• 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.

• 한국펄프종이공학회:학술대회논문집
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• 한국펄프종이공학회 2011년도 춘계학술발표회 논문집
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• pp.81-91
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• 2011

Current fuel ethanol research and development deals with process engineering trends for improving biotechnological production of ethanol. Recently, a large amount of studies regarding the utilization of lignocellulosic biomass as a good feedstock for producing fuel ethanol is being carried out worldwide. The plant biomass is mainly composed of cellulose, hemicellulose and lignin. The main challenge in the conversion of biomass into ethanol is the complex, rigid and harsh structures which require efficient process and cost effective to break down. The isolation of microorganisms is one of the means for obtaining enzymes with properties suitable for industrial applications. For these reasons, crude cultures containing cellulosic biomass degrading microorganisms were isolated from rice field soil, cow farm soil and rotten rice straw from cow farm. Carboxymethyl cellulose (CMC), xylan and Avicel (microcrystalline cellulose) degradation zone of clearance on agar platefrom rice field soil resulted approximately at 25 mm, 24 mm and 22 mm respectively. As for cow farm soil, CMC, xylan and Avicel degradation clearancezone on agar plate resulted around at 24mm, 23mm and 21 mm respectively. Rotten rice straw from cow farm also resulted for CMC, xylan and Avicel degradation zone almost at 24 mm, 23 mm and 22 mm respectively. The objective of this study is to isolatebiomass degrading microbial strains having good efficiency in cellulose hydrolysis and observed the effects of different substrates (CMC, xylan and Avicel) on the production of cellulase enzymes (endo-glucanase, exo-glucanase, cellobiase, xylanase and avicelase) for producing low cost biofuel from cellulosic materials.

• The Plant Pathology Journal
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• 제35권6호
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• pp.674-683
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• 2019

Some species of the Trichoderma genus are reported as the major problem in oak wood mushroom production in Korea. In spite of economic loss by the fungi, scientific information on airborne Trichoderma species is not much available. To generate information for disease management development we analyzed airborne Trichoderma. A total of 1,063 fungal isolates were purely obtained from indoor air sampling of cultivation houses used for oak wood mushroom using sawdust media. Among the obtained isolates, 248 isolates were identified as Trichoderma fungi including T. harzianum, T. atroviride, T. citrinoviride, and T. pseudokoningii, by morphological and molecular analysis. T. harzianum was dominant among the four identified species. All the four Trichoderma species grew fast on solid nutrient media tested (potato dextrose agar [PDA], malt extract agar [MEA], Czapek's Dox + yeast extract agar [CYA] and cornmeal dextrose agar). Compact mycelia growth and mass spore production were better on PDA and CYA. In addition, T. harzianum and T. citrinoviride formed greenish and yellowish mycelium and spores on PDA and CYA. Greenish and yellowish pigment was saturated into PDA only by T. pseudokoningii. These four Trichoderma species could produce extracellular enzymes of sawdust substrate degradation such as β-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease. Their mycelia inhibited the growth of oak wood mushroom mycelia of two tested cultivars on dual culture assay. Among of eleven antifungal agents tested, benomyl was the best to inhibit the growth of the four Trichoderma species. Our results demonstrate that the airborne Trichoderma fungi need to be properly managed in the cultivation houses for safe mushroom production.

• Asian-Australasian Journal of Animal Sciences
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• 제24권3호
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• pp.364-368
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• 2011

An experiment was conducted to examine the effects of Cordyceps militaris mycelia on microbial populations and fibrolytic enzyme activities in vitro. C. militaris mycelia was added to buffered rumen fluid with final concentrations of 0.00, 0.10, 0.15, 0.20, 0.25 and 0.30 g/L and incubation times were for 3, 6, 9, 12, 24, 36, 48 and 72 h. At all incubation times, the supplementation of C. militaris mycelia linearly increased the number of total viable and celluloytic bacteria; maximum responses were seen with 0.25 g/L supplementation of C. militaris mycelia. The addition of C. militaris mycelia above the level of 0.20 g/L significantly (p<0.01) increased the number of total and cellulolytic bacteria compared with the control. On the other hand, the response of fungal counts to the supplementation of C. militaris mycelia showed a linear decrease; the lowest response was seen with 0.30 g/L supplementation of C. militaris mycelia. It would seem that C. militaris mycelia possess a strong negative effect on rumen fungi since the lowest level of C. militaris mycelia supplementation markedly decreased fungal counts. Carboxylmethyl cellulase activities were linearly increased by the addition of C. militaris mycelia except at 3 and 9 h incubation times. At all incubation times, the supplementation of C. militaris mycelia linearly increased the activities of xylanase and avicelase. In conclusion, the supplementation of C. militaris mycelia to the culture of mixed rumen microorganisms showed a positive effect on cellulolytic bacteria and cellulolytic enzyme activities but a negative effect on fungi.

