• Journal of the Korean Wood Science and Technology
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• 제46권6호
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• pp.670-680
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• 2018

Acanthophysium sp. KMF001, a wood rotting fungus, produces a strong crude enzyme complex that efficiently produces simple sugars from wood. The transcriptomic analysis of Acanthophysium sp. KMF001 identified 14 genes for putative glycoside hydrolases. Among them, isotig01043 was expressed heterogeneously in Escherichia coli BL21(DE3), and the expressed protein exhibited an endo-${\beta}$-1,4-xylanase activity which showed the optimum reaction at pH 5.0 and $30^{\circ}C$. The enzyme kinetic values of $K_m$ and $V_{max}$ were 25.92 mg/ml and $0.628{\mu}mole/mg/ml$, respectively. The enzymatic characteristics of the expressed xylanase showed a typical fungal xylanase. However, the bioinformatics analysis suggested that the protein encoded by isotig01043 was a novel xylanase based on a low identity when it was compared with the closest protein in the NCBI database and a similar protein domain with GH16_fungal_Lam16A_glucanase, which had not been earlier suggested as a xylanase.

• 한국식품과학회지
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• 제39권3호
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• pp.276-282
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• 2007

마늘의 향미성분을 가급적 그대로 유지하고 착즙수율을 향상시킬 수 있는 착즙방법을 개발하고자 하였다. 착즙수율을 향상시키기 위하여 protopectinase와 복합요소(cellulase, pectinesterase 및 ${\beta}-glucanase$ 등이 혼합된 효소)를 마늘펄프로부터 1차로 착즙하고 남은 잔사에 적용하여 마늘주스의 수율증가율과 향미성분 변화를 측정하였다. 착즙 전 마늘펄프 무게에 대하여 효소를 0.04, 0.08 및 0.12%씩 가하여 30, 60, 90 및 120분간 각각 가수분해하였다. 마늘주스의 수율 증가는 최대수율에 도달할 때까지 효소의 첨가량 및 반응시간의 증가와 더불어 증가하였다. 0.12%의 protopectinase를 첨가하여 90분간 가수분해하여 착즙한 결과 수율은 13.8%가 증가하였다. 복합효소를 0.12% 첨가하여 90분간 가수분해 후 착즙한 경우에는 최대수율인 14.5%의 증가효과가 있었다. GC로 휘발성 향기성분을 분석한 결과 효소를 처리하지 않은 대조구(control)와 총 피크면적으로 계산하여 비교할 경우 감소하였다. 0.12%의 protopectinase를 가하여 60분간 분해 후 제조한 마늘 주스의 휘발성 향기성분 향량은 약 6%가 감소하였다. 마늘주스의 유리당 함량은 0.12%의 protopectinase를 가하여 60분간 분해후 제조한 마늘 주스가 대조구와 가장 유사한 함량을 나타내었다. 유리아미노산 함량은 사용한 효소의 종류에 영향을 크게 받지 않는 것으로 나타났다. 이상의 결과로부터 0.12%의 protopectinase를 마늘착즙잔사에 가하여 60분간 분해 후 착즙한 액을 마늘펄프로부터 직접 착즙한 액과 혼합하면 착즙수율을 최대로 증가시키고 향미 성분의 변화를 최소화시킬 수 있는 것으로 나타났다.

• 한국미생물·생명공학회지
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• 제40권3호
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• pp.190-196
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• 2012

식물생장 촉진 홀몬 auxin을 생산하는 균주를 선발하기 위해 경북 경산시 소재 고추경작지 근권토양으로 부터 739종의 일반호기성 균주와 urease 생산 균주 80종 및 광합성 균주 303종과 같이 총 1000여종 이상의 균주를 분리하였다. 이 균주들을 대상으로 Salkowski test를 실시한 결과, auxin을 생산하는 158종의 일반호기성 균주와 70종의 urease 생산 균주 및 228종의 광합성 균주를 선발할 수 있었으며, Holbrook test를 통해 또 다른 식물생장 촉진 호르몬인 gibberellin도 대부분의 균주에서 생산되는 것을 확인할 수 있었다. 선발된 균주 중 항진균 물질인 ${\beta}$-Glucanase와 siderophore를 생산하고 다양한 병원성 진균에 대해 길항 범위를 가지는 6가지 균주 BCB14, BCB17, C10, HA46, HA143, HJ5를 toothpicking 및 대치배양을 통해 최종 선발할 수 있었으며, 분류학적으로 동정한 결과 6종 모두 B. subtilis BCB14, B. methylotrophicus BCB17, B. methylotrophicus C10, B. sonorensis HA46, B. subtilis HA143, B. safensis HJ5로 확인되었다.

• Journal of Ginseng Research
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• 제43권3호
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• pp.408-420
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• 2019

Background: Ginseng (Panax ginseng Meyer) is an invaluable medicinal plant containing various bioactive metabolites (e.g., ginsenosides). Owing to its long cultivation period, ginseng is vulnerable to various biotic constraints. Biological control using endophytes is an important alternative to chemical control. Methods: In this study, endophytic Trichoderma citrinoviride PG87, isolated from mountain-cultivated ginseng, was evaluated for biocontrol activity against six major ginseng pathogens. T. citrinoviride exhibited antagonistic activity with mycoparasitism against all ginseng pathogens, with high endo-1,4-${\beta}$-D-glucanase activity. Results: T. citrinoviride inoculation significantly reduced the disease symptoms caused by Botrytis cinerea and Cylindrocarpon destructans and induced ginsenoside biosynthesis in ginseng plants. T. citrinoviride was formulated as dustable powder and granules. The formulated agents also exhibited significant biocontrol activity and induced ginsenosides production in the controlled environment and mountain area. Conclusion: Our results revealed that T. citrinoviride has great potential as a biological control agent and elicitor of ginsenoside production.

