• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.323-330
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• 1998

Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0～9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.

• The Korean Journal of Mycology
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• v.31 no.2
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• pp.59-66
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• 2003

Laccase produced by Pycnoporus cinnabarinus SCH-3 isolated from Korea was partially purified using ultrafiltration, anion exchange chromatography and affinity chromatography, The laccase was produced as the predominant extracellular phenoloxidase during primary metabolism. Neither lignin peroxidase nor manganese-dependent peroxidase were detected in the culture fluid. In order to examine the effect of inducers in laccase production, 2,5-xylidine was added in the culture of Pycnoporus cinnabarinus SCH-3. Addition of 2,5-xylidine enhanced 25-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 9%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to be $44.4{\mu}M\;and\;56.0{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $60{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 10 min at $80{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the P. cinnabarinus SCH-3 laccase showed 94% of homology to the N-terminal sequences of laccases from P. cinnabarinus PB and P. coccineus.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.2
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• pp.170-174
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• 1999

Proessing conditions for fermented and dried sauces with the underutilized fishes were investigated. Hair tail, gizzard shad, and kangdale were hydrolyzed at $60^{\circ}C$ for 6 hours using $4\%$ Alcalase, and their hydrolysates were separted by molecularporous membrane. The hydrolytic ratios of hair tail, gizzard shad, and kangdale were estimated to be $84.2\%$, $83.6\%$ and $85.1\%$, respectively. Amino nitrogen recoveries were determind to be $73.1\~73.9\%$ by a membrane with molecular weight cutoff 100 dalton and $91.7\~92.5\%$ by a membrane with 500 dalton. Ultrafiltration was very efficient means for removing bitter taste. With the additions of $2\%$ glucose, $4\%$ lactose and $4\%$ skim milk, product yields of hair tail, gizzard shad, and kangdale were determind to be $16.4\%,\;17.2\%$ and $17.0\%$, respectively. Water adsorption rates of hair tail and kangdale showed $5.0\~9.2\%$ and $5.5\~9.6\%$, respectively, under Aw 0.52$\~$0.94. Contents of total nitrogen in the fermented and dried sauces prepared with hair tail, gizzard shad and kangdale were $3.9\%,\;4.1\%$ and $3.7\%$, respectively, and those of amino nitrogen were $3.2\%,\;3.4\%$ and $3.1\%$, respectively. In the fermented and dried sauces prapared with hair tail, gizzard shad and kangdale, the hygroscopities at Aw 0.88 were $6.9\%,\;7.5\%$ and $6.8\%$, respectively, and solubilities under dissolved in water for 30 minutes were $84.6\%,\;83.6\%$ and $93.8\%$, respectively.