• 한국유기농업학회지
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• 제20권3호
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.

• 한국균학회지
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• 제10권2호
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• pp.67-73
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• 1982

2% CMC (Carboxymethyl cellulose)를 탄소원으로 하여 $28^{\circ}C$에서 8일간 배양한 도열병균 (Pyriculariaoryzae, $C-7^{+t}$)의 배양액을$(NH_4)_2SO_4$ 염석, Sephadex G-150 및 DEAE-Sephadex A-25 column chromatography를 거쳐 순화하였다. 순화결과 $F_1,\;F_2,\;F_3$ 3개의 CMCase $(C_x)$, 1개의 Avicelase $(C_1)$ 및 1개의 ${\beta}-glucosidase$를 얻었는데 그중 $C_1$ 효소의 $F_3$만을 골라 실험을 계속했다. 이 효소의 활성도는 pH 6.0 과 $40^{\circ}C$에서 가장 높았으며 $40^{\circ}C$까지 안정한 효소인 것을 알 수 있었다. 이 효소의 Km과 Vmax값은 각각 $2.8{\times}10^{-3}mM$, 5.9 mmoles/hour이었고, 분자량은 Sephadex G-150 column chromatography에 의해 약 80,000으로 나타났다. 약 7배로 순화된 이 효소의 순화정도를 polyacrylamide gel electrophoresis에 의해 검증한 결과 1개의band를 나타냈다.

• KSBB Journal
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• 제22권2호
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• pp.114-118
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• 2007

제주도 토양 샘플에서 분리한 amylase 생산균 15번 균주는 100 mL flask level와 10 L 생물반응기에서 T. inhamatum KSJ1의 FPpase, amylase 효소활성과 거의 비슷하였으며, flask level에서 두 균주의 xylanase, avicelase, CMCase, Gluco-amylase, $\beta$-amylase의 효소활성 및 음식물쓰레기의 당화능력도 모두 거의 비슷하였다. 두 균주를 YMEA 고체배지와 광학현미경으로 그 형태학적 특징을 관찰한 결과 T. inhamatum KSJ1이 녹색빛을 띠는 것과는 달리 amylase 생산균 15번 균주는 황색빛을 띠는 사상균으로 확인되었으며, 두 균주는 동일한 균은 아니지만 동일한 속으로 사료된다.

• Mycobiology
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• 제43권4호
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• pp.458-466
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• 2015

Oak tree death caused by symbiosis of an ambrosia beetle, Platypus koryoensis, and an ophiostomatoid filamentous fungus, Raffaelea quercus-mongolicae, has been a nationwide problem in Korea since 2004. In this study, we surveyed the yeast species associated with P. koryoensis to better understand the diversity of fungal associates of the beetle pest. In 2009, a total of 195 yeast isolates were sampled from larvae and adult beetles (female and male) of P. koryoensis in Cheonan, Goyang, and Paju; 8 species were identified by based on their morphological, biochemical and molecular analyses. Meyerozyma guilliermondii and Candida kashinagacola were found to be the two dominant species. Among the 8 species, Candida homilentoma was a newly recorded yeast species in Korea, and thus, its mycological characteristics were described. The P. koryoensis symbiont R. quercusmongolicae did not show extracelluar CM-cellulase, xylanase and avicelase activity that are responsible for degradation of wood structure; however, C. kashinagacola and M. guilliermondii did show the three extracellular enzymatic activities. Extracelluar CM-cellulase activity was also found in Ambrosiozyma sp., C. homilentoma, C. kashinagacola, and Candida sp. Extracelluar pectinase activity was detected in Ambrosiozyma sp., C. homilentoma, Candida sp., and M. guilliermondii. All the 8 yeast species displayed compatible relationships with R. quercus-mongolicae when they were co-cultivated on yeast extract-malt extract plates. Overall, our results demonstrated that P. koryoensis carries the yeast species as a symbiotic fungal associate. This is first report of yeast diversity associated with P. koryoensis.

• 미생물학회지
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• 제36권3호
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• pp.181-186
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• 2000

