• 유기물자원화
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• 제16권4호
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• pp.64-73
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• 2008

glysosyl hydrolase의 하나인 CCF 유사 유전자를 지렁이 Eisenia anderi의 중장으로부터 분리하여 클로닝하였다. 이 유전자의 전체 염기서열 크기는 1,152bp로 나타났으며 개시코돈을 포함하여 384개의 아미노산을 인코딩한다. N-말단 지역의 17개 잔기들은 signal peptide이다. CCF 관련 유전자의 아미노산 서열을 분석한 결과 본 연구에서 분리한 CCF 유사 유전자는 glycosyl hydrolase family 16 (GHF16)에 속하며, 다른 종의 지렁이에서 밝혀진 CCF 및 CCF-유사 단백질과 79~99%의 높은 상동성을 보였다. 여러 지렁이 종에서 분리된 CCF 및 CCF-유사 단백질들은 가수분해활성에 중요한 polysacchride-binding motif와 glucanase motif가 100% 상동성을 나타냈다. 본 연구의 대상종과 유사종인 E. fetida에서 분리된 CCF는 이 종의 서식지가 미생물 활성이 높은 부패 유기질층이기 때문에 다른 종의 CCF에 비해 높은 기질인식 특이성을 갖고 있는 것으로 알려져 있다. 이러한 사실은 본 연구에서 분리한 CCF도 넓은 기질 특이성을 갖고 있을 가능성을 제시해 주고 있으며 이는 산업적 응용 측면에서 유용한 특징 중의 하나라고 생각된다. BLASTX를 이용한 계통수 분석 결과 GHF16 효소는 크게 후생동물군, 녹색식물군, 진정세균군 등의 세 그룹으로 나눌 수 있었으며, 후생동물군의 GHF16은 다시 촉수담륜동물형와 탈피동물형(후생동물형 포함)으로 나뉘어졌다. 본 연구에 의해 E. andrei로부터 분리된 CCF-유사 단백질의 더 많은 생물학적 특성 연구를 통해 ${\beta}$-D-글루칸의 분해 및 생산을 조절하는 산업적 활용이 가능할 것으로 사료된다.

• Journal of the Korean Wood Science and Technology
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• 제47권6호
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• pp.692-699
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• 2019

Elizabethkingia miricola BM10, a symbiotic bacterium, has been isolated from the hindgut of Reticulitermes speratus KMT001, a termite which occurs on Bukhan Mountain in Seoul, Korea. This strain demonstrated a symbiotic characteristic, in that it lacked endo-${\beta}$-1,4-glucanase activity, in a previous study. The major fatty acids of E. miricola BM10 were iso-$C_{15:0}$, iso-$C_{17:0}$ 3-OH, and summed feature 3 (iso-$C_{16:1}{\omega}7c/C_{16:1}{\omega}6c$). The content of iso-$C_{17:0}$ 3-OH was higher, while those of ECL 13.566, iso-$C_{17:11}{\omega}9c$, and summed feature 4 were lower than the other three type-strains of the Elizabethkingia genus. The 16S rRNA phylogenetic analysis confirmed that E. miricola BM10 is a new species. The whole genome of E. miricola BM10 was sequenced. The average nucleotide identity of strain BM10 as evaluated by pairwise comparison with E. anophelis R26, E. meningoseptica ATCC 13253, and E. miricola GTC 862 was shown to be 91.5%, 81.2%, and 94.29%, respectively. Based on our study results, E. miricola BM10 appears to represent a new strain of the genus Elizabethkingia.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1196-1206
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• 2014

