• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1811-1817
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• 2007

Extracellular enzymes from Lentinus edodes M290 on normal woods (Quercus mongolica) and waste logs from oak mushroom production were comparatively investigated. Endoglucanase, cellobiohydrolase, ${\beta}$-glucosidase, and xylanase activities were higher on waste mushroom logs than on normal woods after 1. edodes M290 inoculation. Xylanase activity was especially different, with a three times higher activity on waste mushroom logs. When the waste mushroom logs were used as a carbon source, a new 35 kDa protein appeared. After the purification, the optimal pH and temperature for xylanase activity were determined to be 4.0 and $50^{\circ}C$, respectively. More than 50% of the optimal xylanase activity was retained when the temperature was increased from 20 to $60^{\circ}C$, after a 240 min reaction. At $40^{\circ}C$, the xylanase maintained 93% of the optimal activity, after a 240 min reaction. The purified xylanase showed a very high homology to the xylanase family 10 from Aspergillus terreus by LC/MS-MS analysis. The highest Xcorr (1.737) was obtained from the peptide KWI SQGIPIDGIG SQTHLGSGGS WTVK originated from Aspergillus terreus, indicating that the 35 kDa protein was xylanase. This protein showed low homology to a previously reported L. edodes xylanase sequence.

• Mycobiology
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• 제43권1호
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• pp.81-86
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• 2015

To promote the selection of promising monokaryotic strains of button mushroom (Agaricus bisporus) during breeding, 61 progeny strains derived from basidiospores of two different lines of dikaryotic parental strains, ASI1038 and ASI1346, were analyzed by nucleotide sequencing of the intergenic spacer I (IGS I) region in their rDNA and by extracellular enzyme assays. Nineteen different sizes of IGS I, which ranged from 1,301 to 1,348 bp, were present among twenty ASI1346-derived progeny strains, while 15 different sizes of IGS I, which ranged from 700 to 1,347 bp, were present among twenty ASI1038-derived progeny strains. Phylogenetic analysis of the IGS sequences revealed that different clades were present in both the ASI10388- and ASI1346-derived progeny strains. Plating assays of seven kinds of extracellular enzymes (${\beta}$-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease) also revealed apparent variation in the ability to produce extracellular enzymes among the 40 tested progeny strains from both parental A. bisporus strains. Overall, this study demonstrates that characterization of IGS I regions and extracellular enzymes is useful for the assessment of the substrate-degrading ability and heterogenicity of A. bisporus monokaryotic strains.

• Preventive Nutrition and Food Science
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• 제14권3호
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• pp.252-259
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• 2009

We previously isolated Bacillus strains with high fibrinolytic activities (FAs) from cheonggukjang prepared by traditional ways. To test their potential as starters for cheonggukjang, soybean was fermented for 72 hr at $37^{\circ}C$ with each isolate and a control lab strain: B. subtilis CH3-25 (BS3-25), B. amyloliquefaciens CH51 (BA51), B. amyloliquefaciens CH86-1 (BA86-1), and B. subtilis 168 (BS168, control, lab strain). Viable cell numbers of all cheonggukjang samples rapidly increased and reached about $10^9$ CFU/g after 6 hr. During 72 hr, the initial pH of 6.3 rapidly increased to 8.1$\sim$8.2 for cheonggukjang fermented with BS3-25 or BA86-1, and 7.3 for those with BA51 or BS168. FAs and protease activities (acid, neutral, and alkaline) rapidly increased in cheonggukjang fermented with BS3-25, BA51, or BA86-1 during the first 12 hr. On the other hand, those of cheonggukjang fermented with BS168 slightly increased during the first 36 hr. There were significant changes in acid and neutral protease activities in cheonggukjang fermented with BA51 or BA86-1 during the 24 hr. Rapid increases of $\beta$-glucosidase activity corresponded well with rapid increases of $\alpha$-amylase and $\alpha$-galactosidase activities in addition to increases in antioxidant activities and the TPCs (total phenolic contents). The highest increase in the TPCs was observed in cheonggukjang fermented with BA86-1 while the least was that fermented with BS168.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2012년도 정기총회 및 춘계학술발표회
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• pp.8-8
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• 2012

