• Food Science and Biotechnology
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• 제17권1호
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• pp.118-122
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• 2008

The use of tartary buckwheat flour as a source of dietary rutin has been limited because of the enzymatic degradation of rutin during the dough-making process, which results in a bitter taste. A variety of pretreatment regimes, including heating, steaming, boiling, and extruding, were evaluated in relation to the inactivation of the rutin-degrading enzyme responsible for rutin loss and color change during dough-making. Steaming (120 see), boiling (90 see) buckwheat grains, or extruding (180 rpm/min at $140^{\circ}C$) the flour resulted in the retention of >85% of the original rutin and eliminated the bitter taste in the hydrated flours. In contrast, dry heating at $140^{\circ}C$ for 9 min or microwaving at 2,450 MHz for 3 min did not reduce the rutin loss, and the bitter taste remained. Unlike in the flour, the rutin degradation in water-soaked grains was insignificant at room temperature. Moreover, the samples treated by steaming, boiling, or extrusion were darker and more reddish in color.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.909-916
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• 2018

Previously, a cytosolic trehalase (TreH) from the hyperthermophilic archaeon Sulfolobus acidocaldarius was reported; however, the gene responsible for the trehalase activity was not identified. Two genes, saci_1816 and saci_1250, that encode the glycoside hydrolase family 15 type glucoamylase-like proteins in S. acidocaldarius were targeted and expressed in Escherichia coli, and their abilities to hydrolyze trehalose were examined. Recombinant Saci_1816 hydrolyzed trehalose exclusively without any help from a cofactor. The mass spectrometric analysis of partially purified native TreH also confirmed that Saci_1816 was involved in proteins exhibiting trehalase activity. Optimal trehalose hydrolysis activity of the recombinant Saci_1816 was observed at pH 4.0 and $60^{\circ}C$. The pH dependence of the recombinant enzyme was similar to that of the native enzyme, but its optimal temperature was $20-25^{\circ}C$ lower, and its thermostability was also slightly reduced. From the biochemical and structural results, Saci_1816 was identified as a trehalase responsible for trehalose degradation in S. acidocaldarius. Identification of the treH gene confirms that the degradation of trehalose in Sulfolobus species occurs via the TreH pathway.

• 환경생물
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• 제34권3호
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• pp.201-207
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• 2016

The hazards associated with the polycyclic aromatic hydrocarbons (PAHs) are known to be recalcitrant by their structure, but white rot fungi are capable of degrading recalcitrant organic compounds. Phlebia brevispora KUC9045 isolated from Korea was investigated its efficiency of degradation of four PAHs, such as phenanthrene, anthracne, fluoranthene, and pyrene. And the species secreted extracellular laccase and MnP (Manganese dependent peroxidase) during degradation. P. brevispora KUC9045 demonstrated effective degradation rates of phenanthrene (66.3%), anthracene (67.4%), fluoranthene (61.6%), and pyrene (63.3%), respectively. For enhancement of degradation rates of PAHs by the species, Remazol Brilliant Blue R (RBBR) was preferentially supplemented to induce ligninolytic enzymes. The biodegradation rates of the three PAHs including phenanthrene, fluoranthene, and pyrene were improved as higher concentration of Remazol Brilliant Blue R was supplemented. However, anthracene was degraded with the highest rate among four PAHs after two weeks of the incubation without RBBR addition. According to the previous study, RBBR can be clearly decolorized by P. brevispora KUC9045. Hence, the present study demonstrates simultaneous degradation of dye and PAHs by the white rot fungus. And it is considered that the ligninolytic enzymes are closely related with the degradation. In addition, it indicated that dye waste water might be used to induce ligninolytic enzymes for effective degradation of PAHs.

