• Title/Summary/Keyword: laccase

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The effect of environmental condition to the mycelial browning of Lentinula edodes (Berkeley) Sing. during sawdust bag cultivation (환경조건이 표고톱밥배지의 갈변에 미치는 영향)

  • Kim, Young-Ho;Jhune, Chang-Sung;Park, Soo-Chul;You, Chang-Hyun;Sung, Jae-Mo;Kong, Won-Sik
    • Journal of Mushroom
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    • v.7 no.3
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    • pp.115-121
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    • 2009
  • Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes ) is getting increased because log cultivation is getting difficult to get oak logs. It is important to make mycelia browning on the substrate surface in sawdust cultivation. This browned surface plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. The period for mycelia browning is so long that the sawdust cultivation of Shiitake mushroom can not spread well into the mushroom farms. In this article we would like to discuss about the effect of environmental condition to the mycelial browning during sawdust bag cultivation for the To reduce the period required for browning of substrates, sawdust substrates was illuminated light with difference intensity. One hundred Lux light illumination was needed for producing normal yield of fruit body but fruit body yield was low and abnormally shaped fruit body was produced when cultured under the dark condition of incubation. Illumination over 200lux is necessary for the successful browning of substrates during incubation. Optimum incubation temperature for browning of substrates and fruiting was $25^{\circ}C$. The treatment of cotton plug with different size to identify the effect of aeration on the browning of substrates and fruiting showed rapid mycelial growth and reduced the periods for browning as the size of cotton plug was bigger. However, yield of fruit body was the highest at 16mm diameter cotton plug as compared to 20mm of that. $CO_2$ content in vessel of substrates was low as the size of cotton plug was bigger during incubation. $CO_2$ content during incubation of substrate was highest in periods between 8 week and 14 week after inoculation of shiitake when substrate was changed color into brown. $C_2H_4$ content in vessel with substrates was highest at 8mm diameter cotton plug and it was increased by order of 12, 16, 20, 0, 4 mm diameter cotton plug during substrate incubation. Sawdust substrate was soaked in cold water for different time to identify soaking effect of sawdust substrate on fruit body yield and activities of enzymes in these substrates were investigated. The fruit body yield was increased up to 40% by soaking substrates in comparison with unsoaked substrates. The soaked substrates showed 165, 175g/1,000ml at treatment of 4 and 15 hours, respectively. Cellulose activities in soaked substrates were not changed with soaking time, but activities of laccase, lignin degradation enzyme, were drastically increased up to 4 times in comparison with unsoaked substrates.

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The cultivate characteristics and the wood rotting ability and type of the Kuehneromyces mutabilis Sing. et A. H. Smith (무리우산버섯균의 배양적 특징과 목재부후 특성 파악)

  • Yun, Dae-Ryoung;Chai, Jyung-Ki
    • Journal of Mushroom
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    • v.2 no.4
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    • pp.192-199
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    • 2004
  • The mycelial mass of K. mutabilis greatly increased at pH 5.5~6.0 but decreased pH 6.0. The linear mycelial growth wsa mostly supported on sawdust of Quercus accutisima and the mycelial density wsa high on sawdust of Q. accutisima and corn cob. Much mycelial distribution could be showen in ray parenchyma cell and ray tracheid. Severe degradation of ray parenchyma cell was found but little degradation of ray tracheid cell was found. The dry weight loss wsa 5.9% after agar-block test. And the pH wsa acidified from 6.07 to 4.31 and hot water extractives was decreased after degradation of Q. serrata sawdust by K. mutabilis.