Ginseng have been traditionally used for strengthening immunity, providing nutrition and recovering health from fatigue. Recently, pharmaceutical activities of ginseng roots have been proven by many researches, and ginseng has become a world-famous medicinal plant. Ginseng saponin, ginsenoside, is one of the most important secondary metabolite in ginseng which has various pharmacological activities. Many studies have aimed to convert major ginsenosides to the more active minor ginsenoside Rg3 for consumer demand ginseng product. Microbial strain GS514 strain was isolated from soil around ginseng roots for enzymatic preparation of ginsenoside Rg3, which strain shows strong ability of converting ginsenoside Rb1and Rd into Rg3 in the solution with NaCl. The gene encoding a ${\beta}$-glucosidase from this GS514 was cloned and expressed in the BL21 (DE3) strain of Escherichia coli. The recombinant enzyme was purified and characterized. The molecular mass of purified was 87.5 kDa, as determined by SDS-PAGE. The gene sequence revealed significant homology to the family 3 glycoside hydrolases. The purified single enzyme also catalyzed the conversion of ginsenoside Rb1 into Rg3. This target enzyme will be able to produce as much saponin for consumer demand ginseng product. Anti-apoptotic proteins bind with pro-apoptotic proteins to induce apoptosis mechanism. Over expression of these anti-apoptotic proteins lead to several cancers by preventing apoptosis. Docking simulations were performed for anti-apoptotic proteins with several ginsenosides from Panax ginseng. Our finding shows ginsenosides particularly Rg3, Rh2 and Rf have more binding affinity with apoptotic proteins. Further, these docking system of each ginsenosides can be extended to experimental screen system for further brief confirmations of several diseases.

• Journal of Microbiology
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• 제43권2호
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• pp.152-157
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• 2005

Strain Kw07$^T$, a Gram-negative, non-spore-forming, rod-shaped bacterium, was isolated from granules in an Up-flow Anaerobic Sludge Blanket (UASB) bioreactor used in the treatment of brewery waste­water. 16S rRNA gene sequence analysis revealed that strain Kw07T belongs to the a-4 subclass of the Proteobacteria, and the highest degree of sequence similarity was determined to be to Sphingopyxis macrogoltabida IFO 15033T (97.8%). Chemotaxonomic data revealed that strain Kw07T possesses a quinone system with the predominant compound Q-I0, the predominant fatty acid C,s:, OJ7c, and sphingolipids, aU of which corroborated our assignment ofthe strain to the Sphingopyxis genus. The results of DNA-DNA hybridization and physiological and biochemical tests clearly demonstrated that strain Kw07T represents a distinct species. Based on these data, Kw07T (= KCTC 12209T = NBRC 100800T) should be classified as the type strain for a novel Sphingopyxis species, for which the name Sphingopyxis granuli sp. novo has been proposed.

• Journal of Microbiology and Biotechnology
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• 제18권1호
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• pp.110-117
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• 2008

Three kinds of prenylated flavonols, icariside I, icariside II, and icaritin, were isolated from an icariin hydrolysate and their effects on melanogenesis evaluated based on mushroom tyrosinase inhibition and quantifying the melanin contents in melanocytes. Although none of the compounds had an effect on tyrosinase activity, icariside II and icaritin both effectively inhibited the melanin contents with an $IC_{50}$ of 10.53 and $11.13{\mu}M$, respectively. Whereas icariside II was obtained from a reaction with ${\beta}$-glucosidase and cellulase, the icariin was not completely converted into icariside II. Thus, for the high-purity production of icariside II, the reaction was optimized using the response surface methodology, where an enzyme concentration of 5.0mg/ml, pH 7, $37.5^{\circ}C$, and 8 h reaction time were selected as the central conditions for the central composite design (CCD) for the enzymatic hydrolysis of icariin into icariside II using cellulase. Empirical models were developed to describe the relationships between the operating factors and the response (icariside II yield). A statistical analysis indicated that all four factors had a significant effect (p<0.01) on the icariside II production. The coefficient of determination $(R^2)$ was good for the model (0.9853), and the optimum production conditions for icariside II was an enzyme concentration of 7.5mg/ml, pH 5, $50^{\circ}C$, and 12 h reaction time. A good agreement between the predicted and experimental data under the designed optimal conditions confirmed the usefulness of the model. A laboratory pilot scale was also successful.

