• Microbiology and Biotechnology Letters
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• v.22 no.1
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• pp.106-113
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• 1994

A novel method for production of concentrated purity maltose using swollen extruded corn starch was investigated. Degree of gelatinization of extruded starch suitable for maltose formation was found to be around 70%. The optimal amiunt of enzyme was 400 unit fungal $\alpha $-amylase per g of starch, and the reaction time was 12 hours. At extruded starch concentration of 300 g/l(w/v), maltose concentration and content were reached up to 220 g/l(w/v) and 77%(w/w), respectively. The maltose forming reaction was also successfully proceeded at high starch concentration of 700 g/l(w/v), however, the conversion yield and content were decreased. By the addition of extruded starch by fed-batch wise, the maltose concentration, purity, and conversion yield could be improved up to 465 g/l(w/v), 70%(w/w), and 0.63, respectively. The investigated maltose production process seems to have many potential advantages over the conventional process utilizing liquefied starch, and the feasibility for industrial application needs to be evaluated.

• Archives of Pharmacal Research
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• v.29 no.12
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• pp.1096-1101
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• 2006

Armeniacae Semen not only contains amygdalin, but emulsin also, which is an enzyme that hydrolyzes amygdalin. The extraction yield of amygdalin from Armeniacae Semen was low, due to the presence of emulsin, when extracted with water. When Schizandrae Fructus solution was used as the extractant; however, amygdalin was almost completely extracted, regardless of the cutting size, due to the absence of the influence of emulsin. In addition, when the crude powder or small piece forms were used with Schizandrae Fructus solution, on epimerization of the D-amygdalin into neoamygdalin occurred. D-amygdalin and its conversion product, neoamygdalin, were quantitatively analyzed by reverse-phase, high-performance liquid chromatography (HPLC), with an optimized eluent of 10 mM sodium phosphate buffer (pH 2.3), containing 11.5% acetonitrile. The concentration and detector response were linearly correlated over the range 0.05 to 2 mM. The detection limits for both D-amygdalin and neoamygdalin were approximately $5\;{\mu}M$ for the amount injected.

• Journal of Ginseng Research
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• v.38 no.1
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• pp.47-51
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• 2014

The transformation of ginsenoside Rb1 into a specific minor ginsenoside using Aspergillus niger KCCM 11239, as well as the identification of the transformed products and the pathway via thin layer chromatography and high performance liquid chromatography were evaluated to develop a new biologically active material. The conversion of ginsenoside Rb1 generated Rd, Rg3, Rh2, and compound K although the reaction rates were low due to the low concentration. In enzymatic conversion, all of the ginsenoside Rb1 was converted to ginsenoside Rd and ginsenoside Rg3 after 24 h of incubation. The crude enzyme (b-glucosidase) from A. niger KCCM 11239 hydrolyzed the ${\beta}$-($1{\rightarrow}6$)-glucosidic linkage at the C-20 of ginsenoside Rb1 to generate ginsenoside Rd and ginsenoside Rg3. Our experimental demonstration showing that A. niger KCCM 11239 produces the ginsenoside-hydrolyzing b-glucosidase reflects the feasibility of developing a specific bioconversion process to obtain active minor ginsenosides.

• Journal of Ginseng Research
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• v.38 no.4
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• pp.264-269
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• 2014

Background: In this study, we examined the effects of various enzymes on chemical conversions of ginsenosides in ginseng extract prepared by amylases. Methods: Rapidase, Econase CE, Viscozyme, Ultraflo L, and Cytolase PCL5 were used for secondary enzymatic hydrolysis after amylase treatment of ginseng extract, and ginsenoside contents, skin permeability, and chemical compositions including total sugar, acidic polysaccharide, and polyphenols were determined on the hydrolyzed ginseng extract. Results: Rapidase treatment significantly elevated total ginsenoside contents compared with the control (p < 0.05). In particular, deglycosylated ginsenosides including Rg3, which are known as bioactive compounds, were significantly increased after Rapidase treatment (p < 0.05). The Rapidase-treated group also increased the skin permeability of polyphenols compared with the control, showing the highest level of total sugar content among the enzyme treatment groups. Conclusion: This result showed that Rapidase induced the conversion of ginsenoside glycosides to aglycones. Meanwhile, Cytolase PCL5 and Econase treatments led to a significant increase of uronic acid (acidic polysaccharide) level. Taken together, our data showed that the treatments of enzymes including Rapidase are useful for the conversion and increase of ginsenosides in ginseng extracts or products.

• Journal of Microbiology and Biotechnology
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• v.8 no.5
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• pp.484-489
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• 1998

Galactooligosaccharides (GalOS) were efficiently produced by partially purified $\beta$-galactosidase from the yeast strain Bullera singularis ATCC 24193. Ammonium sulfate precipitation and ultrafiltration methods were used to prepare the enzyme. The enzyme activity decreased at $50^{\circ}C$ and above. A maximum yield of 40% (w/w) GalOS, corresponding to 120 g of GalOS per liter, was obtained from 300 g per liter of lactose solution at $45^{\circ}C$, pH 3.7 when the lactose conversion was 70%. The yield of GalOS did not increase with increasing initial lactose concentration but the total amounts of GalOS did. Volumetric productivity was 4.8 g of GalOS per liter per hour. During this reaction, the by-products, glucose and galactose, were found to inhibit GalOS formation. Reaction products were found to be comprised of disaccharides and trisaccharides according to TLC and HPLC analyses. We propose the structure of the major product, a trisaccharide, to be ο-$\beta$-D-galactopyranosyl-(l-4)-ο-$\beta$-D-galactopyranosyl-(l-4)-$\beta$-D-glucose (4'-galactosyl lactose).