A metagenomic fosmid library was constructed using genomic DNA isolated from the gut microflora of Hermetia illucens, a black soldier fly. A cellulase-positive clone, with the CS10 gene, was identified by extensive Congo-red overlay screenings for cellulase activity from the fosmid library of 92,000 clones. The CS10 gene was composed of a 996 bp DNA sequence encoding the mature protein of 331 amino acids. The deduced amino acids of CS10 showed 72% sequence identity with the glycosyl hydrolase family 5 gene of Dysgonomonas mossii, displaying no significant sequence homology to already known cellulases. The purified CS10 protein presented a single band of cellulase activity with a molecular mass of approximately 40 kDa on the SDS-PAGE gel and zymogram. The purified CS10 protein exhibited optimal activity at $50^{\circ}C$ and pH 7.0, and the thermostability and pH stability of CS10 were preserved at the ranges of $20{\sim}50^{\circ}C$ and pH 4.0~10.0. CS10 exhibited little loss of cellulase activity against various chemical reagents such as 10% polar organic solvents, 1% non-ionic detergents, and 0.5 M denaturing agents. Moreover, the substrate specificity and the product patterns by thin-layer chromatography suggested that CS10 is an endo-${\beta}$-1,4-glucanase. From these biochemical properties of CS10, it is expected that the enzyme has the potential for application in industrial processes.

• The Plant Pathology Journal
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• 제30권4호
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• pp.355-366
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• 2014

In this study, resistance responses were investigated during the interaction of Botrytis fabae with two faba bean cultivars expressing different levels of resistance against this pathogen, Nubaria (resistant) and Giza 40 (susceptible). Disease severity was assessed on leaves using a rating scale from 1 to 9. Accumulation levels of reactive oxygen species (ROS), lipid peroxidation and antioxidant enzymes (superoxide dismutase, catalase and ascorbate peroxidase) were measured in leaf tissues at different times of infection. The expression profiles of two pathogenesis-related proteins (PRPs) encoded by the genes PR-1 and ${\beta}$-1,3-glucanase were also investigated using reverse transcription RT-PCR analysis. The accumulation of these defense responses was induced significantly in both cultivars upon infection with B. fabae compared with un-inoculated controls. The resistant cultivar showed weaker necrotic symptom expression, less ROS accumulation, a lower rate of lipid peroxidation and higher activity of the enzymatic ROS scavenging system compared with susceptible cultivar. Interestingly, ROS accumulated rapidly in the resistant leaf tissues and peaked during the early stages of infection, whereas accumulation was stronger and more intense in the susceptible tissues in later stages. Moreover, the response of the resistant cultivar to infection was earlier and stronger, exhibiting high transcript accumulation of the PR genes. These results indicated that the induction of oxidant/antioxidant responses and the accumulation of PRPs are part of the faba bean defense mechanism against the necrotrophic fungus B. fabae with a different intensity and timing of induction, depending on the resistance levels.

• The Plant Pathology Journal
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• 제31권2호
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• pp.195-201
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• 2015

Plant growth promoting rhizobacteria (PGPR) are known to confer disease resistance to plants. Bacillus sp. JS demonstrated antifungal activities against five fungal pathogens in in vitro assays. To verify whether the volatiles of Bacillus sp. JS confer disease resistance, tobacco leaves pre-treated with the volatiles were damaged by the fungal pathogen, Rhizoctonia solani and oomycete Phytophthora nicotianae. Pre-treated tobacco leaves had smaller lesion than the control plant leaves. In pathogenesis-related (PR) gene expression analysis, volatiles of Bacillus sp. JS caused the up-regulation of PR-2 encoding ${\beta}$-1,3-glucanase and acidic PR-3 encoding chitinase. Expression of acidic PR-4 encoding chitinase and acidic PR-9 encoding peroxidase increased gradually after exposure of the volatiles to Bacillus sp. JS. Basic PR-14 encoding lipid transfer protein was also increased. However, PR-1 genes, as markers of salicylic acid (SA) induced resistance, were not expressed. These results suggested that the volatiles of Bacillus sp. JS confer disease resistance against fungal and oomycete pathogens through PR genes expression.