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Mycelial Culture Conditions of Lepista nuda and Extracellular Enzyme Activity (민자주방망이버섯(Lepista nuda) 균사체 배양조건 및 효소활성)

  • Kim Sang-Dae;Kim Ji-Hye;Kim Jong-Bong;Han Yeong-Hwang
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.164-167
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    • 2005
  • The culture condition and medium composition for the enhanced mycelial growth of Lepista nuda DGUM 26501 were investigated. The optimal temperature and pH for the mycelial growth were $24^{\circ}C$ and $7.0\~8.0$, respectively. The partial pressure of oxygen for the enhanced mycelial growth was more than $10\%\;O_2$. When Czapek-Dox medium was used as a minimal medium, manitol and xylitol were very good carbon sources. Organic nitrogen sources were better than inorganic ones for mycelial growth. As the nitrogen source tested, com steep liquor, soytone and protease peptone were the best as a source of organic nitrogen sources. When ammonium phosphate as phosphorus sources was used, the enhanced mycelial growth was shown. Nicotinic acid was proved to be the most appropriate source of vitamin. After the mycelia of L. nuda DGUM 26501 was cultivated at $24^{\circ}C$ for 10 days in LNM broth (pH 7.0), the activities of extracellular enzyme were determined. The specific activity of $\alpha-amylase$ was much higher than those of other enzymes. However, little or no enzyme activities of $\beta-glucosidase$, CMCase, laccase and lipase were found.

Trametes villosa Lignin Peroxidase (TvLiP): Genetic and Molecular Characterization

  • Carneiro, Rita Terezinha de Oliveira;Lopes, Maiza Alves;Silva, Marilia Lordelo Cardoso;Santos, Veronica da Silva;Souza, Volnei Brito de;Sousa, Aurizangela Oliveira de;Pirovani, Carlos Priminho;Koblitz, Maria Gabriela Bello;Benevides, Raquel Guimaraes;Goes-Neto, Aristoteles
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.179-188
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    • 2017
  • White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the $7^{th}$ day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

Comparison of in vitro digestibility and chemical composition among four crop straws treated by Pleurotus ostreatus

  • Nie, Haitao;Wang, Ziyu;You, Jihao;Zhu, Gang;Wang, Hengchang;Wang, Feng
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.1
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    • pp.24-34
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    • 2020
  • Objective: The effects of Pleurotus ostreatus on the feed utilization of broad bean stalks (BBS), rape straw (RS), paddy straw (PS), and corn stalk (CS) was examined. Methods: The four roughages were co-cultured with Pleurotus ostreatus. The chemical composition; enzyme activities of laccase, carboxymethylcellulase (CMCase) and xylanase; carbohydrate and protein fractions (based on The Cornell Net Carbohydrate and Protein System [CNCPS]) were assessed at different days after inoculation (7, 14, 21, 28 d) and un-inoculated roughages (control, 0 d). The digestibility of nutrient components and the gas production of roughage with various incubation times were monitored at 0, 2, 4, 6, 9, 12, 24, 36, 48, 60, and 72 h using an in vitro ruminal fermentation method. Results: A higher CMCase activity (0.1039 U/mL) and earlier time to peak (14 d) were detected in Pleurotus ostreatus cultured with CS (p<0.05). Significantly, the incubation length-dependent responses of cumulative gas production were observed from 24 to 72 hours post fermentation (p<0.05), and these incubation length-dependent effects on cumulative gas production of PS and CS appeared earlier (24 h) for PS and CS than those (48 h) for BBS and RS (p<0.05). The fast-degradable carbohydrate (CA) content for all four roughages significantly increased over time (p<0.05). Nonetheless, increased degradation efficiency for CA treated with Pleurotus ostreatus was detected at both 21 and 28 days of incubation (p<0.05). With the exception of PS (p<0.05), there were no significant difference among the roughages (p>0.05) in slowly-degradable carbohydrate (CB2) at different incubation times (p<0.05). Conclusion: Assessment of the alterations in chemical composition, CNCPS system fractions, and the fermentation kinetics after biological pretreatment may yield a valuable database for evaluating the biological pretreatment of Pleurotus ostreatus in ruminant feed.

