• Title/Summary/Keyword: laccase production

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Mycelial growth and wood decaying enzymatic activity analysis by various addition rates of oak powder in the liquid spawn of Lentinula edodes (참나무분 첨가에 따른 표고 액체종균의 균체생산 및 효소 활성)

  • Kim, Jeong-Han;Kang, Young-Ju;Baek, Il-Sun;Jeoung, Yun-Kyeoung;Lee, Yong-Seon;Cho, Hae-Seok;Lee, Young-Soon
    • Journal of Mushroom
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    • v.16 no.2
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    • pp.74-78
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    • 2018
  • This study was carried out to establish a suitable method for liquid spawn production from Lentinula edodes. The optimum production of liquid spawn (OLS) was achieved using soybean meal medium (SMM) with 0.3% of 850 um oak powder and 10-day incubation period and 0.6 vvm aeration volume. OLS showed activities of laccase on ABTS agar plate and carboxymethyl cellulase (CM-cellulase) on CMC agar plate. In case of liquid spawn, fruiting-body development period was delayed approximately 1 day compared to that of sawdust spawn, however, the yield of 153 g per 1.2 kg polypropylene bag was similar to that of sawdust spawn.

Degradation of Three Aromatic Dyes by White Rot Fungi and the Production of Ligninolytic Enzymes

  • Jayasinghe, Chandana;Imtiaj, Ahmed;Lee, Geon-Woo;Im, Kyung-Hoan;Hur, Hyun;Lee, Min-Woong;Yang, Hee-Sun;Lee, Tae-Soo
    • Mycobiology
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    • v.36 no.2
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    • pp.114-120
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    • 2008
  • This study was conducted to evaluate the degradation of aromatic dyes and the production of ligninolytic enzymes by 10 white rot fungi. The results of this study revealed that Pycnoporus cinnabarinus, Pleurotus pulmonarius, Ganoderma lucidum, Trametes suaveolens, Stereum ostrea and Fomes fomentarius have the ability to efficiently degrade congo red on solid media. However, malachite green inhibited the mycelial growth of these organisms. Therefore, they did not effectively decolorize malachite green on solid media. However, P. cinnabarinus and P. pulmonarius were able to effectively decolorize malachite green on solid media. T. suaveolens and F. rosea decolorized methylene blue more effectively than any of the other fungi evaluated in this study. In liquid culture, G. lucidum, P. cinnabarinus, Naematoloma fasciculare and Pycnoporus coccineus were found to have a greater ability to decolorize congo red. In addition, P. cinnabarinus, G lucidum and T. suaveolens decolorized methylene blue in liquid media more effectively than any of the other organisms evaluated in this study. Only F. fomentarius was able to decolorize malachite green in liquid media, and its ability to do so was limited. To investigate the production of ligninolytic enzymes in media containing aromatic compounds, fungi were cultured in naphthalene supple mented liquid media. P. coccineus, Coriolus versicolor and P. cinnabarinus were found to produce a large amount of laccase when grown in medium that contained napthalene.

Optimization of ${\beta}$-Glucosidase Production by a Strain of Stereum hirsutum and Its Application in Enzymatic Saccharification

  • Ramachandran, Priyadharshini;Nguyen, Ngoc-Phuong-Thao;Choi, Joon-Ho;Kang, Yun Chan;Jeya, Marimuthu;Lee, Jung-Kul
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.351-356
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    • 2013
  • A high ${\beta}$-glucosidase (BGL)-producing strain, Stereum hirsutum, was identified and isolated and showed a maximum BGL activity (10.4 U/ml) when cultured with Avicel and tryptone as the carbon and nitrogen sources, respectively. In comparison with other BGLs, BGL obtained from S. hirsutum showed a higher level of activity to cellobiose ($V_{max}$ = 172 U/mg, and $k_{cat}$ = 281/s). Under the optimum conditions (600 rpm, $30^{\circ}C$, and pH 6.0), the maximum BGL activity of 10.4 U/ml with the overall productivity of 74.5 U/l/h was observed. BGL production was scaled up from a laboratory scale (7-L fermenter) to a pilot scale (70-L fermenter). When S. hirsutum was cultured in fed-batch culture with rice straw as the carbon source in a 70-L fermenter, a comparable productivity of 78.6 U/l/h was obtained. Furthermore, S. hirsutum showed high levels of activity of other lignocellulases (cellobiohydrolase, endoglucanase, xylanase, and laccase) that are involved in the saccharification of biomasses. Application of S. hirsutum lignocellulases in the hydrolysis of Pinus densiflora and Catalpa ovata showed saccharification yields of 49.7% and 43.0%, respectively, which were higher than the yield obtained using commercial enzymes.