• 한국작물학회지
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• 제43권3호
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• pp.172-178
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• 1998

Freezing-resistant plants can survive subzero temperatures by withstanding extracellular ice formation. During cold acclimation, their leaves accumulate antifreeze proteins (AFPs) that are secreted into the apoplast and have the ability to modify the normal growth of ice crystals. Three barley, two wheat and two rye cultivars were grown under two different temperature regimes (20/16$^{\circ}C$ and 5/2$^{\circ}C$, day/night). Apoplastic proteins from winter cereals were separated by SDS-PAGE and detected with antisera to AFPs from winter rye. Apoplastic proteins accumulated to much higher levels in cold-acclimated (CA) leaves compared with nonacclimated (NA) ones in winter cereals. After cold acclimation, the protein concentration of apoplastic extracts increased significantly from 0.088 $mgmL^{-1}$ to 0.448 $mgmL^{-1}$, with about 5-fold increment. Also, the apoplastic protein content per gram leaf fresh weight in CA leaves ranged from 31 $\mu\textrm{g}$ $(gFW)^{-1}$ to 120 $\mu\textrm{g}$ $(gFW)^{-1}$ with an averaged value of 77 $\mu\textrm{g}$ $(gFW)^{-1}$, and coefficients of variation of 54.9%. The CA leaves in Musketeer (a Canadian winter rye cultivar) showed the greatest AFPs and antifreeze activity followed by 'Geurumil' (a Korean winter wheat cultivar), and 'Dongbori l' (Korean facultative barley cultivar). The proteins secreted into the wheat leaf apoplast at CA condition were more numerous than those observed in winter rye, where two $\beta$-1,3-glucanase-like proteins (GLPs), two chitinase-like proteins (CLPs) and two thaumatin-like proteins (TLPs) accumulated during cold acclimation. The proteins in barley leaf apoplast at CA conditions were a little different from those in wheat leaves. The AFPs were various among and within species. More freezing-resistant cultivars had more clear and numerous bands than less freezing-resistant ones. The high determination coefficient ($R^2$ =91 %) between freezing resistance and AFPs per gram leaf fresh weight indicated that the amount of AFPs was highly related to freezing resistance in winter cereal crops.

• 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.

• 미생물학회지
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• 제52권4호
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• pp.437-443
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• 2016

여러 토양에서 분리한 400여 개의 방선균 균주에 대해 4가지 식물병원성 진균에 대한 항진균 활성을 조사하였으며 그 중 Streptomyces costaricanus HR391 균주는 PDB 배지에서 식물병원성 진균인 Fusarium oxysporum f. sp. raphani, F. oxysporum f. sp. niveum, F. oxysporum f. sp. lycopersici와 Rhizoctonia solani의 균사 생장을대조군과 비교하여 각각 26.5, 26.2, 21.2와 23.8% 저해하였다. S. costaricanus HR391은 항진균물질인 siderophore를 $98{\mu}M$을 생성할 뿐만 아니라 유화활성을 나타내며 막지질을 파괴할 수 있는 생물계면활성제인 rhamnolipid와 lipopeptide인 iturin A와 surfactin를 생성하였다. 또한 진균 세포막을 분해할 수 있는 chitinase와 glucanase 활성도 나타내었으며 병원균의 막을 파괴하는 AMP와 항생물질 phenazine도 분비하였다. 이외에도 식물생장 촉진활성을 갖는 zeatin, gibberellin과 indole acetic acid 같은 식물호르몬도 생성하였다. 이와 같이 항진균 활성을 나타내는 S. costaricanus HR391 균주는 다양한 종류의 항진균 물질의 상승작용과 더불어 높은 생물계면활성이 본 균주의 항진균 활성에 큰 역할을 하는 것으로 보이며 친환경적인 생물학적 항진균제로서 활용이 가능할 것으로 기대된다.

• Asian-Australasian Journal of Animal Sciences
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• 제22권12호
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• pp.1676-1685
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• 2009

This trial was conducted to determine the effects of various feed additives on nutrient digestibility, performance and carcass traits of growing-finishing pigs fed diets containing wheat distiller' grains with solubles (WDGS). Seventy-two, individually fed pigs (19.7${\pm}$2.6 kg), were assigned to one of six dietary treatments in a 6${\times}$2 (treatment${\times}$sex) factorial design (N = 12). The control diet was based on wheat and soybean meal while the five experimental diets contained 20% WDGS during the growing period and 12% WDGS during the finishing period. One 20% WDGS diet was unsupplemented while the remaining diets were supplemented with either 0.1% threonine, 5% canola oil, 0.2% enzyme (0.1% Endofeed W containing 1,250 units/g of xylanase and 385 units/g of $\beta$-glucanase and 0.1% Vegpro containing 7,700 HUT/g protease and 75 CMC/g cellulase), or a combination of the three additives at the same levels as those fed separately. The digestibility of dry matter, crude protein and energy were all significantly higher in the control diet than the unsupplemented diet containing 20% WDGS. None of the feed additives improved nutrient digestibility. In addition, none of the additives had any significant effect on gain or feed intake during the growing (19.7 to 43.6) or finishing (43.6 to 114.3 kg) periods or overall (19.7 to 114.3 kg). During the growing period, feed conversion was significantly improved for pigs fed the combination of additives compared with the unsupplemented WDGS diet. During the finishing period and overall, feed conversion was significantly improved for pigs fed 5% canola oil alone or in combination with the other additives. None of the supplements had any effect on carcass traits. These results indicate that WDGS can be successfully used as a partial replacement for soybean meal in diets fed to growingfinishing pigs. However, due to its low energy content, there may be some merit in including high energy ingredients such as canola oil when diets containing WDGS are fed.

• 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.