• 미생물학회지
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• 제46권1호
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• pp.93-95
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• 2010

리그닌 분해효소 군을 가진 백색부후균들은 다핵방향족 화합물을 포함하여 염료와 폭약 및 내분비장애 물질의 분해 등 다양한 난분해성 물질을 분해할 수 있다. 리그닌 분해효소 중 laccase와 manganese peroxidase (MnP)를 각각 발현하는 벡터를 국내에서 분리한 백색부후균류의 하나인 아교버섯(Phlebia tremellosa)에 형질전환 방법으로 도입시킨 형질전환체를 사용하여 염료의 탈색능력을 분석하였다. Methyl green의 경우 3일 후 약 50%의 탈색을 보인 야생형에 비하여 laccase 형질전환체(TF2-1)와 MnP 형질전환체(T5) 모두 90% 이상의 탈색을 보였다. Remazol brilliant blue R (RBBR)에서는 야생형이 약 67%의 탈색을 보인 반면 두 가지 형질전환체들은 약 85%의 탈색을 보였다. 각각의 형질전환체들은 laccase와 MnP의 활성 및 유전자 발현도 활발한 것으로 확인되었다.

• 한국버섯학회지
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• 제17권4호
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• pp.185-190
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• 2019

리그닌 분해 담자균류로 알려져 있는 느타리버섯균 No.42는 망간퍼옥시데이즈(MnP) 및 락게이즈(Lac)를 생산하였으나 글루코오스-펩톤-이스트-밀기울(GPYW)배지를 이용한 정치배양조건하에서 리그닌퍼옥시데이즈(LiP)활성은 검출되지 않았다. 한편, 동일배지에서 망간퍼옥시데이즈 활성은 11일째 80(3.6) U/flask(ml)의 최대 생산되었다. 망간퍼옥시데이즈 분리정제는 Sepha-ros CL-6B 및 Mono-Q 컬럼순으로 수행하였으며 주요 망간퍼옥시데이즈 isozyme은 단일밴드로 분자량은 36.4KDa이였다. N-말단으로부터 19개의 아미노산 배열은 단백질 자동 분석 장치로 분석한 결과 ATCADGRTTANAACCVLFP를 나타내었다. 느타리버섯균 No.42의 정치배양조건 하에서 세포외로 생산되는 중요한 망간퍼옥시데이즈 isozyme의 N-말단 아미노산 배열은 이전에 보고된 MnP3의 아미노산 배열과 동일하였다.

• 한국균학회지
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• 제31권3호
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• pp.181-186
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• 2003

담자균류 80여종에서 laccase 고생산 균주(Fomitella fraxinea; 장수버섯)를 선발하고, F. fraxinea를 이용한 리그닌 분해효소의 생산 및 효소의 대량생산 조건을 검토하였다. 먼저 배지 조성에 따른 생산성을 조사한 결과 Cu등의 무기물의 첨가 유무에 따라 생산되는 효소가 달랐다. 무기물이 첨가되지 않은 배지(Medium I)에서는 manganese peroxidase가 생산되었고, 무기물이 첨가된 배지(Medium II)에서는 laccase가 생산되었는데 플라스크 배양 18일째에는 5.43 U/ml, 20일째에는 5.56 U/ml의 효소활성이 나타났다. 배양방법에서 영양이 충분한 조건에서는 정치배양보다는 진탕배양(120 rpm)에서 효소의 활성이 높았다. 이 균주를 이용한 laccase 대량생산 실험에는 jar fermentor, balloon type bioreactor, air-sparging fermentor를 이용하였다. laccase는 jar fermentor와 air-sparging formentor에서 3,540 U/ml(8일), 3,100 U/ml(6일)의 생산성을 나타내었다. Balloon type bioreactor는 균사의 생장은 좋았지만 효소는 생산되지 않았다. F. fraxinea를 사용하여 air-sparging fermentor로 scale up하여 laccase의 대량생산을 할 수 있다고 판단된다.

• 한국버섯학회지
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• 제16권4호
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• pp.272-278
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• 2018

본 연구에서는 원목재배용 표고(Lentinula edodes) 품종에 대한 목질섬유소 분해능을 검정하였다. 5개의 국산품종(천백고, 산조 303호, 풍년고, 백화향, 수향고)을 대상으로 malt extract broth(MEB) 배지에 lignin의 첨가에 따른 RBBR(Remazol Brilliant Blue R) 탈색능을 조사한 결과, 산조 303호와 풍년고가 배양 5일째부터 우수한 분해능을 보였다. 섬유소 분해효소인 MnP와 laccase의 활성은 풍년고가 배양 7일째 MnP 활성이 2,809U/mg, laccase 활성이 2,230U/mg으로 나타났고, 산조 303호가 배양 11일째 MnP 활성이 2,673U/mg, laccase 활성이 2,049U/mg으로 최고 활성을 나타났으며, lignin을 첨가하였을 때 효소의 활성이 증가하였다. 산조 303호, 풍년고와 수향고는 배양 5주 만에 filter paper의 분해정도를 육안으로 확인이 가능할 정도로 분해능이 우수하였다.