Production of Cellulase from Cellulomonas sp. KL-6 (Cellulomonas sp. KL-6에 의한 섬유소 분해효소의 생산)

  • Chung, Yung-Gun;Kwon, Oh-Jin
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.490-495
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    • 1995
  • Among the cellulases by Cellulomonas sp. KL-6. CMCase and filter paperase, which were produced as the out enzymes of cell, had been much produced, but very small amounts of ${\beta}-glucosidase $, the enzyme of which is cell bound form, was produced by this organism. The optimal culture times for CMCase and filter paperase productions were 5 days, while that of ${\beta}-glucosidase$ was 4 days. When this strain was cultured under the optimal medium for enzyme production, CMCase, FPase and ${\beta}-glucosidase$ were $82\;units/m{\ell},\;80\;units/m{\ell}\;and\;1.2\;units/m{\ell}$, respectively. Thus these results were showed to increase enzyme productivities as about $60{\sim}70%$ than those produced in basal medium. $CaCO_3$ injected to the medium as the ratio of 0.1% was not only enhanced cellulase activities but also effective as acid neutralizing agent. The production effects of lignase and lactase by this bacterium in filter paper medium was not appeared.

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Functional Characterization of Genes Located at the Aurofusarin Biosynthesis Gene Cluster in Gibberella zeae

  • Kim, Jung-Eun;Kim, Jin-Cheol;Jin, Jian-Ming;Yun, Sung-Hwan;Lee, Yin-Won
    • The Plant Pathology Journal
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    • v.24 no.1
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    • pp.8-16
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    • 2008
  • Aurofusarin is a polyketide pigment produced by some Fusarium species. The PKS12 and GIP1 genes, which encode a putative type I polyketide synthase (PKS) and a fungal laccase, respectively, are known to be required for aurofusarin biosynthesis in Gibberella zeae (anamorph: Fusarium graminearum). The ten additional genes, which are located within a 30 kb region of PKS12 and GIP1 and regulated by a putative transcription factor (GIP2), organize the aurofusarin biosynthetic cluster. To determine if they are essential for aurofusarin production in G. zeae, we have employed targeted gene deletion, complementation, and chemical analyses. GIP7, which encodes O-methyltransferase, is confirmed to be required for the conversion of norrubrofusarin to rubrofusarin, an intermediate of aurofusarin. GIP1-, GIP3-, and GIP8-deleted strains accumulated rubrofusarin, indicating those gene products are essential enzymes for the conversion of rubrofusarin to aurofusarin. Based on the phenotypic changes in the gene deletion strains examined, we propose a possible pathway for aurofusarin biosynthesis in G. zeae. Our results would provide important information for better understanding of naphthoquinone biosynthesis in other fdarnentous fungi as well as the aurofusarin biosynthesis in G. zeae.

