Purification and Properties of Benzyl Alcohol Oxidase from Botrytis cinerea

A benzyl alcohol oxidase (BAO) was purified to homogeneity from Botrytis cinerea. The enzyme was found to have a molecular mass of 214 kD with a trimeric structure, and optimal pH and temperature of 5.0 and 30°C, respectively. The enzyme activity was not sensitive to metal ions or to metal ion chelators, while thiol blocking reagents strongly inhibited BAO activity. Sulfur dioxide irreversibly inhibited the enzyme activity and the inhibitory effect of ethanol was weak and reversible. Benzyl alcohol was the most effective alcohol substrate for BAO. Para or meta monosubstituted benzyl alcohol with methyl or methoxy groups were good substrates. BAO also oxidized cinnamyl alcohol, furfuryl alcohol, and some terpenic alcohols· with an alkenyl group near the reactive carbinol. Secondary alcohol, methanol and phenol were not substrates. Product inhibition studies suggested that benzaldehyde and benzyl alcohol were bound at different places to the active site. O₂ was the only electron acceptor identified and Botrytis cinerea benzyl alcohol oxidase was classified .as EC 1.1.3.7 according to stoichiometrical studies. We discuss the metabolic role of BAO in the Botrytis cinerea-grape host-parasite relationship.     

Molecular identification and determination of the trichothecene chemotypes of fungi causing crown and root in different growth stages of wheat in north of Iran

In order to determine the crown and root agents and their mycotoxins produced in different growth stages of wheat including seedling, tillering and heading, sampling was done in north of Iran, during 2011–2012. From 160 isolates of Fusarium, eight species were obtained including F. graminearum, F. culmorum, F. equiseti, F. nygamai, F. semitectum, F. solani, F. acuminatum and F. oxysporum. Sampling at different growth stages showed that F. graminearum was the predominant causal agent of crown and root at the heading stage, whereas other species of Fusarium were mostly observed at the seedling and tillering stages. Moreover, identification of pathogenic species was confirmed using species-specific primers pairs. In F. graminearum isolates, presence of Tri13 gene, responsible for nivalenol (NIV) and deoxynivalenol (DON) mycotoxins biosynthesis, was detected using specific PCR primers. Finally, the ability of trichothecene production of five F. graminearum isolates was confirmed with high-performance liquid chromatography.     

Taxonomy of some common Amphidinium species

The taxonomic status of several well known species belonging to the genus Amphidinium (Dinophyceae) has been re-examined. Studies in culture and observations on the micro-anatomy of these organisms suggest that the number of species can be reduced to two, A. klebsii and A. carterae. They may be distinguished on the basis of fundamental differences in chloroplast morphology.     

Asymmetric reduction of aryl ketones with a new isolate Rhodotorula sp. AS2.2241

A yeast strain, Rhodotorula sp. AS2.2241, capable of reducing acetophenone and α-bromoacetophenone with high stereoselectivity, was isolated from soil samples through a novel screening procedure in which acetophenone was supplied in vapor state as the sole carbon and energy source. The biosynthesis of the ketone reductase in the yeast cells reached a maximum of 41.0 U/l at 20 h of cultivation. The reductase isolated from the Rhodotorula sp. cells was partially purified by 52.6-fold through a single column chromatography of DEAE–cellulose. The catalytic performance of the partially purified reductase was examined, and the highest activity was observed at pH 6.5 and 50 °C. The short-chain alkyl aldehydes such as acetaldehyde and those aldehydes or ketones with a benzoyl group were found to be good substrates for the reductase. In the preparative bioreductions of 50 mM acetophenone and 2 mM α-bromoacetophenone using resting cells of Rhodotorula sp. AS2.2241, (S)-(−)-1-phenylethanol (>99.5% enantiomeric excess (e.e.), 34.7% yield) and (R)-(−)-2-bromo-1-phenylethanol (>99.9% e.e., 19.9% yield) were obtained, respectively.     

A Novel Electrochemically Active and Fe(III)-reducing Bacterium Phylogenetically Related to Clostridium butyricum Isolated from a Microbial Fuel Cell

An obligatory anaerobic bacterium was isolated from a mediator-less microbial fuel cell using starch processing wastewater as the fuel and designated as EG3. The isolate was Gram-positive, motile and rod (2.8–3.0 μm long, 0.5–0.6 μm wide). The partial 16S rRNA gene sequence and analysis of the cellular fatty acids profile suggested that EG3 clusters with Clostridium sub-phylum and exhibited the highest similarity (98%) with Clostridium butyricum. The temperature and pH optimum for growth were 37°C and 7.0, respectively. The major products of glucose and glucose/Fe(O)OH metabolism were lactate, formate, butyrate, acetate, CO₂and H₂. Growth was faster at the initial phase and the cell yield was higher when the medium was supplemented with Fe(O)OH than without Fe(O)OH. These results suggest that Fe(III) ion is utilised as an electron sink. Cyclic voltammetry showed that Clostridium butyricum EG3 cells were electrochemically active. It is a novel characteristic of strict anaerobic Gram-positive bacteria.     

Onychiurid species from Wanda Mountains in China,with descriptions of two new species (Collembola,Onychiuridae)

A checklist of onychiurid species from the Wanda Mountains in China is presented. Eighteen species belonging to twelve genera have been found, including two new species. Bionychiurus qinglongensis sp. n. can be easily distinguished from other known species of the genus by the absence of pseudocelli on Th. I tergum and fewer number of vesicles in postantennal organ. Onychiurus heilongjiangensis sp. n. is diagnosed by pseudocellar formulae as 32/133/33352 dorsally and 3/011/31120 ventrally, parapseudocellar formula as 0/000/111001+1^m, ratio of anal spine/unguis as 0.6, unguiculus without inner basal lamella, and male ventral organ absent.     

Biomineralization of 2,3,4,6-Tetrachlorophenol by Bacillus sp. and Staphylococcus sp. Isolated from Secondary Sludge of Pulp and Paper Mill

Three 2,3,4,6-tetrachlorophenol (2,3,4,6-TeCP)-mineralizing bacteria were isolated from the secondary sludge of a pulp and paper industry. The isolates used 2,3,4,6-TeCP as a source of carbon and energy and were capable of degrading this compound, as indicated by stoichiometric release of chloride and biomass formation. Based on 16S rRNA gene sequence analysis, the bacteria were identified as Bacillus megaterium (CL3), Staphylococcus suciri (CL10), and Bacillus thuringensis (CL11). High-performance liquid chromatography (HPLC) analysis revealed that these isolates were able to degrade 2,3,4,6-TeCP at higher concentrations (600 mg/L or 2.5 mM). A consortia of the isolates completely removed 2,3,4,6-TeCP from the sludge obtained from a pulp and paper mill within 2 weeks when supplemented at a rate of 100 mg/L or 0.43 mM. A bacterial consortium also significantly reduced absorbable organic halogen (AOX) and extractable organic halogen (EOX) by 63% and 68%, respectively, from the sludge. These isolates have a high potential to remove 2,3,4,6-TeCP and may be used for remediation of pulp paper mill waste containing 2,3,4,6-TeCP.