{Reference Type}: Journal Article
{Author}: Kontkanen, H.; Westerholm-Parvinen, A.; Saloheimo, M.; Bailey, M.; Ratto, M.; Mattila, I.; Mohsina, M.; Kalkkinen, N.; Nakari-Setala, T.; Buchert, J.
{Year}: 2009
{Title}: Novel Coprinopsis cinerea polyesterase that hydrolyzes cutin and suberin
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=19201950&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Environ Microbiol
{Volume}: 75
{Issue}: 7
{Pages}: 2148-57
{DOI}: 10.1128/AEM.02103-08
{Date Displayed}: 2009 Apr
{Date}: 2009-04-01
{Type of Work}: Journal Article; Research Support, Non-U.S. Gov't
{Accession Number}: 19201950
{Keywords}: Agaricales/*enzymology/*genetics; Butyrates/metabolism; Chromatography, Affinity; Cloning, Molecular; DNA, Fungal/chemistry/genetics; Enzyme Stability; Gene Expression; Hydrogen-Ion Concentration; Hydrolases/chemistry/*genetics/isolation & purification/*metabolism; Kinetics; *Lipids; Membrane Lipids/*metabolism; Molecular Sequence Data; Molecular Weight; Sequence Analysis, DNA; Substrate Specificity; Temperature; Trichoderma/genetics
{Abstract}: Three cutinase gene-like genes from the basidiomycete Coprinopsis cinerea (Coprinus cinereus) found with a similarity search were cloned and expressed in Trichoderma reesei under the control of an inducible cbh1 promoter. The selected   transformants of all three polyesterase constructs showed activity with p-nitrophenylbutyrate, used as a model substrate. The most promising transformant of the cutinase CC1G_09668.1 gene construct was cultivated in a laboratory fermentor, with a production yield of 1.4 g liter(-l) purified protein. The expressed cutinase (CcCUT1) was purified to homogeneity by immobilized metal affinity chromatography exploiting a C-terminal His tag. The N terminus of the enzyme was found to be blocked. The molecular mass of the purified enzyme was determined to be around 18.8 kDa by mass spectrometry. CcCUT1 had higher activity on shorter (C(2) to C(10)) fatty acid esters of p-nitrophenol than on longer ones, and it also exhibited lipase activity. CcCUT1 had optimal activity between   pH 7 and 8 but retained activity over a wide pH range. The enzyme retained 80% of its activity after 20 h of incubation at 50 degrees C, but residual activity decreased sharply at 60 degrees C. Microscopic analyses and determination of released hydrolysis products showed that the enzyme was able to depolymerize apple cutin and birch outer bark suberin.
{Author Address}: VTT, P.O. Box 1000, FI-02044 Espoo, Finland.
{Language}: eng

{Reference Type}: Journal Article
{Author}: Warmink, J. A.; van Elsas, J. D.
{Year}: 2009
{Title}: Migratory response of soil bacteria to Lyophyllum sp. strain Karsten in soil microcosms
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=19286795&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Environ Microbiol
{Volume}: 75
{Issue}: 9
{Pages}: 2820-30
{DOI}: 10.1128/AEM.02110-08
{Date Displayed}: 2009 May
{Date}: 2009-05-01
{Type of Work}: Journal Article
{Accession Number}: 19286795
{Keywords}: *Agaricales; Bacteria/classification/genetics/*growth & development/isolation & purification; *Bacterial Physiological Phenomena; DNA Fingerprinting; DNA, Bacterial/chemistry/genetics; DNA, Ribosomal/chemistry/genetics; Genes, rRNA; *Locomotion; Molecular Sequence Data; Phylogeny; RNA, Bacterial/genetics; RNA, Ribosomal, 16S/genetics; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; *Soil Microbiology
{Abstract}: In this study, the selection of bacteria on the basis of their migration via fungal hyphae in soil was investigated in microcosm experiments containing Lyophyllum sp. strain Karsten (DSM2979). One week following inoculation with a bacterial community obtained from soil, selection of a few specific bacterial types was noticed at 30 mm in the growth direction of Lyophyllum sp. strain Karsten in sterile soil. Cultivation-based analyses showed that the migration-proficient types encompassed 10 bacterial groups, as evidenced by (GTG)(5) genomic fingerprinting as well as 16S rRNA gene sequencing. These were (>97% similarity) Burkholderia terrae BS001, Burkholderia sordidicola BS026, Burkholderia sediminicola BS010, and Burkholderia phenazinium BS028; Dyella japonica BS013, BS018, and BS021; "Sphingoterrabacterium pocheensis" BS024; Sphingobacterium daejeonense BS025; and Ralstonia basilensis BS017. Migration as   single species was subsequently found for B. terrae BS001, D. japonica BS018 and   BS021, and R. basilensis BS017. Typically, migration occurred only when these organisms were introduced at the fungal growth front and only in the direction of hyphal growth. Migration proficiency showed a one-sided correlation with the presence of the hrcR gene, used as a marker for the type III secretion system (TTSS), as all single-strain migrators were equipped with this system and most non-single-strain migrators were not. The presence of the TTSS stood in contrast   to the low prevalence of TTSSs within the bacterial community used as an inoculum (<3%). Microscopic examination of B. terrae BS001 in contact with Lyophyllum sp.   strain Karsten hyphae revealed the development of a biofilm surrounding the hyphae. Migration-proficient bacteria interacting with Lyophyllum sp. strain Karsten may show complex behavior (biofilm formation) at the fungal tip, leading   to their translocation and growth in novel microhabitats in soil.
{Author Address}: Department of Microbial Ecology, Centre for Ecological and Evolutionary Studies,   University of Groningen, Kerklaan 30, 9750RA Haren, The Netherlands.
{Language}: eng

