• BMB Reports
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• v.37 no.4
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• pp.422-428
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• 2004

Raw-starch-digesting $\alpha$-amylase (Amyl III) was purified to an electrophoretically pure state from the extract of a koji culture of Aspergillus awamori KT-11 using wheat bran in the medium. The purified Amyl III digested not only soluble starch but also raw corn starch. The major products from the raw starch using Amyl III were maltotriose and maltose, although a small amount of glucose was produced. Amyl III acted on all raw starch granules that it has been tested on. However, it was considered that the action mode of the Amyl III on starch granules was different from that of glucoamylase judging from the observation of granules under a scanning electron microscope before and after enzyme reaction, and also from the reaction products. Glucoamylase (GA I) was also isolated and it was purified to an electrophoretically pure state from the extract. It was found that the electron micrographic features of the granules after treatment with the enzymes were quite different. A synergistic effect of Amyl III and GA I was observed for the digestion of raw starch granules.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2001.06a
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• pp.40-45
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• 2001

Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.

• Microbiology and Biotechnology Letters
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• v.21 no.5
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• pp.461-467
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• 1993

Transglycosylation reaction of cyclodextrin glucanotransferase (CGTase) was analyzed in the attrition coupled heterogeneous reaction system using raw starch as a donor` and mono-, di-saccharide, and glycoside as acceptors. For transglycosylation reaction of stevioside, the transglycosylation rate was similar and the transglycosylation yield was increased compare with conventional process using liquefied starch as the donor. Also the accumulation of maltooligosaccharides in reaction mixture was minimized.

• Natural Product Sciences
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• v.15 no.3
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• pp.134-138
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• 2009

In an attempt to establish anti-ulcerogenic activity of potato tubers, inhibitory activity against ethanol- and indomethacin-induced gastric ulcer models in rats was evaluated for the first time. From several varieties of potato tubers including Solanum tuberosum L. cv. Superior (white skin and fresh potato) and two new varieties of (Bora valley and Gogu valley), raw potato juice was prepared and the starch was obtained from each juice by filtration and drying. Upon oral administration to rats, raw potato juice showed more or less inhibitory activity. The starch showed higher and dose-dependent inhibitory activity, suggesting that the active ingredient in raw potato juice may be the starch. Particularly, the starch obtained from the tubers of new potato variety, "Bora valley," with purple color, showed the highest inhibitory activity (62.4% and 37.1% inhibition of ulcer index at 500 mg/kg), while omeprazole (proton pump inhibitor) used as a reference drug showed 74.4% and 75.7% inhibition at 20 mg/kg against ethanol- and indomethacin-induced ulcer formation, respectively. The present study provides a first evidence of anti-ulcerogenicity of raw potato juice and the starch. Especially, the starch from "Bora valley" strongly inhibited ulcer formation in rats. Considering that these are food components, they may be safely used for anti-ulcerogenic nutraceuticals.

• Applied Biological Chemistry
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• v.32 no.4
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• pp.374-377
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• 1989

To optimize the enzymatic saccharification of raw-starch, reaction conditions by shaking with glass beads were adapted together with ${\alpha}-amylase$ from Streptomyces sp. 4M-2 and amyloglucosidase from commercial source. When raw-starch was degraded by the ${\alpha}-amylase$ in shaking flask with beads (raw-starch : bead in diam. of 3mm=1 : 5 by weight) at the shaker speed of 300rpm, the saccharification rate of corn and potato starch were increased up to 88% and 69% after 30 hrs of reaction, respectively. Application of the amyloglucosidase in combination with the ${\alpha}-amylase$ enhanced the rate of saccharifcation: 88% of saccharification was obtained in 6 hrs for raw-corn starch under the same reaction conditions as above.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.383-390
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• 2010

A newly isolated active producer of raw-starch-digesting amyloytic enzymes, Rhizopus microsporus var. chinensis CICIM-CU F0088, was screened and identified by morphological characteristics and molecular phylogenetic analyses. This fungus was isolated from the soil of Chinese glue pudding mill, and produced high levels of amylolytic activity under solid-state fermentation with supplementation of starch and wheat bran. Results of thin-layer chromatography showed there are two kinds of amyloytic enzymes formed by this strain, including one $\alpha$-amylase and two glucoamylases. It was found in the electron microscope experiments that the two glucoamylases can digest raw corn starch and have an optimal temperature of $70^{\circ}C$. These results signified that amyloytic enzymes secreted by strain Rhizopus microsporus var. chinensis CICIM-CU F0088 were types of thermostable amyloytic enzymes and able to digest raw corn starch.

• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.252-258
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• 1994

Transglycosylation of stevioside in the attrition coupled heterogeneous reaction system using raw starch as a glycosyl donor has significant advantages over conventional reaction systems using liquefied starch as a donor. The transglycosylation of stevioside under the presence of organic solvent showed that transglycosylation reaction occurs via two steps ; initially from raw starch to cyclodextrin(CD), and then followed by transglycosylation of produced CD. Comparison of the transglycosylation efficiency of c$\alpha $-, $\beta $, $\gamma $-CDs indicated that $\alpha $-, $\beta $-CD are mainly utilized as a glycosyl donor for following reaction. The reaction mechanism of transglycosylation between stevioside and CD proceeded according to random sequential bireactant mechanism. The equilibrium constant of transglycosylation reaction of cyclodextrin glucanotransferase wase also evaluated. The structure of transglycosylated stevioside was confirmed by TLC, and it was found that glycosyl group(G$_{1}, $ ~ G$_{4}$-glycosidic bond.

• Culinary science and hospitality research
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• v.20 no.4
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• pp.310-321
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• 2014

This study investigated the effects of starches(potato, sweet potato, and corn starch) on the quality characteristics of raw and cooked noodles. The moisture content, color, mixogram, texture profile and sensory evaluation were performed. The mixogram showed the addition of starches shortened the kneading time. Mixing tolerance was higher in the noodles with starch than the control group, which indicates that the addition of starch increases dough durability. Moisture content of raw noodles was the highest in the samples with potato starch. L-value of raw noodles was the highest in the samples added with corn starch, whereas that of cooked noodles was the highest in the samples added with potato starch. The hardness of raw noodles was the highest in the control sample, and the chewiness of raw noodles was the highest in the samples added with sweet potato starch. The hardness of cooked noodles was the highest in the samples added with corn starch and the lowest in the control group. The tension distance of raw noodles was the longest in the control group and the tension force of raw noodles was the highest in the noodle with sweet potato starch. The acceptance test indicated that the noodles prepared with the addition of sweet potato starch scored the highest in flavor, taste, texture and overall acceptability.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.718-726
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• 2010

A selected fungal strain, for production of the raw-starchdigesting enzyme by solid-state fermentation, was improved by two repeated sequential exposures to ${\gamma}$-irradiation of $Co^{60}$, ultraviolet, and four repeated treatments with Nmethyl-N'-nitrosoguanidine. The mutant strain Aspergillus sp. XN15 was chosen after a rigorous screening process, with its production of the raw-starch-digesting enzyme being twice that of usual wild varieties cultured under preoptimized conditions and in an unsupplemented medium. After 17 successive subculturings, the enzyme production of the mutant was stable. Optimal conditions for the production of the enzyme by solid-state fermentation, using wheat bran as the substrate, were accomplished for the mutant Aspergillus sp. XN15. With the optimal fermentation conditions, and a solid medium supplemented with nitrogen sources of 1% urea and 1% $NH_4NO_3$, 2.5 mM $CoSO_4$, 0.05% (v/w) Tween 80, and 1% glucose, the mutant Aspergillus sp. XN15 produced the raw-starch-digesting enzyme in quantities 19.4 times greater than a typical wild variety. Finally, XN15, through simultaneous saccharification and fermentation of a raw rice corn starch slurry, produced a high level of ethanol with $Y_{p/s}$ of 0.47 g/g.

• Applied Biological Chemistry
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• v.28 no.1
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• pp.1-7
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• 1985

Exo-maltotetraohydrolase produced by Pseudomonas stutzeri IAM 12097 was characterized with respect to substrate specificity, the reaction products and hydolysis rate on various carbohydrates. Maltopentaose, maltoheptaose, soluble starch, amylose, amylopectin, oyster glycogen and gelatinized starch of corn, potato, glutinous rice, green banana and arrow root were hydolyzed by this enzyme, but ${\alpha},{\beta},{\gamma}-cyclodextin$, sucrose, raffinose, lactose, pullulan, maltose, maltotriose and maltotetraose were not hydrolyzed. Among oligosaccharides, maltohexaose was favorably hydrolyzed by this enzyme and the main reaction product of oligosaccharides and polysaccharides was maltotetraose. Addition of pullulanase to this enzyme increased the hydolysis rate on gelatinized starches. tut it did not on raw starches. Among various starches, corn starch was favorably hydrolyzed by this enzyme, whereas it acted on potato starch, arrow root starch and high amylose corn starch weakly.