• 한국미생물·생명공학회지
• /
• 제48권4호
• /
• pp.463-470
• /
• 2020

삼림지역의 고목에서 분리한 JS18 균주는 합성 멜라닌을 탈색하는 세포 외 분비효소를 생산했다. JS18 균주의 internal transcribed spacer (ITS) 염기서열을 분석하고 계통학적으로 확인한 결과 본 균주는 Trametes velutina로 동정되었다. JS18 균주는 laccase 활성을 나타냈지만 manganese peroxidase 및 lignin peroxidase 활성은 나타내지 않았다. 본 균주를 회분배양한 결과 멜라닌 탈색 활성은 laccase 활성으로부터 유래하는 것으로 확인되었다. Laccase 유도인자로써 Syringic acid 및 CuSO4를 첨가하고 25℃에서 7일간 배양한 결과 배양상등액에서 98 U/ml의 laccase 활성을 나타내었다. GYP 배지에서 배양한 T. velutina의 배양상등액에서 ammonium sulfate 침전, Hi-trap Q Sepharose 컬럼 및 gel filtration을 이용하여 효소를 정제하였고, SDS-PAGE에서 약 67 kDa의 분자량을 나타내었다. 정제된 효소의 멜라닌 탈색율은 효소 단독으로는 24 시간 만에 단지 4% 만을 나타내는 반면, HBT의 존재 하에서는 80%로 향상되었다. 또한 1.5 mM HBT의 농도에서는 최대 81%의 멜라닌 탈색율을 나타내었다. 본 효소의 멜라닌 탈색에 대한 최적 pH 및 온도는 각각 5.0와 37℃를 나타내었다. 본 연구에서는 T. velutina JS18 유래 laccase가 촉매하는 멜라닌 탈색 반응에서 redox mediator로써 HBT의 적용 가능성을 확인하였다.

• 미생물학회지
• /
• 제46권1호
• /
• pp.93-95
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제31권6호
• /
• pp.823-832
• /
• 2021

Mushroom cultivation along with the palm oil industry in Malaysia have contributed to large volumes of accumulated lignocellulosic residues that cause serious environmental pollution when these agroresidues are burned. In this study, we illustrated the utilization of lignocellulolytic enzymes from the spent mushroom substrate of Pleurotus pulmonarius for the hydrolysis of palm oil mill effluent (POME). The hydrolysate was used for the production of biohydrogen gas and enzyme assays were carried out to determine the productivities/activities of lignin peroxidase, laccase, xylanase, endoglucanase and β-glucosidase in spent mushroom substrate. Further, the enzyme cocktails were concentrated for the hydrolysis of POME. Central composite design of response surface methodology was performed to examine the effects of enzyme loading, incubation time and pH on the reducing sugar yield. Productivities of the enzymes for xylanase, laccase, endoglucanase, lignin peroxidase and β-glucosidase were 2.3, 4.1, 14.6, 214.1, and 915.4 U g^-1, respectively. A maximum of 3.75 g/lof reducing sugar was obtained under optimized conditions of 15 h incubation time with 10% enzyme loading (v/v) at a pH of 4.8, which was consistent with the predicted reducing sugar concentration (3.76 g/l). The biohydrogen cumulative volume (302.78 ml H₂.L^-1 POME) and 83.52% biohydrogen gas were recorded using batch fermentation which indicated that the enzymes of spent mushroom substrate can be utilized for hydrolysis of POME.

• 한국버섯학회지
• /
• 제20권1호
• /
• pp.29-33
• /
• 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.

• 한국초지조사료학회지
• /
• 제34권1호
• /
• pp.15-20
• /
• 2014

To investigate lignification process and its physiological significance under water-deficit condition, the responses of peroxidases, polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) in relation to leaf water status to the short term of water deficit treatment in the leaves with different maturities in forage rape were measured. The significant decrease in relative water content (RWC) and leaf osmotic potential (${\Psi}{\pi}$) were apparent after 5 d of water-deficit treatment. The activity of guaiacol peroxidase (GPOD), ascorbate peroxidase (APOD), coniferyl alcohol peroxidase (CPOD), and syringaldazine peroxidase (SPOD) was depressed especially in middle and old leaves when compared with that of control leaves. On the other hand, in young leaves, a significant increase in CPOD (+34%) and SPOD (+24%) activity as affected by water-deficit treatment was apparent. The activation of PAL and PPO was observed in middle and old leaves for PAL and in young and middle leaves for PPO. These results suggest that peroxidases in middle and old leaves did not involve in lignification under mild water-deficit stress, whereas CPOD and SPOD in young leaves participate in lignification by a coordination with PAL and PPO to incorporate phenol and lignin into the cell walls.

