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Yasushi Sugano - One of the best experts on this subject based on the ideXlab platform.
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Purification and characterization of two DyP isozymes from Thanatephorus cucumeris Dec 1 specifically expressed in an air-membrane surface bioreactor.
Journal of bioscience and bioengineering, 2009Co-Authors: Takuya Shimokawa, Makoto Shoda, Yasushi SuganoAbstract:DyP isozymes (DyP2 and DyP3) from the culture fluid of the fungus Thanatephorus cucumeris Dec 1 by air-membrane surface bioreactor were purified and characterized. The characteristics of DyP2 were almost the same as those of a recombinant DyP reported previously, but different from DyP3.
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Efficient dye decolorization and production of dye decolorizing enzymes by the basidiomycete Thanatephorus cucumeris Dec 1 in a liquid and solid hybrid culture.
Journal of bioscience and bioengineering, 2008Co-Authors: Takuya Shimokawa, Makoto Shoda, Mitsuyo Hirai, Yasushi SuganoAbstract:Thanatephorus cucumeris Dec 1, a basidiomycete, is a promising decomposer of several xenobiotics. Air-membrane surface bioreactor culture (AMS culture), a hybrid between solid-state and submerged culture, was used for Dec 1 growth. The optimum temperature for AMS culture (25°C) was lower than that for submerged culture (30°C). Fungal growth was almost the same in AMS culture as in submerged culture at optimum temperatures. However, protein secretion, dye decolorizing peroxidase (DyP) activity, and aryl alcohol oxidase activity, were 18-, 233-, and 36.5-fold higher in AMS culture than in submerged culture, respectively, indicating that AMS culture of Dec 1 was superior to submerged culture. In in vivo dye decolorization tests, 13 of 16 dyes were decolorized by more than 90% within 10 days. Interestingly, under AMS culture, a biofilm was formed; biofilm formation was not, however, essential for DyP and manganese dependent peroxidase (MnP) activities. Although the correlation between DyP activity and water activity was unclear, MnP activity seemed to increase in activity with decreasing water activity, even when no biofilm formation was observed.
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Complete decolorization of the anthraquinone dye Reactive blue 5 by the concerted action of two peroxidases from Thanatephorus cucumeris Dec 1
Applied microbiology and biotechnology, 2006Co-Authors: Yasushi Sugano, Yuko Matsushima, Makoto ShodaAbstract:It is useful to identify and examine organisms that may prove useful for the treatment of dye-contaminated wastewater. Here, we report the purification and characterization of a new versatile peroxidase (VP) from the decolorizing microbe, Thanatephorus cucumeris Dec 1 (TcVP1). The purified TcVP1 after Mono P column chromatography showed a single band at 43 kDa on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Amino acid sequencing revealed that the N terminus of TcVP1 had the highest homology to Trametes versicolor MPG1, lignin peroxidase G (LiPG) IV, Bjerkandera adusta manganese peroxidase 1 (MnP1), and Bjerkandera sp. RBP (12 out of 14 amino acid residues, 86% identity). Mn2+ oxidizing assay revealed that TcVP1 acted like a classical MnP at pH ∼5, while dye-decolorizing and oxidation assays of aromatic compounds revealed that the enzyme acted like a LiP at pH ∼3. TcVP1 showed particularly high decolorizing activity toward azo dyes. Furthermore, coapplication of TcVP1 and the dye-decolorizing peroxidase (DyP) from T. cucumeris Dec 1 was able to completely decolorize a representative anthraquinone dye, Reactive blue 5, in vitro. This decolorization proceeded sequentially; DyP decolorized Reactive blue 5 to light red-brown compounds, and then TcVP1 decolorized these colored intermediates to colorless. Following extended reactions, the absorbance corresponding to the conjugated double bond from phenyl (250–300 nm) decreased, indicating that aromatic rings were also degraded. These findings provide important new insights into microbial decolorizing mechanisms and may facilitate the future development of treatment strategies for dye wastewater.