Clostridium thermocellum이 생성하는 섬유소분해 효소복합체인 cellulosome은 26개 의 서로 다른 단백질로 구성되어 있으며 그 구성물질로서 calcium을 포함하고 있다. 견고한 구조의 이 복합체에서 구성단백질을 분리하여 그 기능을 연구할 목적으로 이 복합체를 해체 (dissociation)하려 시도하였다. 이 복합체는 calcium을 제거하였을 대 해체되었다. 해체된 구성 단백질들은 MonoQ column chromatogrphy에 의해 구조단백질인 CipA를 포함한 분획, 91 kDa(CelK-tr), 60 kDa 과 57 kDa 단백질로 구성된 분획과 주로 46 kDa(CelA-tr), 또는 71 kDa(CelS-tr) 단백질을 포함하는 분획들로 크게 분리되었다. 대부분의 분획들은 crystalline cellulose 분해 활성을 보였다. 순수 분리된 71 kDa 단백질은 $60^{\circ}C$~$70^{\circ}C$에서 섬 유소 분해시 calcium에 의존적이었으나 46 kDa 단백질은 그렇지 않았다. 46 kDa 단백질은 cellodextrin을 celloviose 및 cellotriose 단위로 절단하며 cellotetraose 로부터 glucose가 생 성되는 것이 관찰되었다. Cellulosome의 섬유소분해 최성 산물이 cellobiose인 것을 고려할 때 개개 구성단백질의 활성이 이 효소 복합체내에서 조절되어 있음을 알 수 있다.

• 정신신체의학
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• 제5권2호
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• pp.185-194
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• 1997

본 연구는 정신과외래에 내원한 52명의 신체형장애, 52명의 우울장애, 51명의 정신신체장애환자들을 대상으로 질병행동을 비교하기 위해 실시되었다 질병행동을 평가하기 위해서 질병행동평가스케줄(illness behavior assessment schedule) 및 치료추구행태에 관한 설문을 사용하였으며 면담방법은 구조적인 형태로 이루어졌다. 신체형장애환자들과 정신신체장애환자들은 각각 우울장애환자들에 비해 신체적 질병에 대한 확신, 질병에 대한 공포, 질병에 대한 집착이 더 강하고 한의원의 이용빈도가 더 높았다. 신체형장애환자들은 우울장애환자들에 비해 질병의 원인을 신체적인 것으로 더 확신하고 정동장애의 원인을 심리적인 것으로 귀착시키는 경향이 낮으며 우울 및 안절부절못하는 정도 및 정신과에 대한 수용도가 각각 더 낮았다. 한편 신체형장애환자들은 정신신체장애환자들보다 질병의 정도가 더 경한 것으로 설명되었음을 확신하였다. 신체형장애환자들은 정신적 고통이 있는 경우에 감정표현의 억제가 더 현저하였다. 신체형장애환자들 중 여자들이 남자들보다 더 심리적 장애가 있고 병의 원인을 심리적인 인자로 귀착시켜 여자에 대한 정신과적 접근이 더 용이할 수 있음을 시사하였다. 이상의 결과들은 질병 행동에 있어서 신체형장애환자들이 우울장애환자들과는 다른 반면 증상의 평가에 있어서 치료자와 환자간의 차이를 제외하고는 정신신체장애환자들과 비슷한 양상임을 시사한다. 따라서 신체형장애환자들에 대한 접근은 이런 환자들의 심리적인 것을 부정하려는 욕구를 이해하면서 처음에는 신체적인 접근으로 시작하여 점진적으로 심리적 및 생물정신사회적인 접근으로 전환해야 할 필요성이 강조된다.조절되어진다는 사실을 제시한다. 하지만 태생어류에서 이 호르몬의 실질적 작용 메카니즘 및 명확한 작용시기에 대해서는 보다 많은 연구가 요구되어 진다.을 설계하는 것이 가능하였다.적(最適) 온도(溫度)는 $30^{\circ}C$, avicelase와 ${\beta}-glucosidase$의 최적(最適) pH는 5.0, CMCase는 pH 5.5 이었으며, 균사(菌絲) 생육(生育)은 pH 5.0에서 양호(良好)하였다. 배양(培養) 기간(期間)은 avicelase가 8일(日), CMCase가 10일(日), ${\beta}-glucosidase$는 16 일간(日間) 배양(培養)하였을 때 최대치(最大値)를 보였고, 균사(菌絲) 생육(生育)은 12일(日) 배양(培養)했을 때 가장 양호(良好)하였다.가한 반면, 중국인들은 고소한 향의 강도, 고소한 향의 기호도, 전체적인 맛에서 뚜렷하게 일본참기름을 우수하게 평가하였다.s의 항체(抗體)로 반응(反應)시킨 후 protein-A gold(15 nm)로 표식(標識)시킨 바 제일 바깥 상층(上層)의 keratinocyte에 있어서 세포막표면(細胞膜表面)을 따라 표식(標識)되어 세포막항원(細胞膜抗元)을 나타내었으며, 이와 같은 소견(所見)으로 미루어 정상피부(正常皮膚) 중층편평상피세포(重層扁平上皮細胞)에서도 동일(同一)한 소견(所見)을 관찰(觀察)할 수 있다고 본다.al remnants, Resorption of fetus로 관찰된 것이다. Fetal death는 수정후 $14{\sim}18$일까지의 사망으로써 Maceration of fetus로