Yeast cell mortality related to a high-pressure shift: occurrence of cell membrane permeabilization

The shrinkage of yeast cells caused by high-pressure treatment (250 MPa, 15 min) was investigated using direct microscopic observation. A viable staining method after treatment allowed the volume variation of two populations to be distinguished: an irreversible volume decrease (about 35% of the initial volume) of pressure-inactivated cells during pressure holding time, and viable cells, which were less affected. A mass transfer was then induced during high-pressure treatment. Causes of this transfer seem to be related to a pressure-induced membrane permeabilization, allowing a subsequent leakage of internal solutes, where three ions (Na+, K+ and Ca2+), plus endogenous glycerol, were verified. This glycerol leakage was found to occur after yeast pressurization in a medium having low water activity, although the yeast was not inactivated. All these observations lead to the hypothesis that pressure-induced cell permeabilization could be the cause of yeast inactivation under pressure.     

Schriftenschau

Ohne Zusammenfassung     

Modeling Tetrahymena thermophila growth and protease production

Oxygen concentrations stimulated growth (maximum number of cells) and protease secretion by Tetrahymena thermophila. Agitation and aeration conditions for growth and protease secretion were optimised by a central composite design. The best optimised combination was a stirrer speed of 338 rpm and an aeration of 1 vvm. Journal of Industrial Microbiology & Biotechnology (2000) 25, 58–61. Received 24 September 1999/ Accepted in revised form 06 March 2000     

Identification of a rice RNA- and microtubule-binding protein as the multifunctional protein, a peroxisomal enzyme involved in the beta -oxidation of fatty acids.

The control of subcellular mRNA localization and translation is often mediated by protein factors that are directly or indirectly associated with the cytoskeleton. We report the identification and characterization of a rice seed protein that possesses both RNA and microtubule binding activities. In vitro UV cross-linking assays indicated that this protein binds to all mRNA sequences tested, although there was evidence for preferential binding to RNAs that contained A-C nucleotide sequence motifs. The protein was purified to homogeneity using a two-step procedure, and amino acid sequencing identified it as the multifunctional protein (MFP), a peroxisomal enzyme known to possess a number of activities involved in the beta-oxidation of fatty acids. The recombinant version of this rice MFP binds to RNA in UV cross-linking and gel mobility shift experiments, co-sediments specifically with microtubules, and possesses at least two enzymatic activities involved in peroxisomal fatty acid beta-oxidation. Taken together these data suggest that MFP has an important role in mRNA physiology in the cytoplasm, perhaps in regulating the localization or translation of mRNAs through an interaction with microtubules, in addition to its peroxisomal function.     

Amperometric alcohol electrode with extended linearity and reduced interferences.

Conventional amperometric alcohol electrodes were constructed with oxygen- and hydrogen peroxide-base sensors and a much improved electrode was designed by placing a hydrophobic, gas-permeable membrane over the conventional hydrogen peroxide-based alcohol electrode. The immobilization of alcohol oxidase with glutaraldehyde was also studied and optimized. The upper linear ranges of the conventional and newly designed alcohol electrodes were 0.02 and 0.5% ethanol, respectively. The hydrophobic membrane of the new design eliminated the classical electrochemical interferences of hydrogen peroxide-based electrodes and the typical pH dependence of enzymatic systems.     

Procedure and Analysis of a Useful Method in Determining Mycelial Dry Weights from Agar Plates

The evaluation of growth by dry weight determination of fungus mycelium for agar plates was examined. The data obtained were statistically analyzed. This method was shown to be sufficiently accurate to be used as an investigative tool.