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A unique dye-decolorizing peroxidase, DyP, from Thanatephorus cucumeris Dec 1: heterologous expression, crystallization and preliminary X-ray analysis.
Acta Crystallographica Section D Biological Crystallography, 2003Co-Authors: Takao Sato, Yasushi Sugano, Shusaku Hara, Takuro Matsui, Sazaki, Shinya Saijo, Tadashi Ganbe, Nobuo Tanaka, Makoto ShodaAbstract:The dye-decolorizing peroxidase DyP is a key enzyme in the decolorizing fungus Thanatephorus cucumeris Dec 1 that degrades azo and antraquinone dyes. The gene dyp from T. cucumeris Dec 1, which has low homology to other peroxidase genes, was cloned and transformed into Aspergillus oryzae and glycosylated DyP was expressed at high levels. Purified DyP was deglycosylated using GST Endo F1 and then crystallized in a strong magnetic field (10 T) at 283 K using ammonium sulfate as precipitant. X-ray diffraction data to 2.96 A resolution collected from a native crystal at the Photon Factory (Tsukuba, Japan) showed that the crystal belonged to the hexagonal space group P6(5)22, with unit-cell parameters a = b = 136.15, c = 363.46 A. The asymmetric unit of the crystal contained four DyP molecules, with a corresponding Matthews coefficient (V(M)) of 2.50 A(3) Da(-1) and a solvent content of 51%. Heavy-atom derivatives of DyP have been obtained and electron-density maps have been calculated. The haem is visible and continuous electron density between the haem and protein clearly indicates the location of the proximal histidine ligand.
Makoto Shoda - One of the best experts on this subject based on the ideXlab platform.
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Purification and characterization of two DyP isozymes from Thanatephorus cucumeris Dec 1 specifically expressed in an air-membrane surface bioreactor.
Journal of bioscience and bioengineering, 2009Co-Authors: Takuya Shimokawa, Makoto Shoda, Yasushi SuganoAbstract:DyP isozymes (DyP2 and DyP3) from the culture fluid of the fungus Thanatephorus cucumeris Dec 1 by air-membrane surface bioreactor were purified and characterized. The characteristics of DyP2 were almost the same as those of a recombinant DyP reported previously, but different from DyP3.
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Efficient dye decolorization and production of dye decolorizing enzymes by the basidiomycete Thanatephorus cucumeris Dec 1 in a liquid and solid hybrid culture.
Journal of bioscience and bioengineering, 2008Co-Authors: Takuya Shimokawa, Makoto Shoda, Mitsuyo Hirai, Yasushi SuganoAbstract:Thanatephorus cucumeris Dec 1, a basidiomycete, is a promising decomposer of several xenobiotics. Air-membrane surface bioreactor culture (AMS culture), a hybrid between solid-state and submerged culture, was used for Dec 1 growth. The optimum temperature for AMS culture (25°C) was lower than that for submerged culture (30°C). Fungal growth was almost the same in AMS culture as in submerged culture at optimum temperatures. However, protein secretion, dye decolorizing peroxidase (DyP) activity, and aryl alcohol oxidase activity, were 18-, 233-, and 36.5-fold higher in AMS culture than in submerged culture, respectively, indicating that AMS culture of Dec 1 was superior to submerged culture. In in vivo dye decolorization tests, 13 of 16 dyes were decolorized by more than 90% within 10 days. Interestingly, under AMS culture, a biofilm was formed; biofilm formation was not, however, essential for DyP and manganese dependent peroxidase (MnP) activities. Although the correlation between DyP activity and water activity was unclear, MnP activity seemed to increase in activity with decreasing water activity, even when no biofilm formation was observed.