• 미생물학회지
• /
• 제55권2호
• /
• pp.177-179
• /
• 2019

미국 뉴저지주 중위도 산림토양에서 부식산(천연 복합유기화합물인 부식질의 주요 구성성분) 분해능이 있는 세균 균주 Pseudomonas kribbensis CHA-19를 분리하였으며, 이후 또 다른 토양 유기물인 리그닌과 리그닌 유래의 페룰산(ferulic acid)과 바릴린산(vanillic acid)의 분해능을 확인하였다. 부식질 초기 저분자화 효소(예, dye-decolorizing peroxidase와 laccase-like multicopper oxidase)와 부식질 유래의 다양한 저분자 분해산물들을 분해하는 효소(예, vanillate O-demethylase와 biphenyl 2,3-dioxygenase)를 탐색하기 위해 CHA-19 게놈염기서열을 분석하였다. 최종 확보한 효소유전자 정보는 토양세균의 부식질 분해경로 제안에 사용되었다.

• Mycobiology
• /
• 제37권1호
• /
• pp.53-61
• /
• 2009

The process of biodegradation in lingo-cellulosic materials is critically relevant to biospheric carbon. The study of this natural process has largely involved laboratory investigations, focused primarily on the biodegradation and recycling of agricultural by-products, generally using basidiomycetes species. In order to collect super white rot fungi and evaluate its ability to degrade lingo-cellulosic material, 35 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye. In the laccase enzymatic analysis chemical test, 33 white rot fungi and 2 brown rot fungi were identified. The degradation ability of polycyclic aromatic hydrocarbons (PAHs) according to the utilized environmental conditions was higher in the mushrooms grown in dead trees and fallen leaves than in the mushrooms grown in humus soil and livestock manure. Using Poly-R 478 dye to assess the PAH-degradation activity of the identified strains, four strains, including Agrocybe pediades, were selected. The activities of laccase, MnP, and Lip of the four strains with PAH-degrading ability were highest in Pleurotus incarnates. 87 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye on solid media. Using Poly-R 478 dye to assess the PAHdegrading activity of the identified strains, it was determined that MKACC 51632 and 52492 strains evidenced superior activity in static and shaken liquid cultures. Subsequent screening on plates containing the polymeric dye poly R-478, the decolorization of which is correlated with lignin degradation, resulted in the selection of a strain of Coriolus versicolor, MKACC52492, for further study, primarily due to its rapid growth rate and profound ability to decolorize poly R-478 on solid media. Considering our findings using Poly-R 478 dye to evaluate the PAH-degrading activity of the identified strains, Coriolus versicolor, MKACC 52492 was selected as a favorable strain. Coriolus versicolor, which was collected from Mt. Yeogi in Suwon, was studied for the production of the lignin-modifying enzymes laccase, manganese-dependent peroxidase (MnP), and lignin peroxidase (LiP).

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

• 미생물학회지
• /
• 제55권1호
• /
• pp.83-85
• /
• 2019

남극 연안 툰드라 토양에서 리그닌 분해능이 있는 Pseudomonas sp. PAMC 29040를 분리하였으며, 이후 토양 유기물의 주요 구성성분인 복합유기화합물 부식질 분해능을 확인하였다. 부식질 초기 저분자화 효소(예, dye-decolorizing peroxidase)와 부식질 유래의 다양한 저분자 분해산물들을 분해하는 효소들(예, vanillate O-demethylase)를 탐색하기 위해 PAMC 29040 게놈 염기서열을 분석하였다. 분석을 통해서 최종 확보한 효소유전자 정보는 저온환경에 서식하는 토양 세균의 부식질 분해경로 제안에 활용될 것이다.

• Journal of the Korean Wood Science and Technology
• /
• 제35권2호
• /
• pp.85-95
• /
• 2007

Manganese dependent peroxidase (MnP) is the most ubiquitous enzyme produced by white-rot fungi, MnP is known to be involved in lignin degradation, biobleaching and oxidation of hazardous organopollutants. Bjerkandera fumosa is a nitrogen-unregulated white-rot fungus, which produces high amounts of MnP in the excess of N-nutrients due to increased biomass yield. The objective of this study was to optimize the MnP production in N-sufficient cultures by varying different physiological factors such as Mn concentration, culture pH, and incubation temperature. The growth of fungus was optimal in pH 4.5 at $30^{\circ}C$, $N_2$-unregulated white-rot fungus produces high amounts of MnP in the excess N-nutrients. The fungus produced the highest level of MnP (up to $1000U/{\ell}$) with $0.25g/{\ell}$ asparagine and $1g/{\ell}$ $NH_4Cl$ as N source at 1.5 mM $MnCl_2$ concentration, pH value of 4.5 at $30^{\circ}C$. Purification of MnP revealed the existence of two isoforms: MnPl and MnP2. The molecular masses of the purified MnPl and MnP2 were in the same range of 42~45 kDa. These isoforms of B. fumosa strictly require Mn to oxidize phenolic substrates. Concerned to kinetic constants of B. fumosa MnPs, B. fumosa has similar Km value and Vmax compared to the other white-rot fungi.