• 한국버섯학회지
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• 제20권1호
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• pp.29-33
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• 2022

본 연구에서는 톱밥재배용 표고(Lentinula edodes) 품종에 대한 목질계 섬유소의 분해특성을 분석하였다. 국산 표고 품종 참아람, 산백향, 산조 713호, 산조 715호, 산조 718호를 대상으로 RBBR (Remazol Brilliant Blue R) 탈색능, 목질계 섬유소 분해 효소인 MnP와 laccase의 활성을 비교 분석하였다. 5개 품종의 탈색능과 효소활성은 품종별로 다른 양상을 보였다. 리그닌의 첨가에 따른 RBBR 탈색능을 조사한 결과, 참아람과 산조 713호가 각각 배양 3일과 5일째부터 우수한 분해능을 보였다. 섬유소 분해 효소인 MnP와 laccase의 활성은 산조 713호가 배양 2일째 MnP 활성이 1,213 U/mg, laccase 활성이 1,421 U/mg, 다음으로 참아람이 배양 7일째 MnP 활성이 1,921 U/mg, laccase 활성이 2,123 U/mg으로 최고 활성을 나타냈다. 이러한 결과는 표고 육종에 있어 리그닌 분해능이 우수한 균주의 선발이 가능함에 따라 교배균주의 육종 모본 활용에 효율적일 것으로 판단된다.

• 펄프종이기술
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• 제29권2호
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• pp.83-90
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• 1997

An unbleached hardwood kraft pulp was bleached in vitro with partially purified manganese peroxidase (MnP) from the fungus Phanerochaete sordida YK-624 without the addition of MnSO$_4$ in the presence of oxalate, malonate or gluconate known as manganese chelator, When the pulp was treated without the addition of MnSO$_4$, the pulp brightness increased by about 10 points in the presence of 2 mM oxalate, but the brightness did not significantly increase in the presence of 50 mM malonate. Residual MnP activity decreased faster during the bleaching with MnP without MnSO$_4$ in the presence of malonate than in the presence of oxalate. Oxalate reduced MnO$_2$ which already existed in the pulp or was produced from $Mn^{2+}$ by oxidation with MnP and thus supplied $Mn^{2+}$ to the MnP system. Thus, bleaching of hardwood kraft pulp with MnP, using manganese originally existing in the pulp, became possible in the presence of oxalate, a good manganese chelator and reducing reagent. Properties of partially purified MnPs from liquid cultures of white rot fungi, Ganoderma sp. YK-505, Phanerochaete sordida YK-624 and Phanerochaete chrysosporium were compared. MnP from Ganoderma sp. YK-505 was superior to MnPs from P. sordida YK-624 and P. chrysosporium in stabilities against high temperature and high concentration of $H_2O$$_2$. The MnP from Ganoderma sp. YK-505 differed in pH-activity profile from other MnPs. These data suggest that MnP from Ganoderma sp. YK-505 has different structure from those of other fungi. Bleaching of hardwood kraft pulp using the MnP from ganoderma sp. YK-505 is now in progress.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2014년도 춘계학술대회 및 임시총회
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• pp.43-43
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• 2014

White rot fungi have been useful source of enzymes for the degradation of environmental pollutants including polycyclic aromatic hydrocarbons (PAHs) and synthetic dyes. PAHs are widespread organic compounds present in fossil fuels and are routinely generated by incomplete fuel combustion. PAHs are some of the major toxic pollutants of water and soil environments. Synthetic dyes are major water-pollutants, which are toxic to organisms in water environments and interfere photosynthesis of water plants. Removal of PAHs and synthetic dyes has been of interests in the environmental science especially in the environmental microbiology. Mushrooms are fungal groups that function as primary degraders of wood polyphenolic lignin. The ligninolytic enzymes produced by mushroom, including manganese peroxidase, lignin peroxidase, and laccase, mediate the oxidative degradation of lignin. The catalytic power of these enzymes in the degradation of aromatic ring compounds has been sought for the degradation of various organic compounds. In this project, we have screened 60 wild mushroom strains for their degradation activity against two representative PAHs, naphthalene and anthracene, and five aromatic dyes, including alizarin red S, crystal violet, malachite green, methylene blue, rose bengal. The degradation of PAHs was measured by GC while the decolorization of dyes was measured by both UV spectrophotometer and HPLC. As results, 9 wild mushroom strains showed high activity in degradation of PAHs and textile dyes. We also describe the secretive enzyme activities, the transcription levels, and cloning of target genes. In conjunction with this, activities of degradative enzymes, including laccase, lignin peroxidase, and Mn peroxidase, were measured in the liquid medium in the presence of PAHs and dyes. Our results showed that the laccase activity was directed correlated with the degradation, indicating that the main enzyme acts on PAHs and dyes is the laccase. The laccase activity was further simulated by the addition of $Cu^{2+}$ ion. Detailed studies of the enzyme system should be sought for future applications.

