• Journal of Aquaculture
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• v.18 no.4
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• pp.245-251
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• 2005

This study was investigated the condition of their maximum activity to assay the enzymes of rotifer, Brachionus rotundiformis의 $\alpha$-amylase, total alkaline Protease, trypsin and TG-lipase activities of rotifer were higher and more sensitive in phosphate-NaOH buffer than Tris-HCl buffer. $\alpha$-amylase, trypsin and TG-lipase activities were appeared the maximum at pH 8.0, and total alkaline protease activity showed the maximum activity at pH 7.0. $\alpha$-amylase activity showed the highest activity at $40^{\circ}C$, and total alkaline protease and trypsin activities were assayed the highest at $55{\~}60^{\circ}C$. However, TG-lipase activity was appeared the highest at $25{\~}30^{\circ}C$. The optimum substrate concentration of enzyme activity of a-amylase, total alkaline protease, rypsin and TG-lipase were $3.5\%$ starch, $\0.6%$ azo-casein, $87.5{\mu}M$ BApNA and 81.2 mM olive oil, respectively. The optimum reaction time of enzyme activity of $\alpha$-amylase, total alkaline protease, trypsin and TG-lipase were increased up to 40, 60, 30 and 25 min., respectively. The data obtained in this study could be used for the digestive enzyme research of rotifer, B. rotundiformis.

• Korean Journal of Microbiology
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• v.31 no.2
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• pp.175-178
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• 1993

Alkaline .alpha.-amylase expressed in the transformant, Baciollus subtills MB809, containing alkaline amylase gene cloned from alkalophilic Bacillus sp. AL-8, was purified through for step separation processes. The purified alkaline .alpha.-amylase had molecular weight of app[roximately 59, 000 daltons on SDS-PAGE and Sephaex G-100 gel filtration. Amino acid sequence of terminal portion of the enzyme was analyzed with pure amylase eluted form the SDS-PAGE gel. N-terminal amino acid sequence of .alpha.-amylase was determined by the Edman degradation method and resulted in $NH_{2}$-ser-thr-ala-pro-ser-(ile)-lys-ala-gly-thr-(ile)-leu. For C-terminal amino acid sequencing, purified .alpha.-amylase was digested with carboxypuptidase A and B, and reverse-phase HPLC gradient elution system resulted in -thr-trp-pro-lys-COOH.

• Journal of environmental and Sanitary engineering
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• v.23 no.1
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• pp.25-31
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• 2008

A bacterial strain, Bacillus megaterium L-49 has been isolated and identified that produces alkaline ${\alpha}$-amylase. The cell is ellipsoidal, about $1.0-1.2{\times}3.0-3.6{\mu}m$ in diameter, Gram-positive, motile, and central partial central. Growth occurs in media containing 7% of NaCl. This strain could utilize D-glucose, lactose, xylose, sucrose, mannose, and maltose, and but it does not utilize D-fructose, and glycogen. Among the various concentrations of saturated ammonium sulfate, the retractation ratio in range of 70 to 100% was about 93%. However, in the case of acetone, it was about 98.7%. EDTA has activating effect and Ca2+ has no effect on alkaline ${\alpha}$-amylase activity. The alkaline ${\alpha}$-amylase has low thermal stability. The optimal temperature for reaction is $50^{\circ}C$. The alkaline ${\alpha}$-amylase activity maintained stabilizing at pH 6-11 and the optimal pH for reaction was 9-10.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.403-408
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• 1985

Enhancement of surfactant on release of secretory enzyme, such as $\alpha$-amylase, acid phosphatase and alkaline phosphatase, was investigated during submerged culture of Rhizopus oryzae. Morphological changes of colony was occured; small pelletal form in 0.18mM of sodium dodecyl sulfate, pulpy form in 0.48mM of sodium deoxycholate, and filamentous form in absence of surfactant. It. Supplement of sodium dodecyl sulfate induced 9 times increasing activity of $\alpha$-amylase and that of acid phosphatase 25 times in cultural fluids. Alkaline phosphatase was increased 11 times in cultural fluid and also stimulated in cytoplasm with supplement of sodium deoxycholate.

• Microbiology and Biotechnology Letters
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• v.6 no.1
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• pp.9-16
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• 1978

Effects of the addition of Ginseng extract and it's incubation time on enzyme activity (${\alpha}$-amylase, ${\beta}$-amylase, protease) of Aspergillus app. was investigated. 1. ${\alpha}$-Amylase activity of Asp. oryzae was higher for 48 hours than control, however, the same after 48 hours. ${\beta}$-Amylase activity was stimulated at the concentration of 0.1 to 1.0％, and decreased at the concentration of 3.0 to 5.0％. The acid protease activity was higher than control for 72 hours and in the medium of 120 hours was decreased significantly. The alkaline protease activity was lower than control. However, alkaline protease activity was higher than acid protease activity. 2. ${\alpha}$-Amylase of Asp. niger was increased in proportion to increasing of the extract concentration. ${\beta}$-Amylase was increased at the concentration from 0.5 to 3.0％ and it's activity was depressed in proportion to increasing of the extract concentration for 48 hours and on the contrary it was improved in proportion to increasing of the extract concentration for 72 hours except for 5.0％ concentration with marked decreasing. Alkaline protease activity was increased at lower concentration (0.1∼0.5％) for 24 to 48 hours and it was depressed at higher concentration (1.0 to 5.0), however after 48 hours incubated, it's activity was decreased in preparation to increasing of the extract concentration.