• The Plant Pathology Journal
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• 제31권1호
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• pp.50-60
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• 2015

The use of novel isolates of Trichoderma with efficient antagonistic capacity against Fusarium oxysporum f. sp. lycopersici (FOL) is a promising alternative strategy to pesticides for tomato wilt management. We evaluated the antagonistic activity of 30 isolates of T. asperellum against 4 different isolates of FOL. The production of extracellular cell wall degrading enzymes of the antagonistic isolates was also measured. The random amplified polymorphic DNA (RAPD) method was applied to assess the genetic variability among the T. asperellum isolates. All of the T. asperellum isolates significantly reduced the mycelial growth of FOL isolates but the amount of growth reduction varied significantly as well. There was a correlation between the antagonistic capacity of T. asperellum isolates towards FOL and their lytic enzyme production. Isolates showing high levels of chitinase and ${\beta}$-1,3-glucanase activities strongly inhibited the growth of FOL isolates. RAPD analysis showed a high level of genetic variation among T. asperellum isolates. The UPGMA dendrogram revealed that T. asperellum isolates could not be grouped by their antagonistic behavior or lytic enzymes production. Six isolates of T. asperellum were highly antagonistic towards FOL and potentially could be used in commercial agriculture to control tomato wilt. Our results are consistent with the conclusion that understanding the genetic variation within Trichoderma isolates and their biochemical capabilities are required for the selection of effective indigenous fungal strains for the use as biocontrol agents.

• Asian-Australasian Journal of Animal Sciences
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• 제25권1호
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• pp.66-74
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• 2012

The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

• 한국미생물·생명공학회지
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• 제45권3호
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• pp.236-242
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• 2017

본 연구에서는 미역을 이용하여 초고온 열산 가수분해, 효소 당화, 발효과정을 거쳐 아세톤, 부탄올, 에탄올을 생성하는 실험에 대해 진행하였다. 초고온 열산 가수분해에서의 최적 조건은 10%의 slurry, 270 mM의 황산, $160^{\circ}C$에서의 7.5분이었다. 초고온 열산 가수분해는 열처리 시간을 줄이고 적은 농도의 황산을 사용해도 더 많은 당과 적은 저해물질을 생성해 낸다는 장점이 있다. 효소 당화에서는 Viscozyme L (${\beta}-glucanase$, Novozymes)을 12 unit/ml으로 처리하는 것이 25.1 g/l로 가장 많은 단당을 생성했다. 발효에서는 C. acetobutylicum KCTC 1724이 비교적 낮은 pH 5.0에서 많은 아세톤, 부탄올, 에탄올을 생성하는 장점이 있었지만 mannitol을 모두 소비하지 못하는 단점이 있어 고농도의 mannitol 배지에 순치한 C. acetobutylicum KCTC 1724을 사용하여 발효를 진행하였다. 그 결과, 아세톤, 부탄올, 에탄올이 각각 0.99 g/l, 5.62 g/l, 2.44 g/l로 순치하지 않은 C. acetobutylicum KCTC 1724를 이용해 발효했을 때 보다 부탄올은 2.45 g/l, 에탄올은 1.10 g/l 증가했으며 수율($Y_{ABE}$)은 0.24에서 0.37로 증가했다.

• 농업과학연구
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• 제44권4호
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• pp.525-530
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• 2017

Weaning is the most stressful event for nursery pigs because they are moved from familiar to unfamiliar environments. In addition, weaned pigs have immature digestive and immune systems. This situation makes weaned pigs susceptible to diseases and makes the absorption of nutrients from diets difficult. A feed approach, such as dietary enzyme supplementation, can be considered a solution. This study investigated the effects of dietary enzyme cocktail on diarrhea and immune responses of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.05% enzyme cocktail (Cocktail; combination of xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Incidence of diarrhea, packed cell volume (PCV), white blood cells (WBC) count, and immunoglobulin content were measured. A significantly lower incidence of diarrhea (p < 0.05) was observed in the Cocktail group as compared with the CON group. The Cocktail group also showed a decreased PCV (p < 0.1) on d 3 after weaning than the CON group. However, no differences were observed for number of WBC and contents of immunoglobulin G, M, and A between the Cocktail and CON groups. Consequently, inclusion of an enzyme cocktail in diets for weaned pigs had a positive influence on gut health by reducing the incidence of diarrhea in the present study.

