RESUMO
Ten strains which were characterized by the formation of ballistoconidia, the absence of xylose in whole-cell hydrolysates, the presence of Q-9 as the major ubiquinone isoprenologue, the inability to ferment sugars and positive diazonium blue B and urease reactions were isolated from plant samples collected in Thailand. These isolates were closely related to Bensingtonia phyllada based on the analysis of 18S rDNA sequences. On the basis of the morphological, physiological and chemotaxonomic properties, the 10 isolates were assigned to the genus Bensingtonia. DNA complementarity showed that these isolates were genetically distinct from known species of the genus Bensingtonia. The isolates are described as Bensingtonia thailandica sp. nov. The type strain is strain TY-138T (= JCM 10651T).
Assuntos
Basidiomycota/classificação , Filogenia , Folhas de Planta/microbiologia , Basidiomycota/genética , Basidiomycota/isolamento & purificação , Metabolismo dos Carboidratos , DNA Fúngico/genética , DNA Ribossômico/genética , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Tailândia , Clima TropicalRESUMO
The gene for carboxymethyl cellulose-degrading enzyme (endoglucanase) from Clostridium josui (FERM P-9684) was cloned in Escherichia coli HB101 with pBR322. A 5.6-kilobase-pair HindIII fragment encoding an endoglucanase was hybridized with C. josui chromosomal DNA. The size of the cloned DNA fragment was reduced with PvuII, and the resulting active fragment (2 kilobase pairs, with restriction sites of EcoRI and PstI) was ligated into pUC118 at the SmaI sites (pUCJ1). The endoglucanase production by E. coli JM103(pUCJ1) in Luria-Bertani broth was enhanced up to approximately three times by maintaining the pH at 6.5 and using 80 mM NaCl.
Assuntos
Proteínas de Bactérias/genética , Celulase/genética , Clostridium/genética , Escherichia coli/genética , Genes Bacterianos , Proteínas de Bactérias/biossíntese , Celulase/biossíntese , Celulase/metabolismo , Clonagem Molecular , Clostridium/enzimologia , Clostridium/crescimento & desenvolvimento , DNA Bacteriano/isolamento & purificação , Plasmídeos , Mapeamento por Restrição , Transformação BacterianaRESUMO
An enzyme active against carboxymethyl cellulose (CMC) was purified from the stationary-phase-culture supernatant of Clostridium josui grown in a medium containing ball-milled cellulose. The purification in the presence of 6 M urea yielded homogeneous enzyme after an approximately 50-fold increase in specific activity and a 13% yield. The enzyme had a molecular mass of 45 kilodaltons. The optimal temperature and pH of the enzyme against CMC were 60 degrees C and 6.8, respectively. The enzyme hydrolyzed cellotetraose, cellopentaose, and cellohexaose to cellobiose and cellotriose but did not hydrolyze cellobiose or cellotriose. A microcrystalline cellulose, Avicel, was also hydrolyzed significantly, but the extent of hydrolysis was remarkably less than that of CMC. On the basis of these results, the enzyme purified here is one of the endo-1,4-beta-glucanases. The N-terminal amino acid sequence of the enzyme is Tyr-Asp-Ala-Ser-Leu-Lys-Pro-Asn-Leu-Gln-Ile-Pro-Gln-Lys-Asn-Ile-Pro-Asn- Asn-Asp-Ala-Val-Asn-Ile-Lys.