• Journal of Microbiology and Biotechnology
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• 제4권2호
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• pp.134-140
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• 1994

An antagonistic bacterium Pseudomonas stutzeri YPL-1 liberated extracellular chitinase and $\beta$-1,3-glucanase which are key enzymes in the decomposition of fungal hyphal walls. The lytic enzymes caused abnormal swelling and retreating at the hyphal tips of plant pathogenic fungus Fusarium solani in a dual culture. Scanning electron microscopy revealed the hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. The production of chitinase and properties of a crude preparation of the enzyme from P. stutzeri YPL-1 were investigated. Peak of the chitinase activity was detected after 4 hr of cultivation. The enzyme had optimum temperature and pH of 50$^{\circ}C$ and pH 5.3, respectively. The enzyme was stable in the pH range of 3.5 to 6.0 up to 50$^{\circ}C$. The enzyme was significantly inhibited by metal compounds such as $HgCl_2$, but was stimulated by $CoCl_2$. P. stutzeri YPL-1 produced high levels of the enzyme after 84 hr of incubation. Among the tested carbon sources, chitin was the most effective for the enzyme production, at the concentration level of 3%. As a source of nitrogen, peptone was the best for the enzyme production, at the concentration level of 4%. The maximum amount of enzyme was produced by cultivating the bacterium at a medium of initial pH 6.8.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.71-71
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• 1995

The succinic semialdehyde dehydrogenase which is one of the key enzyme of GABA shunt in CNS has been purified from bovine brain homogeneously for the first time. The molecular mass of the native enzyme was estimated to be approximately 110,000 on gel filtration, The subunit molecular mass was determined by SDS-PAGE to be 54,000. These results indicate that the enzyme is a dimeric protein made up to identical subunits. Chemical modification studies of the enzyme suggest that the critical lysyl, connected with catalytic activity of the enzyme, The binding of IAF-SSDH(enzyme tagged with fluoreceine) to GABA transaminase which catalyzes the degradation of GABA was monitored by steady emission anisotropy. The changes of fluorescence anisotropy by interactions between two enzymes suggest that the formation of enzyme cluster must be invoved in the regulation of GABA concentration in brain tissues. The inhibitory effects of some antiepileptic and anticonvulsant drugs on the enzyme were also examined.

• Asian-Australasian Journal of Animal Sciences
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• 제24권2호
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• pp.214-221
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• 2011

Effects of the cell wall-degrading enzymes derived from Acremonium cellulolyticus and Trichoderma viride on the silage fermentation and in sacco degradation of tropical grasses i.e. rhodesgrass (Chloris gayana Kunth. cv. Callide) and guineagrass (Panicum maximum Jacq. cv. Natsukaze) were investigated in laboratory-scale experiments. These two grasses were either treated with or without the enzymes before ensiling. Untreated rhodesgrass produced acetate fermentation silage (lactate, $13.0\;g\;kg^{-1}$ DM; acetate, $38.7\;g\;kg^{-1}$ DM) with high final pH value and $NH_3$-N content (5.84 and $215\;g\;kg^{-1}$ DM). Addition of enzymes significantly increased (p<0.01) the lactate production (lactate, 45.6; acetate, $34.0\;g\;kg-^{1}$ DM) and decreased (p<0.01) the pH and $NH_3$-N (4.80 and $154\;g\;kg^{-1}$ DM) in the ensiled forages when compared with the control silages. Untreated guineagrass was successfully preserved with a high lactate proportion (lactate, 45.5; acetate, $24.1\;g\;kg^{-1}$ DM), and the addition of enzymes further enhanced the desirable fermentation (lactate, $57.5\;g\;kg^{-1}$ DM; acetate, $19.4\;g\;kg^{-1}$ DM). The content of NDF was lowered (p<0.05) by enzymes in both silages, but the extent appeared greater in the enzyme-treated rhodesgrass (rhodesgrass, $48\;g\;kg^{-1}$ DM; guineagrass, $21\;g\;kg^{-1}$ DM). Changes in the kinetics of in sacco degradation showed that enzyme treatment increased (p<0.01) the rapidly degradable DM (rhodesgrass, 299 vs. $362\;g\;kg^{-1}$ DM; guineagrass, 324 vs. $343\;g\;kg^{-1}$ DM) but did not influence the potential degradation, lag time and degradation rate of DM and NDF in the two silages.