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Complete decolorization of the anthraquinone dye Reactive blue 5 by the concerted action of two peroxidases from Thanatephorus cucumeris Dec 1
Applied microbiology and biotechnology, 2006Co-Authors: Yasushi Sugano, Yuko Matsushima, Makoto ShodaAbstract:It is useful to identify and examine organisms that may prove useful for the treatment of dye-contaminated wastewater. Here, we report the purification and characterization of a new versatile peroxidase (VP) from the decolorizing microbe, Thanatephorus cucumeris Dec 1 (TcVP1). The purified TcVP1 after Mono P column chromatography showed a single band at 43 kDa on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Amino acid sequencing revealed that the N terminus of TcVP1 had the highest homology to Trametes versicolor MPG1, lignin peroxidase G (LiPG) IV, Bjerkandera adusta manganese peroxidase 1 (MnP1), and Bjerkandera sp. RBP (12 out of 14 amino acid residues, 86% identity). Mn2+ oxidizing assay revealed that TcVP1 acted like a classical MnP at pH ∼5, while dye-decolorizing and oxidation assays of aromatic compounds revealed that the enzyme acted like a LiP at pH ∼3. TcVP1 showed particularly high decolorizing activity toward azo dyes. Furthermore, coapplication of TcVP1 and the dye-decolorizing peroxidase (DyP) from T. cucumeris Dec 1 was able to completely decolorize a representative anthraquinone dye, Reactive blue 5, in vitro. This decolorization proceeded sequentially; DyP decolorized Reactive blue 5 to light red-brown compounds, and then TcVP1 decolorized these colored intermediates to colorless. Following extended reactions, the absorbance corresponding to the conjugated double bond from phenyl (250–300 nm) decreased, indicating that aromatic rings were also degraded. These findings provide important new insights into microbial decolorizing mechanisms and may facilitate the future development of treatment strategies for dye wastewater.
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A unique dye-decolorizing peroxidase, DyP, from Thanatephorus cucumeris Dec 1: heterologous expression, crystallization and preliminary X-ray analysis.
Acta Crystallographica Section D Biological Crystallography, 2003Co-Authors: Takao Sato, Yasushi Sugano, Shusaku Hara, Takuro Matsui, Sazaki, Shinya Saijo, Tadashi Ganbe, Nobuo Tanaka, Makoto ShodaAbstract:The dye-decolorizing peroxidase DyP is a key enzyme in the decolorizing fungus Thanatephorus cucumeris Dec 1 that degrades azo and antraquinone dyes. The gene dyp from T. cucumeris Dec 1, which has low homology to other peroxidase genes, was cloned and transformed into Aspergillus oryzae and glycosylated DyP was expressed at high levels. Purified DyP was deglycosylated using GST Endo F1 and then crystallized in a strong magnetic field (10 T) at 283 K using ammonium sulfate as precipitant. X-ray diffraction data to 2.96 A resolution collected from a native crystal at the Photon Factory (Tsukuba, Japan) showed that the crystal belonged to the hexagonal space group P6(5)22, with unit-cell parameters a = b = 136.15, c = 363.46 A. The asymmetric unit of the crystal contained four DyP molecules, with a corresponding Matthews coefficient (V(M)) of 2.50 A(3) Da(-1) and a solvent content of 51%. Heavy-atom derivatives of DyP have been obtained and electron-density maps have been calculated. The haem is visible and continuous electron density between the haem and protein clearly indicates the location of the proximal histidine ligand.
Mitsuro Hyakumachi - One of the best experts on this subject based on the ideXlab platform.
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Mechanism of the generation of new somatic compatibility groups within Thanatephorus cucumeris (Rhizoctonia solani).