• Journal of Microbiology and Biotechnology
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• v.2 no.3
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• pp.166-173
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• 1992

The expression of Bacillus subtilis $\alpha$-amylase from the phoA-amyE fusion gene in recombinant E. coli was investigated under various environmental conditions. The overexpression of cloned $\alpha$-amylase caused retardations in cell growth and synthesis of alkaline phosphatase (AP) from the chromosomal phoA gene. The change of culture temperature from $37^\circ{C}$ to $30^\circ{C}$ increased the specific activities of both $\alpha$-amylase and $\beta$-lactamase by six and two times, respectively, whereas the AP activity remained unchanged. The experiments with chlorampenicol (a translation inhibitor) suggested the enhancement of $\alpha$-amylase activity at $30^\circ{C}$, and this was partly due to the stability of $\alpha$-amylase itself. The further decrease of the temperature to $25^\circ{C}$ slowed down both the cell growth and cloned-gene expression rate. The $\alpha$-amylase activity showed a maximum at pH of 7.4 while alkaline phosphatase was most effectively produced at pH of 8.3.

• The Korean Journal of Mycology
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• v.25 no.2 s.81
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• pp.85-90
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• 1997

The Isozyme activities of Lentinula edodes were studied as a preliminary study for genetic analysis after protoplast fusion. The presence of peroxidase, esterase, superoxide dismutase, acid phosphatase, alkaline phosphatase, alcohol dehydrogenase and ${\alpha}-amylase$ was examined. An intracellular buffer-soluble protein from the mycelia was used for enzyme analysis on nondenaturing polyacrylamide gels. The auxotrophs of Lentinula edodes were positive for peroxidase, esterase, superoxide dismutase and acid phosphatase. However, alkaline phosphatase, alcohol dehydrogenase and ${\alpha}-amylase$ were not detected. The esterase and peroxidase were not affected by the various culture age. Isozyme identification may be a useful tool after protoplast fusion.

• KSBB Journal
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• v.13 no.5
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• pp.621-625
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• 1998

The raw starch hydrolysis by amylase prepared from alkaline-tolerant Bacillus sp. were investigated. Degree of hydrolysis(%) of 5%(w/v) raw rice, corn and potato starch by this enzyme were about 40, 25 and 20%, respectively. The hydrolysis action on raw starch by change of blue value was similar to the action pattern of exo ${\beta}$-amylase. The hydrolysis products of rice starch were mainly glucose and maltose. Oligosaccarides were also detected. From the above results, this enzyme was considered as exo type ${\alpha}$-amylase. This enzyme activity on the raw starch and the gelatinized starch were 28.40 and 86.60 IU/mg protein, respectively, and the ratio of raw starch-digesting activity to gelatinized starch-digesting activity (raw starch digestivity) was about 32%. The Km values for the raw and the gelatinized starch were 4.22 and 3.0mg/mL, respectively, and the VmaX values were 0.20 and 0.31mg/mL/min, respectively.

• Preventive Nutrition and Food Science
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• v.6 no.2
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• pp.101-106
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• 2001

Alpha-amylase and glucoamylase activities were higher in koji with 40% water than that with 30 and 50% water, and A. oryzae exhibited very high alpha-amylase and glucoamylase activities compared to A. sojae and A. niger. Acidic, neutral and alkaline protease activities also showed higher activities in koji prepared with flour, Korean wheat powder and soybean powder with 40% water based on the weight of the sample. Alpha-amylase, acidic, neutral and alkaline protease activities of all the koji samples according to incubation periods increased until 3~4 days of incubation and maintained nearly the same level or slightly decreased after 5 days of incubation. The protease activities of A. oryzae and A. sojae showed nearly the same trend regardless of differences in substrate conditions and koji materials, but those of A. niger showed a lower activity than those of A. oryzae and A. sojae. These results suggest that the preparation of koji is possible with Korean wheat powder and soybean powder and A. sojae can be utilized as a new strain for fermented foods using soybean as the main materials to increase functional properties and produce products having a new taste and flavor.

• Journal of Aquaculture
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• v.21 no.3
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• pp.190-195
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• 2008

We investigated the changes in growth, digestive enzymes activities, nucleic acids contents and RNA/DNA ratio of flounder Paralichthys olivaceus larvae (C for Paracyclopina nana, A for Artemia, and M for Mix of C and A) for 14 to 28 DAH. Body length of flounder larvae showed the best in the C trial at 28 DAH. The change of nucleic acids contents showed faster in C and M trials than A trial. And RNA/DNA ratio showed the significantly faster changes in C trial than A trial. High metamorphosis rates were also observed in C and M trial. $\alpha$-amylase activities increased gradually up to 28 DAH in all trials. Total alkaline protease (TAP) activities of A trial showed the highest value to 9 mU/larvae at 26 DAH. But others trials showed lower to $5{\sim}6$ mU/larva than A trial. TAP:$\alpha$-amylase activity ratio did not significantly changed to $0.025{\sim}0.053$ in A trial during the experiments. But, C and M trials tended to gradually decrease from $0.078{\sim}0.083$ (initial) to $0.013{\sim}0.018$ (final). Therefore, it shown the ratio gradually decreased of TAP:$\alpha$-amylase activity, stability of TAP activity, and rapid change of nucleic acids in trials grown positively. Thus, because P. nana could continuously supply the optimal nutrients for flounder larvae, we suggested the supplement of the copepod to an efficient feed of the flounder larvae.