• Journal of Microbiology and Biotechnology
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• 제23권6호
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• pp.864-871
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• 2013

The recombinant strain P. pastoris GS115-lccC was used to produce laccase with high activity. Factors influencing laccase expression, such as pH, methanol concentration, copper concentration, peptone concentration, shaker rotate speed, and medium volume were investigated. Under the optimal conditions, laccase activity reached 12,344 U/L on day 15. The recombinant enzyme was purified by precipitating and dialyzing to electrophoretic homogeneity, and was estimated to have a molecular mass of about 58 kDa. When guaiacol was the substrate, the laccase showed the highest activity at pH 5.0 and was stable when the pH was 4.5~6.0. The optimal temperature for the laccase to oxidize guaiacol was $60^{\circ}C$, but it was not stable at high temperature. The enzyme could remain stable at $30^{\circ}C$ for 5 days. The recombinant laccase was used to degrade chlorpyrifos in several laccase/mediator systems. Among three synthetic mediators (ABTS, HBT, VA) and three natural mediators (vanillin, 2,6-DMP, and guaiacol), vanillin showed the most enhancement on degradation of chlorpyrifos. Both laccase and vanillin were responsible for the degradation of chlorpyrifos. A higher dosage of vanillin may promote a higher level of degradation of chlorpyrifos, and the 2-step addition of vanillin led to 98% chlorpyrifos degradation. The degradation of chlorpyrifos was faster in the L/V system ($k_{obs}$ = 0.151) than that in the buffer solution ($k_{obs}$ = 0.028).

• Applied Biological Chemistry
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• 제33권2호
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• pp.125-128
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• 1990

저온에서 완두유묘 줄기의 유리 IAA 수준과 그 전구체인 트립토판 수준이 저하하므로, IAA수준의 조절 메커니즘을 규명하기 위하여 IAA분해효소계에 미치는 저온의 영향을 조사하였다. $25^{\circ}C$에서 키운 백색 관두유표를 $5^{\circ}C$의 저온으로 옮기면 IAA oxidase와 peroxidase의 수준은 증가하였으며 catalase의 수준은 저하되었고 과산화수소는 축적되어, 전체적으로 IAA분해계가 저온에서 강화되었다. 저온에 놓였던 유묘를 상온으로 옮기면 IAA 분해계는 다시 대조구 수준으로 회복하였다.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.749-760
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• 2022

α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca²⁺ and Mg²⁺ promoted the activity of T26GAL while Zn²⁺ and Cu²⁺ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

• Asian-Australasian Journal of Animal Sciences
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• 제12권3호
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• pp.358-365
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• 1999

The effects of dry roasting whole lupin seeds (lupinus albus, WLS) at 110, 130 or $150{^{\circ}C}$ for 15, 30 or 45 minutes on the in sacco rumen degradation characteristics, optimal heating conditions of time and temperature and in vitro enzyme digestibility were determined. Ruminant degradation characteristics (RDC) of crude protein (CP) of WLS were determined by in sacco technique in dairy cows. Measure ROC were soluble (S), undegradable (U), potentially degradable (D) fractions, lag time (TO) and rate of degradation (Kd) of insoluble but degradable fraction. Based on measured ROC, percentage bypass CP (%BCP) and bypass CP (BCP in g/kg, DM) were calculated. Degradability of CP was significantly reduced by dry roasting (p<0.001). The interaction of dry roasting temperature and time had significant effects on D (p<0.05), Kd (p<0.01), U (p<0.01), %BCP (p<0.001) and BCP (p<0.001) but not on S (p=0.923>0.05). With increasing time and temperature, S, D, Kd and U varied from 31.8%, 67.4%, 10.3%/h and 0.8% in the raw WLS (RWLS) to 27.1 %, 35.8%, 3.6%/h, 38.4% in $150{^{\circ}C}/45\;min$, respectively. All these effects resulted in increasing %BCP from 25.9 in RWLS to 61.0% in the $150{^{\circ}C}/45\;min$. Therefore BCP increased form 111.2 to 261.2 g/kg DM, respectively. Both %BCP and BCP at $150{^{\circ}C}/45\;min$ increased nearly 2.5 times over the RWLS. The effects of dry roasting on %BCP and BCP seemed to be linear up to the highest value tested. Although ROC had been altered by dry roasting, the In vitro perpsin-cellulase digestibility was generally unchanged. It was concluded that dry roasting was effective in shifting CP degradation from rumen to the lower gastrointestinal tract to potential reduce unnecessary N loss in the rumen. It might be of great value in successfully synchronizing the rhythms of release of nitrogen and energy in the rumen, thus achieving a more efficient fermentation of diets with high proportions of lignocellulosic resources. To determine the optimal dry roasting conditions, the digestibility of each treatment in the cows will be measured in the next trial using mobile bags technique.