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.4
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• pp.194-200
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• 2002

The cells of Rhodococcus rhodochrous M33, which produce a nitrile hydratase enzyme, were immobilized in acrylamide-based polymer gels. The optimum pH and temperature for the activity of nitrile hydratase in both the free and Immobilized cells were 7.4 and 45$\^{C}$, respectively, yet the optimum temperature for acrylamide production by the immobilized cells was 20$\^{C}$. The nitrile hydratase of the immobilized cells was more stable with acrylamide than that of the free cells. Under optimal conditions, the final acrylamide concentration reached about 400 g/L with a conversion yield of almost 100% after 8 h of reaction when using 150 g/L of immobilized cells corresponding to a 1.91 g-dry cell weight/L. The enzyme activity of the immobilized cells rapidly de-creased with repeated use. However, the quality of the acrylamide produced by the immobilized cells was much better than that produced by the free cells in terms of color, salt content, turbidity, and foam formation. The quality of the aqueous acrylamide solution obtained was found to be of commercial use without further purification.

• BMB Reports
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• v.40 no.2
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• pp.205-211
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• 2007

The stability curve - a plot of the Gibbs free energy of unfolding versus temperature - is calculated for bovine erythrocyte carbonic anhydrase in 150 mM sodium phosphate (pH = 7.0) from a combination of reversible differential scanning calorimetry measurements and isothermal guanidine hydrochloride titrations. The enzyme possesses two stable folded conformers with the conformational transition occurring at ~30$^{\circ}C$. The methodology yields a stability curve for the complete unfolding of the enzyme below this temperature but only the partial unfolding, to the molten globule state, above it. The transition state thermodynamics for the low- to physiological-temperature conformational change are calculated from slow-scan-rate differential scanning calorimetry measurements where it is found that the free energy barrier for the conversion is 90 kJ/mole and the transition state possesses a substantial unfolding quality. The data therefore suggest that the x-ray structure may differ considerably from the physiological structure and that the two conformers are not readily interconverted.

• Membrane Journal
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• v.19 no.1
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• pp.72-82
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• 2009

A mathematical model was written for simulating the removal of phenol from wastewater in enzyme-loaded membrane reactor (EMR). The numerical simulation program was developed so as to predict the degradation of phenol through an EMR. Numerical model proves to be effective in searching for optimal operating conditions and creating an optimal microenvironment for the biocatalyst in order to optimize productivity. In this study, several dimensionless parameters such as Thiele Modulus (${\phi}^2$, dimensionless Michaelis-Menten constant ($\xi$), Peclet number (Pe) were introduced to simplify their effects on system efficiency. In particular, the study of phenol conversion at different feed compositions shows that low phenol concentrations and high Thiele Modulus values lead to higher reactant degradation.

• Journal of Microbiology
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• v.40 no.1
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• pp.33-37
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• 2002

An enzymatic reaction for the production of D-alanine from D-aspartic acid and pyruvate as substrates by a thermostable D-amino acid aminotransferase (D-AAT) was investigated at various conditions In the temperature range of 40-70$\^{C}$ and pH range of 6.0-9.5. The D-AAT was produced with recombinant E. coli BL21, which hosted the chimeric plasmid pTLK2 harboring the D-AAT from the novel thermophilic Bacillus sp. LK-2. The enzyme reaction was shown to follow the Ping Pong Bi Bi mechanism. The K$\_$m/ values for D-aspartic acid and pyruvate were 4.38 mar and 0.72 mM, respectively. It was observed that competitive inhibition by D-alanine, the product of this reaction, was evident with the inhibition constant K$\_$i/ value of 0.1 mM. A unique feature of this reaction scheme is that the decorboxylation of oxaloacetic acid, one of the products, spontaneously produces pyruvate. Therefore, only a catalytic amount of pyruvate is necessary for the enzyme conversion reaction to proceed. A typical time-course kinetic study skewed that D-alanine up to 88 mM could be produced from 100 mM of D-aspartic acid with a molar yield of 1.0.

• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.293-297
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• 2002

The cyclodextrin glucanotransferase (CGTase) gene from Bacillus stearothermophilus NO2 was expressed in Saccharomyces cerevisiae 2805 under the adhl promoter. The CGTase was purified from S. cerevisiae 2805/pVT-CGTS. The purified enzyme exhibited a optima of activity around pH 7.0 and $65^{\circ}C$. Thermal stability of the enzyme was increased fairly as compared with the CGTase of B. stearothermophilus NO2. The conversion yield of cyclodextrin (CD) and the production ratio of $\alpha$-, $\beta$,-, ${\gamma}$-CD from starch were showed similarly aspect to the CGTase of B. stearothermophilus NO2.