• 대한지리학회지
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• 제39권4호
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• pp.643-664
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• 2004

본고는 지리학의 사회참여라는 주제 하에 한국의 지리학 전공자가 GIS 산업에 기여한 것과 수행한 역할을 가늠해보고자 시작되었다. 후학들의 기대수준과 실제적인 지리정보시스템을 비 지리학자의 요구사항과의 간격을 최소화하기 위하여, GIS 산업 현장에서 인정하고 있는 자격증 사안에 대한 소개와 향후 GIS산업의 방향을 예측하는 자료를 생산하는 것이 본고의 목적이다. 한국의 GIS 산업의 시기별 특성과 각 단계에 있어 지리학 전공자의 기여도 정리하였으며, 주요기관별 지리학 전공자의 비중 및 수행업무에 관한 인터뷰를 실시하였다. 또한 GIS 사업에 관련된 공공기관의 2003년 2004년의 중장기 기술개발 및 사업개발 계획을 분석하여 차후에 GIS 산업방향을 예측할 수 있도록 하였다. 설문지를 통하여 산업계와 연구계의 수요분석을 시행하고 그 결과는 GIS 산업의 진출을 위한 지리학과의 GIS교육의 SWOT (강점, 약점, 기회, 위협)분석으로 정리하였다. 분석결과 실습과 프로그래밍 중심의 훈련이 강조된 반면, 지리학과의 강점에 해당되는 다양한 세계 지리적 지식, 인간과 환경을 아우르는 종합적 시각. 통합능력, 학제간 연구훈련 등은 저평가되고 있었다. 대기업에서의 지리학과 전공자의 비중이 오히려 높았으며 이는 중소기업이 보다 구체적인 프로그램 구현부분에 기술적으로 치중하기 때문이다. 텔레매틱스와 같이 신성장동력으로 일컬어지는 산업에 GIS가 일부 포함되어 있기에. 지리학과의 GIS 교육은 문화지리와 연계된 지역마케팅, GIS 교육 분야, 새로운 위치기반의 서비스 모형개발 등에 기회가 있다고 여겨지며. 이는 학제간 협력 보다는 지리학내의 협동연구를 통한 핵심 역량 강화에 치중해야할 것을 시사한다. 보여준다.57장, 보조지 1장)이었으나, 수리 후 가로 25.8cm, 세로 39.4cm, 책의 두께는 1.9cm, 판본 64장(본문 57장, 보조지 1장, 겉표지 앞과 뒤 각1장, 보호지 앞과뒤 각2장)으로 가로 0.5cm, 세로 0.5cm, 두께 0.4cm, 판본 6장이 증가하는 등의 외형적 변화가 발생하였다.Polyacrylamide gel disc 전기(電氣) 영동(泳動)에서 L-c fraction은 pH 4. 3 gel에서는 명확(明確)한 단일(單一)의 단백질(蛋白質) 밴드를 보여주었으나, pH 8.3 gel에서는 아직도 두 개의 이상(以上)의 밴드가 검출(檢出)되었다. 4. 그러나 L-c fraction은 효모(酵母) 세포벽(細胞壁)의 다당류(多糖類) 성분중(成分中)에서 오직 glucan에 대한 ${\beta}-1$, 3-glucanase의 기질(基質) 특이성(特異性)만을 나타내었다. 따라서 L-c fraction을 zymolyase의 부분(部分) 정제(精製) 표품(標品)으로써 용해(溶解) 촉진(促進) 인자(因子)에 관(關)한 연구(硏究)에 제공(提供)하고져 동결(凍結) 건조(乾燥)하였다. used for court dress and official uniforms in the royal court. 4. Sa(紗) and Ra(羅) whose features have been examined we Gapsa(甲紗), Sun-in (純仁), Gosa(庫紗), Gwansa(官紗), Jusa(走紗), Eunjosa(은조사), Gwangsa(廣紗), Waesa(倭紗), Dorisa(도리사), Gong-yangsa(공양사), Rasa(羅紗), Danghangra(唐亢羅),