Microbes and environments, 2013Co-Authors: M.g.b. Saldajeno, Mitsuro HyakumachiAbstract:Single-basidiospore isolates (SBIs) were obtained from field isolates of Thanatephorus cucumeris (Rhizoctonia solani) AG-1 IC and AG-2-2 IV. Formation of distinctive tufts, a recognized feature of heterokaryon synthesis, was observed, and isolates derived from hyphal-tipped tuft hyphae were obtained following pairings between various strains. Three distinctive types of tufts were formed: the fibrous type of mating-compatible homokaryon-homokaryon (Hom-Hom) pairings, the sparse type between heterokaryon-homokaryon (Het-Hom) pairings originating from one parent, and the compact type between Het-Hom pairings originating from different parents. Amplified Fragment Length Polymorphism (AFLP) profile of fingerprints of these tuft isolates verified that they were all heterokaryotic. Because of heterokaryotic vigor, the growth and pathogenicity of the majority of tuft isolates increased compared with their contributing SBIs. New somatic compatibility groups (SCGs) that were different from parental field isolates occurred following heterokaryon formation within T. cucumeris. Tuft isolates produced by Hom-Hom and Het-Hom pairings among isolates of different parents yielded no somatic compatibility with the original parent isolates and a high frequency of new SCGs (62–100%). This was in contrast to those produced by Hom-Hom and Het-Hom pairings among isolates with a common parent that yielded only 12–37% new SCGs. The SCG diversity of R. solani in the field may be attributed to new fitter heterokaryons formed between a heterokaryon of one pair of parents and a homokaryon of another parent pair. This mechanism greatly contributes to genetic diversity in the field and accounts for the failure to recover the expected distribution of SCGs from a field population.
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Mechanism of the Generation of New Somatic Compatibility Groups within Thanatephorus cucumeris (Rhizoctonia solani)
2012Co-Authors: Mary Grace, B. Saldajeno, Mitsuro HyakumachiAbstract:Single-basidiospore isolates (SBIs) were obtained from field isolates of Thanatephorus cucumeris (Rhizoctonia solani) AG-1 IC and AG-2-2 IV. Formation of distinctive tufts, a recognized feature of heterokaryon synthesis, was observed, and isolates derived from hyphal-tipped tuft hyphae were obtained following pairings between various strains. Three distinctive types of tufts were formed: the fibrous type of mating-compatible homokaryon-homokaryon (Hom-Hom) pairings, the sparse type between heterokaryon-homokaryon (Het-Hom) pairings originating from one parent, and the compact type between Het-Hom pairings originating from different parents. Amplified Fragment Length Polymorphism (AFLP) profile of fingerprints of these tuft isolates verified that they were all heterokaryotic. Because of heterokaryotic vigor, the growth and pathogenicity of the majority of tuft isolates increased compared with their contributing SBIs. New somatic compatibility groups (SCGs) that were different from parental field isolates occurred following heterokaryon formation within T. cucumeris. Tuft isolates produced by Hom-Hom and Het-Hom pairings among isolates of different parents yielded no somatic compatibility with the original parent isolates and a high frequency of new SCGs (62–100%). This was in contrast to those produced by Hom-Hom and Het-Hom pairings among isolates with a common parent that yielded only 12–37 % new SCGs. The SCG diversity of R. solani in the field may be attributed to new fitter heterokaryons formed between a heterokaryon of one pair of parents and a homokaryon of another parent pair. This mechanism greatly contributes to genetic diversity in the field and account
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Heterokaryon formation in Thanatephorus cucumeris (Rhizoctonia solani) AG-1 IC.
Mycological Research, 2008Co-Authors: Koji Yamashita, Achmadi Priyatmojo, Takeshi Toda, Mayumi Kubota, Mitsuro HyakumachiAbstract:Approximately 50 single-basidiospore isolates (SBIs) obtained from each of 16 field isolates of Thanatephorus cucumeris AG-1 IC were examined for heterokaryon formation. All SBIs obtained from each field isolate were divided into two mating groups (SBIs-M1 and SBIs-M2), and tufts of mycelia were formed in the contact zone between colonies of paired SBIs-M1 and -M2 based on 0.5 % charcoal agar medium. Tufts were produced from all possible pairing between SBIs from non-parental field isolates. Hyphal anastomosis reactions indicated no cell death and random cell death at the contact cell, and was not related to tuft formation. AFLP phenotypes of SBIs from each field isolate were not identical to each other and were different from their parental field isolate. AFLP phenotypes of the tuft isolates formed from SBIs-M1 and SBIs-M2 from each field isolate were heterokaryotic. Moreover, several SBIs also formed tufts with their parental and non-parental field isolates. AFLP phenotypes of these tuft isolates suggested that they were all heterokaryotic. Results of these experiments suggest that T. cucumeris AG-1 IC is heterothallic and bipolar, and that genetic exchange can occur between homokaryotic and heterokaryotic isolates (Buller phenomenon).