• 한국유기농업학회:학술대회논문집
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• 한국유기농학회 2009년도 하반기 학술대회
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• pp.286-286
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• 2009

벼 유기재배시 토양양분공급용으로 이용되고 있는 유기자재(금수강산골드)를 대조로 하고 식물성유기 자재(쌀겨팰렛), 동물성유기자재, 식물성과 동물성이 혼합된 유기자재를 질소 성분량(7kg/10a)을 기준으로 하여 이앙 20일전에 전량 기비로 시비하고 경운한 다음 동진1호를 시험품종으로 하여 2년연속 유기 자재와 벼를 재배하면서 일어나는 토양의 이화학적 특성과 벼 생육 및 특성의 변화를 시기별로 조사하였다. 시험 전 토양의 화학성은 전반적으로 유기물은 높고 인산함량은 매우 낮은 조건의 토양이었다. 관행유기자재(금수강산골드)는 20일경에 50% 무기화율을 보였으나, 식물성자재 40~60일경, 동물성자재와 혼합자재(식물성+동물성)는 60~80일경에 47~52% 무기화 정도를 나타내 식물성 자재의 무기화 속도가약 20일정도 빨랐다. 토양 중의 유기물 잔존함량은 식물성자재 > 혼합자재 > 동물성자재 > 관행 순이었으며, 토양 중의 전 질소 잔존함량의 경우 관행유기자재는 처리초기부터 빠르게 감소하는 특성을 보이나, 식물성자재와 혼합 자재는 시비초기와 거의 비슷한 수준을 유지하였고, 동물성 자재는 서서히 감소되는 경향이었다. 토양 물리성은 액상과 공극율 다소 증가되는 경향이었으며 식물성과 혼합유기자재처리구가 컸으며, 토양 유효 입단 형성력에 있어서도 유사한 경향이었다. 벼 수량 특성은 관행유기자재보다 1년차에는 3~9%의 낮았으나, 2년 연속처리를 할 경우 관행유기자재를 처리할 때와 동일한 생산성을 기대할 수 있었다. PME와 $\beta$-Glucosidase의 효소활성은 관행유기자재 < 식물성자재 < 동물성자재 < 혼합유기자재의 순으로 높은 경향을 볼 수 있었다.

• 한국유기농업학회:학술대회논문집
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• 한국유기농학회 2009년도 하반기 학술대회
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• pp.285-285
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• 2009

벼 유기재배에 있어서 녹비작물을 이용하여 화학비료를 대신하고 있으나 녹비를 이용하기 위해서는 월동 전에 파종하고 이듬해 벼 이앙 전에 토양에 환원을 해야 하는 번거로움이 있다. 따라서 벼 재배 직전에 유기자재를 이용하여 화학비료를 대신하고자 했을 경우 유기자재를 전층시비와 표층시비의 차이에 따른 토양중의 이화학적 특성과 벼의 수량특성의 변화를 구명하였다. 벼 유기재배시 토양양분공급용으로 이용되고 있는 유기자재 4종을 공시하여 유기자재의 질소 성분량(7kg/10a)을 기준으로 하여 이앙 20일전에 시비방법별로 전층시비와 표층시비 2처리로 구분하여 전량 기비시비하고 경운한 다음 동진1호를 시험품종으로 하여 2년 연속 시비처리와 벼를 재배하면서 일어나는 토양의 이화학적 특성과 벼 생육 및 특성의 변화를 시기별로 조사하였다. 시험 전 토양의 화학성은 표층시비구의 염류농도, 가리와 석회의 함량이 다소 높아서 염류농도가 전층 시비구 보다 높은 조건의 토양이었다. 유기자재별 무기화 정도는 전층시비보다 표층시비를 할 때 약 20~30일 정도 빨랐다. 토양 중의유기물 잔존함량은 시비방법간의 큰 차이는 없었으나 표층시비를 할 경우 후기로 갈수록 다소 증가되는 경향이었으나, 전질소 잔존함량은 감소되었다. 토양 액상과 공극율은 전층시비>표층시비였으며, 입단 형성력도 같은 경향이었다. 토양 효소활성은 PME의 활성은 유기자재를 전층처리하였을 때 촉진되었으며, $\beta$-Glucosidase의 활성은 전층보다 표층처리시 활성이 높았다. 시비방법에 따른 벼의 수량 특성은 시비방법별로는 표층시비를 할 경우 전층시비보다 4~7%의 높은 특성을 보였으며, 관행대비 1년차에는 3~9%의 낮았으나, 2년 연속처리를 할 경우 대조구와 비슷해 지는 경향이었다.

