58
0.85
1.54
2.37
12.5
15.2
100
100
111
80
81
71.4
58.8
1
32.2
47.2
85.5
131.7
694.5
844.5
*: Two liters of culture medium were used to generate the crude enzyme.
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G1
G2
G3
Others
(G4+G5)
0
0.25
0.5
1
2
4
24
72
0
0
0
0
0
0
8.2
11
0
28.2
47.8
54.6
56
58
75
75
0
31.2
31.8
29.8
29
27.5
4.7
2.8
0
40.5
20.4
15.6
15
14.5
12.1
11.2
for all of these maltooligosaccharides, except for the longtime incubations (more than 24 h) in which glucose appeared
in small amounts when compared to the expected amount to
be released. It was also remarkable for all of these digestions
that glucose appeared while the amount of maltotriose was
decreasing. This result led us to suggest that glucose may
mainly come from the hydrolysis of maltotriose. However, the
Km values of this enzyme towards maltohexose,
maltopentaose, malototetrose, and maltotriose were 26.7,
30.1, 20.5, and 357 mM, respectively. This low affinity to
maltotriose is one of the main differences between this
enzyme and the one from P. expansum, which was reported to
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DQGPDOWRWHWUDRVH&G (A) Final products from 1% starch after a 24-h hydrolysis with 13 units of purified enzyme at 30oC, pH 6.0. M1:
(a mixture of G1, G2, G3, and G4); 1: spot of the enzyme reaction mixture.G (B) Different products from 2% fructosyl-maltotrioside
after hydrolysis with 1 unit of purified enzyme at 30oC, pH 6.0 for different incubation times (0 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, and 24
h). M1: (a mixture of G1, G2, G3, and G4), M2: (a mixture of F1, F2, F3, F4, F5, and F6), M3: sucrose.G (C) Different products from
2% maltotetraose after hydrolysis with 1 unit of purified enzyme at 30oC, pH 6.0 for different incubation times (0 h, 0.25 h, 0.5 h, 1 h,
2 h, 4 h, and 24 h).
573
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Table 3. Softness of the bread during the time after the addition
of the maltose-producing -amylase
Softness of the bread (g/cm2)
Control*
G2 -amylase**
After 1 day
After 3 days
After 7 days
181
156
293
252
373
322
*Without enzyme
**With maltose-producing -amylase
References
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Fogarty, W. M. (1983) Microbial amylases; in Microbial enzyme
and biotechnology, Fogarty, W. M. (ed.), pp. 1-92, Applied
Science Publishers, London.
Goldberg, J. D. and Edwards, C. (1990) Purification and
characterization of an extracellular amylase from a thermophilic
streptomycete. Appl. Bacteriol. 69, 712-717.
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