Optimization of Bisphenol A Biodegradation by Trametes versicolor (Trametes versicolor에 의한 Bisphenol A 생분해의 최적조건)

  • Kang, Ae-Ri;Choi, Hyoung-Tae;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.37-42
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    • 2008
  • Optimal conditions for the biodegradation of endocrine-disrupting bisphenol A (BPA) were examined for the white rot fungus Trametes versicolor isolated in Korea. T. versicolor degraded 100% of 50 mg/L bisphenol A during 12 hr in yeast extract-malt extract-glucose (YMG) medium. When BPA was added to the 5-day preincubated fungal culture in YMG medium, all BPA was removed in 2 hr. T. versicolor could efficiently degrade BPA at $35^{\circ}C$, pH 6 in YMG medium. T. versicolor could more easily remove BPA of $1{\sim}25\;mg/L$ than that of higher concentrations ($50{\sim}100\;mg/L$) in YMG medium. T. versicolor degraded 100% of 50 mg/L BPA for 36 h in a minimal medium, which is lower degradation rate than that in YMG medium. Optimal conditions for BPA biodegradation in the minimal medium were similar to those in YMG medium. When BPA (50 mg/L) was added into domestic wastewater, it could be completely removed by T. versicolor. During the biodegradation of BPA by T. versicolor in YMG medium, its estrogenic activity decreased.

Analysis of Mycological Characteristics and Lignocellulose Degradation of Gyrodontium sacchari (헌구두솔버섯균의 균학적 특성 및 목질계 섬유소의 분해 특성 분석)

  • Park, In-Cheol;Seok, Soon-Ja;Kim, Jeong-Seon;Yoo, Jae-Hong;Ahn, Jae-Hyung
    • The Korean Journal of Mycology
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    • v.43 no.4
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    • pp.239-246
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    • 2015
  • Two fungal strains were isolated from rods of Quercus sp. (NAAS02335) and Pinus densiflora (NAAS05299) in Korea. These strains were identified as Gyrodontium sacchari by their morphological and mycological characteristics. The optimal growth temperature of NAAS02335 and NAAS05299 are $25^{\circ}C$ and $30^{\circ}C$, respectively. Production of cellulase, xylanase, and ligninase was tested on agar media supplemented dyes or substrates. Production of cellulase and xylanase of NAAS05299 was higher than those of NAAS02335, however ligninase activity of NAAS02335 was higher than that of NAAS05299. The activities of cellulase, xylanase, and amylase of strain NAAS05299 were estimated at 6.7~10.2 times higher than that of NAAS02335. Laccase activity was only estimated by strain NAAS02335. The lignocellulytic enzymes are induced by substrates such as rice straw, wooden chips of pine, oak, and poplar. The NAAS05299 was able to degrade filter paper completely after 4 weeks of culturing in liquid media containing a piece of filter paper at $28^{\circ}C$ with continuous shaking. NAAS05299 was able to degrade rice straw, pine chips, and oak chips after 4 months in solid culture, however NAAS02335 decomposed only rice straw among tested 4 kinds of biomass.

Influence of Temperature on the Bacterial Community in Substrate and Extracellular Enzyme Activity of Auricularia cornea

  • Zhang, Xiaoping;Zhang, Bo;Miao, Renyun;Zhou, Jie;Ye, Lei;Jia, Dinghong;Peng, Weihong;Yan, Lijuan;Zhang, Xiaoping;Tan, Wei;Li, Xiaolin
    • Mycobiology
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    • v.46 no.3
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    • pp.224-235
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    • 2018
  • Temperature is an important environmental factor that can greatly influence the cultivation of Auricularia cornea. In this study, lignin peroxidase, laccase, manganese peroxidase, and cellulose in A. cornea fruiting bodies were tested under five different temperatures ($20^{\circ}C$, $25^{\circ}C$, $30^{\circ}C$, $35^{\circ}C$, and $40^{\circ}C$) in three different culture periods (10 days, 20 days and 30 days). In addition, the V4 region of bacterial 16S rRNA genes in the substrate of A. cornea cultivated for 30 days at different temperatures were sequenced using next-generation sequencing technology to explore the structure and diversity of bacterial communities in the substrate. Temperature and culture days had a significant effect on the activities of the four enzymes, and changes in activity were not synchronized with changes in temperature and culture days. Overall, we obtained 487,694 sequences from 15 samples and assigned them to 16 bacterial phyla. Bacterial community composition and structure in the substrate changed when the temperature was above $35^{\circ}C$. The relative abundances of some bacteria were significantly affected by temperature. A total of 35 genera at five temperatures in the substrate were correlated, and 41 functional pathways were predicted in the study. Bacterial genes associated with the membrane transport pathway had the highest average abundance (16.16%), and this increased at $35^{\circ}C$ and $40^{\circ}C$. Generally, different temperatures had impacts on the physiological activity of A. cornea and the bacterial community in the substrate; therefore, the data presented herein should facilitate cultivation of A. cornea.