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heterokaryon formation in Thanatephorus cucumeris anastomosis group 2 2 iv
Mycologia, 2006Co-Authors: Takeshi Toda, Mitsuro HyakumachiAbstract:Thirty single basidiospore isolates (SBIs) obtsained from four field isolates of the basidiomycete fungus Thanatephorus cucumeris AG 2-2 IV were examined for heterokaryon formation. SBIs of three of four field isolates (Rh509, 92155 and R94) did not produce a tuft of mycelium in the hyphal interaction zone between paired isolates on 2% charcoal agar. Field isolates Rh509, 92155 and R94 indicated no death of interacting mycelium with their progenies on glass slide and microscopic examination. AFLP (amplified fragment length polymorphism) phenotypes of parent and their SBIs were identical. Field isolates Rh509, 92155 and R94 and their SBIs were homothallic. SBIs obtained from field isolate SA-1 were grouped into two mating types (SBI-M1 and SBI-M2), and a tuft of mycelium was formed between paired SBIs-M1 and -M2. SBIs of field isolate SA-1 indicated that no death and death of interacting mycelium were randomly observed. AFLP phenotypes among SBIs of isolate SA-1 were not identical and were also different f...
J S Nicholson - One of the best experts on this subject based on the ideXlab platform.
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Evaluation of Tobacco Germplasm for Seedling Resistance to Stem Rot and Target Spot Caused by Thanatephorus cucumeris.
Plant Disease, 2008Co-Authors: P. E. Elliott, Ramsey S. Lewis, W. A. Gutierrez, H D Shew, J S NicholsonAbstract:ABSTRACT Stem rot and target spot of tobacco, caused by Rhizoctonia solani and its teleomorph Thanatephorus cucumeris, respectively, can cause serious problems in production of tobacco (Nicotiana tabacum) seedlings. Previous screens for genetic resistance in tobacco have been limited. The objective of this study was to evaluate 97 genotypes composing several classes of tobacco and related Nicotiana spp. for seedling resistance to stem rot and target spot. Significant differences in disease incidence initially were observed among the genotypes for both stem rot and target spot; however, resistance to target spot was not observed when disease pressure was high. Partial resistance to stem rot was observed in several genotypes in repeated tests. These accessions may be useful as a source of resistance to R. solani in future breeding efforts.
Jack Call - One of the best experts on this subject based on the ideXlab platform.
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Characterization and Pathogenicity of Thanatephorus cucumeris from Sugar Beet in Minnesota.
Plant disease, 1997Co-Authors: Carol E. Windels, Rita A. Kuznia, Jack CallAbstract:ABSTRACT In 1993, hymenia of Thanatephorus cucumeris occurred on petioles of sugar beet leaves, but disease was not observed on leaves, crowns, or roots. Of 33 cultures isolated from sugar beet, 28 were identified as Rhizoctonia solani AG-3 (from four fields planted to potatoes in 1992) and five isolates were AG-5 (from one field planted to wheat in 1992). These isolates of R. solani AG-3 and AG-5 were nonpathogenic to moderately pathogenic on sugar beet seedlings (stands ranged from 49 to 95%). The same isolates were nonpathogenic when inoculated on 8-week-old sugar beet roots (root rot indices were ≤1 [0 to 7 scale]). All isolates of R. solani AG-3 (but none of AG-5) formed sclerotia on roots. Disease indices (0 to 4 scale) on potato sprouts at 10°C were low, did not differ significantly (P = 0.05) among isolates and the control in either of two experiments, and averaged 0.9 for 14 isolates of AG-3, 0.5 for three isolates of AG-5, and 0.5 in the control. All isolates of AG-3 (but none of AG-5) formed sc...