Optimization of in Vitro Cultivation of Inonotus Obliquus

  • Cho, Nam-Seok;Shin, Yu-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.33 no.5 s.133
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    • pp.92-98
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    • 2005
  • This study was performed to get the basic information concerned to the optimum culture condition of Inonotus obliquus. Several solid media, PDA, MEA and Czapek-Dox, and three liquid media were adopted for the in vitro cultivation. Some main features of the fungal morphological characteristics under cultivation conditions were observed and described. Preliminary results showed that appearance of the mycelial mat, hyphal size and substrate pigmentation differed according to the media. The PDA medium was the most favorable substrate for the growth on solid culture, followed by MEA and Czapek-Dox media. Concerned to the addition of amino acids, 5 amino acids, such as alanine, alginine, isoleucine, leucine and threonine, enhanced to the mycelial growth. Isoleucine was shown the best fungal growth. An important morphological hyphal structure for the fungus, the setae, was found in abundance and diverse its shape and size. In liquid culture, fresh potato broth was the best growth stimulant of the fungus, followed by Malt extract and potato broth. Addition of yeast extract to the liquid media had improved the biomass, but not laccase production.

USE OF ENZYMES FOR MODIFICATION OF DISSOLVED AND COLLOIDAL SUBSTANCES IN PROCESS WATERS OF MECHANICAL PULPING

  • Johanna Buchert;Annikka Mustrnata;Peter Spetz;Rainer Ekman;Kari Luukko
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 1999.11b
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    • pp.115-119
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    • 1999
  • During mechanical pulp production and blcaching wood components, such as extractives, carbohydrates and lignin are dissolved and dispersed into the process waters. These components are called dissolved and colloidal substances(DCS). DCS can accumulate during water circulation and can in turn affect paper machine runnability and also the strength and optical properties of the paper. In this work DCS fraction origination from TMP process were treated with enzymes acting on triglycerides. glucomannans, and lignin and the effect of enzymatic treatments on the water composition as well as sheet properies were evaluated. Lipases were found to modify the chemical structure of the extractives resulting in more hydrophilic fibre surface and subsequent improvement in the sheet strength properties. Mannanase treatment, on the other hand, destabilized pitch. As a result, aggregation of pitch to the fibres was observed which in turn resulted in impaired strength properties. Laccase could effectively polymerize lignans and the reaction products seemed to be sorbed onto the fibres.

Fundamental Cultural Characteristics of Chlorociboria spp. Native to Korea (한국에 자생하는 녹청균류의 기초 배양 특성)

  • Jeon, Sung-Min;Ka, Kang-Hyeon;Wang, Eun-Jin;Ryoo, Rhim;Jang, Yeongseon
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.145-160
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    • 2018
  • Members of Chlorociboria are soft-rot ascomycetes that produce blue-green pigment. We investigated the growth characteristics of two Korean species of Chlorociboria, eight strains of Chlorociboria aeruginascens and Chlorociboria poutoensis, under various culture conditions (solid media, temperature, pH) and screened them for extracellular enzyme activity. Although the growth rate was slow, all tested strains of Chlorociboria spp. grew well on potato dextrose agar (PDA; 16.3~42.6 mm after 60 days) or Sabouraud dextrose agar (SDA), but not on malt extract agar (MEA). Compared with C. aeruginascens strains, C. poutoensis strains exhibited higher expression of blue-green pigments on both PDA and SDA media. The optimal temperature for mycelial growth was $20{\sim}25^{\circ}C$, and mycelial growth was lower at $30^{\circ}C$ than at $10^{\circ}C$. All strains tended to have increased mycelial growth as the incubation temperature increased in the range of 10 to $20^{\circ}C$. The optimal pH of potato dextrose broth (PDB) for mycelial growth varied according to the strain under static culture conditions. Maximum biomass production was obtained at pH 6.0 for NIFoS 579 ($114.3{\pm}5.1mg/60days$), but it maintained a stable pigment expression under a broad pH spectrum. The activities of both cellulase and laccase were observed in all tested strains of Chlorociboria spp. Enzyme activities of NIFoS 579 were remarkably higher than those of the other strains. From these results, we suggest that C. poutoensis NIFoS 579 is a potential candidate for use as a source of natural blue-green dye.

Isolation and Mycelial Cultivation Submerged of Phellinus sp. (Phellinus sp.의 분리 및 균사체의 액체배양)

  • Kang, Tae-Su;Lee, Dong-Gi;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.257-267
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    • 1997
  • Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.

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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.