{Reference Type}: Journal Article
{Author}: Tsukihara, T.; Honda, Y.; Sakai, R.; Watanabe, T.; Watanabe, T.
{Year}: 2008
{Title}: Mechanism for oxidation of high-molecular-weight substrates by a fungal versatile peroxidase, MnP2
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=18326680&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Environ Microbiol
{Volume}: 74
{Issue}: 9
{Pages}: 2873-81
{DOI}: 10.1128/AEM.02080-07
{Date Displayed}: 2008 May
{Date}: 2008-05-01
{Type of Work}: Journal Article; Research Support, Non-U.S. Gov't
{Accession Number}: 18326680
{Keywords}: Amino Acid Substitution/genetics; Anthraquinones/*metabolism; Benzyl Alcohols/*metabolism; DNA, Fungal/chemistry/genetics; Enzyme Stability; Fungal Proteins/genetics/metabolism; Hydrogen Peroxide/metabolism; Hydrogen-Ion Concentration; Kinetics; Manganese/metabolism; Models, Molecular; Mutant Proteins/genetics/metabolism; Mutation, Missense; Oxidation-Reduction; Peroxidases/chemistry/genetics/isolation & purification/*metabolism; Pleurotus/genetics/*metabolism; Polymers/*metabolism; Protein Structure, Tertiary; Ribonuclease, Pancreatic/*metabolism; Substrate Specificity
{Abstract}: Unlike general peroxidases, Pleurotus ostreatus MnP2 was reported to have a unique property of direct oxidization of high-molecular-weight compounds, such as Poly R-478 and RNase A. To elucidate the mechanism for oxidation of polymeric substrates by MnP2, a series of mutant enzymes were produced by using a homologous gene expression system, and their reactivities were characterized. A mutant enzyme with an Ala substituting for an exposing Trp (W170A) drastically lost oxidation activity for veratryl alcohol (VA), Poly R-478, and RNase A, whereas the kinetic properties for Mn(2+) and H(2)O(2) were substantially unchanged. These results demonstrated that, in addition to VA, the high-molecular-weight substrates are directly oxidized by MnP2 at W170. Moreover, in the mutants Q266F and V166/168L, amino acid substitution(s) around W170 resulted in a decreased activity only for the high-molecular-weight substrates. These results, along with the three-dimensional modeling of the mutants, suggested that the mutations caused a steric hindrance to access of the polymeric substrates to W170. Another mutant, R263N, contained a newly generated N glycosylation site and showed a higher molecular mass in sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Interestingly, the R263N mutant exhibited an increased reactivity with VA and high-molecular-weight substrates. The existence of an additional carbohydrate modification and the catalytic properties in this mutant are discussed. This is the first study of a direct mechanism for oxidation of high-molecular-weight substrates by a fungal peroxidase using a homologous gene expression system.
{Author Address}: Research Institute for Sustainable Humanosphere, Kyoto University, Gokasho Uji, Kyoto 611-0011, Japan.
{Language}: eng

{Reference Type}: Journal Article
{Author}: Murata, H.; Babasaki, K.; Saegusa, T.; Takemoto, K.; Yamada, A.; Ohta, A.
{Year}: 2008
{Title}: Traceability of Asian Matsutake, specialty mushrooms produced by the ectomycorrhizal basidiomycete Tricholoma matsutake, on the basis of retroelement-based DNA markers
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=18281433&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Environ Microbiol
{Volume}: 74
{Issue}: 7
{Pages}: 2023-31
{DOI}: 10.1128/AEM.02411-07
{Date Displayed}: 2008 Apr
{Date}: 2008-04-01
{Type of Work}: Journal Article; Research Support, Non-U.S. Gov't
{Accession Number}: 18281433
{Keywords}: Agaricales/classification/*genetics/physiology; DNA, Fungal/genetics; Genetic Markers/*genetics; Molecular Sequence Data; Mycorrhizae/*genetics/physiology; Polymerase Chain Reaction; Retroelements/*genetics
{Abstract}: The ectomycorrhizal basidiomycete Tricholoma matsutake produces commercially valuable fruit bodies, matsutake, in forests. Here we report a PCR system targeting retroelement integration sites to differentiate among individual Asian   isolates of T. matsutake based on their geographical origins, such as Japan, the   area of South Korea through North Korea, the northeastern provinces of China, and the area of the southwestern provinces of China through Bhutan. The overall misjudgment rate of the analytical system was approximately 5% based on 95 samples of T. matsutake examined including those from cultures and from commodities. We also provide evidence that T. matsutake isolates grown throughout the Far East, including the northeastern provinces of China, are closely related   to each other while distinct from those in the area of the southwestern provinces of China through Bhutan. The method allows us to trace back geographical origins   of Asian matsutake, thus contributing to food safety, appropriate tariffs, and proper price setting.
{Author Address}: Department of Applied Microbiology and Mushroom Sciences, Forestry and Forest Products Research Institute, Matsunosato 1, Tsukuba 305-8687, Japan. murmur@ffpri.affrc.go.jp
{Language}: eng