Metabolizing analysis according to the sawdust media of Pleurotus ostreatus (산겨릅나무와 옻나무 톱밥배지에서 재배한 느타리버섯의 함유성분 분석)

  • Shin, Yu-Su;Yang, Bo-Hyun;Kang, Bo-Yeon;Kim, Hyun-Soo;Lee, Ji-Hyun;Hong, Yoon-Pyo;Lee, Sang-Won;Lee, Chan-Jung;Kim, Seung-Yoo
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.194-197
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    • 2011
  • The transitivity of Chemical constituents by Pleurotus ostreatus cultivated in different raw sawdusts, which are Acer tegmentosum MAX, Rhus verniciflura, was investigated. The HPLC chromatography patterns on the chemical constituents of P. ostreatus showed the similar chromatography patterns in the different raw sawdusts and control sawdust. The unknown chemical constituents of P. ostreatus cultivated in the mixed medium added 10 %, 20% raw sawdusts, respectively, were increased. But the significance results in the mixed medium added 50% raw sawdusts were not showed. The chromatography patterns of mycelia grown in media added the 80% MeOH extracts of A. tegmentosum and R. verniciflura showed the similar patterns in comparison with control mycelia. In the results, the secondary metabolites of functional media were not degrade and changed to other derivatives compounds by P. ostreatus.

Mycelial Culture and Fruiting Analysis of Panellus edulis Strains Collected in Korea (Panellus edulis 수집 균주의 균사배양 및 자실체 특성 분석)

  • Woo, Sung-I;Ryoo, Rhim;Jang, Yeongseon;Park, Youngae;Jeong, Yeun Sug;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.281-294
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    • 2018
  • Molecular analysis using the internal transcribed spacer region sequences revealed that the strains used in this study, which were formerly identified as Panellus serotinus, are Panellus edullis. After Universal Fungal PCR Fingerprinting (UFPF) analysis, eight strains of P. edulis were divided into two groups. We conducted fundamental research on mycelial growth and sawdust cultivation to understand the cultural characteristics of eight wild P. edulis strains collected from Korean forests. All strains showed faster and denser mycelial growth on potato dextrose agar (PDA) than on other media (malt extract agar, Sabouraud dextrose agar). Optimal conditions for mycelial growth were: $20^{\circ}C$ on PDA, $25^{\circ}C$ on potato dextrose broth (PDB), and pH 5~8 on PDB at $25^{\circ}C$. Two strains (NIFoS 2407, 3993) were selected as excellent strains based on mycelial growth and density on PDA. NIFoS 2792 showed high cellulase activities on carboxymethyl cellulose (CMC) agar, and NIFoS 2387 and 2804 exhibited high laccase activities on ABTS-containing agar media. The mycelial growth of P. edulis was the fastest on Quercus acutissima and Q. mongolica sawdust media, and mycelial density was the highest on Quercus spp. sawdust-containing media. Sawdust cultivation of P. edulis was successful. The conditions were 80~85 days of cultivation period after spawn inoculation, 10~11 days for primordial formation at $17{\sim}18^{\circ}C$, and 15~20 days for fruiting growth. NIFoS 2804 and 3993 were selected as good strains in terms of cultivation period and mushroom production. These results could be useful for the artificial cultivation of P. edulis.