• 한국신재생에너지학회:학술대회논문집
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• 한국신재생에너지학회 2009년도 추계학술대회 논문집
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• pp.505-505
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• 2009

The object is to optimize the best condition of organosolv pretreatment process with sulfuric acid as a catalyst. As a material, Pitch pine (Pinus rigida) was ground and sieved through 40-mesh screen, and Celluclast and $\beta$-glucosidase were used as enzymes for enzymatic hydrolysis. Pretreatment processes were carried out in the minibomb, and 20 g of materials with 200 ml of 50% ethanol solution (v/v) with 1% sulfuric acid as a catalyst. Pretreatment temperature was varied from $150^{\circ}C$ to $190^{\circ}C$, and time was varied from 0 to 20 min. Then, residual materials were used for enzymatic hydrolysis. The best conditions were selected by estimating followed enzymatic hydrolysis rate and degradable rates after pretreatment process. The highest value of enzymatic hydrolysis rate was obtained as 55 - 60% at 160 and at $180^{\circ}C$, but the value decreased under more severe conditions. As the residual rates decreased under severe conditions, it infered that the decrease of sugar contents limits enzymatic hydrolysis rates. Combined with enzymatic hydrolysis rate, degradable rates and H-factors, the temperatures at $160^{\circ}C$ for 20 min and at $180^{\circ}C$ for 0 min were concluded as the optimized conditions where have the lowest H-factor value for considering energy input.

• 신재생에너지
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• 제6권4호
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• pp.6-14
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• 2010

Pretreatment of cellulosic biomass is necessary before enzymatic saccharification and fermentation. Extrusion is a well established process in food industries and it can be used as a physicochemical treatment method for cellulosic biomass. Aqueous ammonia soaking treatment at mild temperatures ranging from 60 to $80^{\circ}C$ for longer reaction times has been used to preserve most of the cellulose and hemicellulose in the biomass. The objective of this study was to evaluate the effect of extrusion treatment on aqueous ammonia soaking method. Extrusion was performed with miscanthus sample conditioned to 2mm of particle size and 20% of moisture content at $200^{\circ}C$ of barrel temperature and 175rpm of screw speed. And then aqueous ammonia soaking was performed with 15%(w/w) ammonia solution at $60^{\circ}C$ for 1, 2, 4, 8, 12 hours on the extruded and raw miscanthus samples respectively. In the combined extrusion-soaking treatment, most compositions removal occurred within 1~2 hours and on a basis of 1 hour soaking treatment values, cellulose was recovered about 85% and other compositions, including hemicellulose, are removed about 50% from extruded miscanthus sample. The combined extrusion-soaking treated and soaking only treated samples were subjected to enzymatic hydrolysis using cellulase and ${\beta}$-glucosidase. The enzymatic digestibility value of combined extrusion-2 hours soaking treated sample was comparable to 12 hours soaking only treated sample. It means that extrusion treatment can shorten the conventional long reaction time of aqueous ammonia soaking. The findings suggest that the combination of extrusion and soaking is a promising pretreatment method to solve both problems for no lignin removal of extrusion and long reaction time of aqueous ammonia soaking.