• Journal of Ginseng Research
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• 제41권3호
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• pp.307-315
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• 2017

Background: Red-skin root disease has seriously decreased the quality and production of Panax ginseng (ginseng). Methods: To explore the disease's origin, comparative analysis was performed in different parts of the plant, particularly the epidermis, cortex, and/or fibrous roots of 5-yr-old healthy and diseased red-skin ginseng. The inorganic element composition, phenolic compound concentration, reactive oxidation system, antioxidant concentrations such as ascorbate and glutathione, activities of enzymes related to phenolic metabolism and oxidation, and antioxidative system particularly the ascorbate-glutathione cycle were examined using conventional methods. Results: Aluminum (Al), iron (Fe), magnesium, and phosphorus were increased, whereas manganese was unchanged and calcium was decreased in the epidermis and fibrous root of red-skin ginseng, which also contained higher levels of phenolic compounds, higher activities of the phenolic compound-synthesizing enzyme phenylalanine ammonia-lyase and the phenolic compound oxidation-related enzymes guaiacol peroxidase and polyphenoloxidase. As the substrate of guaiacol peroxidase, higher levels of $H_2O_2$ and correspondingly higher activities of superoxide dismutase and catalase were found in red-skin ginseng. Increased levels of ascorbate and glutathione; increased activities of $\text\tiny L$-galactose 1-dehydrogenase, ascorbate peroxidase, ascorbic acid oxidase, and glutathione reductase; and lower activities of dehydroascorbate reductase, monodehydroascorbate reductase, and glutathione peroxidase were found in red-skin ginseng. Glutathione-S-transferase activity remained constant. Conclusion: Hence, higher element accumulation, particularly Al and Fe, activated multiple enzymes related to accumulation of phenolic compounds and their oxidation. This might contribute to red-skin symptoms in ginseng. It is proposed that antioxidant and antioxidative enzymes, especially those involved in ascorbate-glutathione cycles, are activated to protect against phenolic compound oxidation.

• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.344-348
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• 2000

Abstract The removal percentage (94%) of 100 ppm of pyrene in a shaken culture of white rot fungus, Irpex lacteus, was much higher than that in a static culture (37.9%). Over 90% of the pyrene disappeared with I. lacteus grown at $15-27^{\circ}C$, yet less than 50% was removed at $37^{\circ}C$. The transformation rates of pyrene ($4.5-5.0{\;}\mu\textrm{g}/ml/day$) were not very different among cultures with 5- 30% inoculum sizes, and over 90% of the 100 ppm pyrene was removed in every case during 20 days of incubation. The biodegradation of pyrene by I. lacteus was confirmed by measuring the $CO_2$ evolved from the mineralization of the added pyrene. The activity of lignin peroxidase (LiP), which is known to be involved in the biodegradation by white rot fungi, was high between 8 to 12 days of incubation. Although manganese peroxidase activity was demonstrated during the same period as LiP, its activity was quite low, and no laccase activity was detected. Even though the activity patterns of ligninolytic enzymes did not coincide with the pyrene removal, this study shows that I. lacteus has a high biodegrading capability and can be a candidate for the bioremediation of polycyclic aromatic hydrocarbon contaminants.inants.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1147-1151
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• 2007

Biodegradation of endocrine-disrupting bisphenol A was investigated with several white rot fungi (Irpex lacteus, Trametes versicolor, Ganoderma lucidum, Polyporellus brumalis, Pleurotus eryngii, Schizophyllum commune) isolated in Korea and two transformants of T. versicolor (strains MrP 1 and MrP 13). I. lacteus degraded 99.4% of 50 mg/l bisphenol A in 3 h incubation and 100% in 12 h incubation. which was the highest degradation rate among the fungal strains tested. T. versicolor degraded 98.2% of 50 mg/l bisphenol A in 12 h incubation. Unexpectedly, the transformant of the Mn-repressed peroxidase gene of T. versicolor, strain MrP 1, degraded 76.5% of 50 mg/l bisphenol A in 12 h incubation, which was a lower degradation rate than wild-type T. versicolor. The removal of bisphenol A by I. lacteus occurred mainly by biodegradation rather than adsorption. Optimum carbon sources for biodegradation of bisphenol A by I. lacteus were glucose and starch, and optimum nitrogen sources were yeast extract and tryptone in a minimal salts medium; however, bisphenol A degradation was higher in nutrient-rich YMG medium than that in a minimal salts medium. The initial degradation of endocrine disruptors was accompanied by the activities of manganese peroxidase and laccase in the culture of I. lacteus.