The Strain Seismograms of P- and S-Waves of a Local Event Recorded by Four-Gauge Borehole Strainmeter

At a sampling rate of 100 samples per second, the YRY-4 four-gauge borehole strainmeters (FGBS) are capable of recording transient strains caused by seismic waves such as P and S waves or strain seismograms. At such a high sampling rate, data from the YRY-4 strainmeters demonstrate fairly satisfactory self-consistency. The strain tensor seismograms demonstrate the senses of motion of P waves, that is, the type of seismic wave travels in the direction of the maximum normal strain change. The observed strain patterns of S waves significantly differ from those of P waves and should contain information about the source mechanism. Spectrum analysis shows that the strain seismograms are consistent with conventional broadband seismograms from the same site

Regulation of MAL1⁺ gene expression encoding maltase in Schizosaccharomyces pombe by added inositol

143-147In this study, the effects of inositol addition on maltase activity and expression of MAL1⁺ gene encoding maltase in Schizosaccharomyces pombe were investigated. The maximum specific maltase activity was observed, when the concentration of inositol reached 6.0 μg/ml in the synthetic medium containing 2.0% glucose. At 1.0 μg/ml inositol concentration, the maltase activity continuously decreased, as initial glucose concentration was higher than 0.1%. mRNA encoding maltase and phosphatidylinositol (PI) content were higher in the cells grown in the synthetic medium with 6.0 μg/ml of inositol and 2.0% glucose than those with 1.0 μg/ml of inositol. These results demonstrated that higher inositol concentration in the synthetic medium could derepress MAL1⁺ gene expression in S. pombe and PI might be involved in derepression of MAL1⁺ gene expression in S. pombe probably by PI-type signalling pathway

Expression of the inulinase gene from the marine-derived Pichia guilliermondii in Saccharomyces sp. W0 and ethanol production from inulin

International audienceIt has been confirmed that Saccharomyces sp. W0 can produce high concentration of ethanol. In this study, the INU1 gene cloned from the marine-derived Pichia guilliermondii was transformed into uracil mutant of Saccharomyces sp. W0. The positive transformant Inu-66 obtained could produce 34.2 U ml-1 of extracellular inulinase within 72 h of cultivation. It was found that 15.2 U of inulinase activity per one gram of inulin was suitable for inulin hydrolysis and ethanol production by the transformant Inu-66. During the small-scale fermentation, 13.7 ml of ethanol in 100 ml of medium was produced and 99.1% of the added inulin was utilized by the transformant. During the 2 l fermentation, 14.9% (v/v) of ethanol was produced from inulin and 99.5% of the added inulin was converted into ethanol, CO2 and cell mass

Added inositol regulates invertase secretion and glucose-repressed SUC2 gene expression in Saccharomyces sp. W4

152-156The effect of inositol supplementation on glucose derepression, invertase secretion and SUC2 gene expression in Saccharomyces sp. W4 was studied. Invertase secretion was repressed, when the yeast cells, grown the synthetic medium without inositol (I⁻ medium) contained more than 0.2% (w/v) initial concentration of glucose. However, in the same medium plus inositol (I⁺ medium, inositol conc. 100 μg/100 ml), invertase secretion was repressed only at glucose concentrations higher than 2.0% (w/v). Results showed that secreted invertase activity increased only in the I⁺ medium, whereas intracellular invertase activity remained constant in both media during the cell, growth. The mRNA encoding secreted invertase was higher in the glucose-derepressed cells grown in the I⁺ medium than in the glucose-repressed cells grown in the I⁻ medium. Similarly, phosphatidylinositol (PI) content was significantly higher in the cells grown in the I⁺ medium than in the I⁻ medium. These results indicated that PI might be involved in the glucose derepression, invertase secretion and SUC2 gene expression at the transcriptional level in the yeast

<span style="mso-bidi-language:HI">Purification and characterization of trehalose-6-phosphate synthase from <i><span style="mso-bidi-language:HI">Saccharomycopsis </span></i><i><span style="mso-bidi-font-family:Arial;mso-bidi-language:HI">fibuligera </span></i><span style="mso-bidi-language:HI">A<span style="mso-bidi-font-family:Arial; mso-bidi-language:HI">11 </span></span></span>

289-294Mutant <span style="mso-bidi-font-family: Arial;mso-bidi-language:HI">A11, a mutant of Saccharomycopsis fibuligera Sdu with low acid and neutral trehalase was found to <span style="mso-bidi-font-family:Arial; mso-bidi-language:HI">accumulate over 18% (w/w) trehalose <span style="mso-bidi-font-family:Arial;mso-bidi-language: HI">from starch in its cells. In this study, trehalose-6-phosphate synthase <span style="mso-bidi-font-family: Arial;mso-bidi-language:HI">(Tps1) was purified to homogeneity from this mutant, with a 30-fold increase in the specific enzyme activity, as compared to the concentrated cell-free extract, from initial cells. The molecular mass of the purified enzyme as determined by SDS-PAGE was 66 <span style="mso-bidi-font-family:Arial;mso-bidi-language: HI">kDa. The optimum pH and temperature of the purified enzyme were 6.6 and 37°C, respectively. The enzyme was activated by Ca<span style="mso-bidi-font-family: Arial;mso-bidi-language:HI">2<span style="mso-bidi-language: HI">+, K+ and Mg2+, with K+ showing the highest activation at 35 mM. On the other hand, Mn<span style="mso-bidi-font-family:Arial;mso-bidi-language: HI">2+, Cu<span style="mso-bidi-font-family: Arial;mso-bidi-language:HI">2<span style="mso-bidi-language: HI">+, Fe3+, Hg2+ and Co<span style="mso-bidi-font-family:Arial;mso-bidi-language: HI">2+ inhibited the enzyme. The enzyme was also strongly inhibited by protease inhibitors such as iodoacetic acid, EOTA and PMSF. </span

Identification, Characterization, and Expression Analysis of a FMRFamide-Like Peptide Gene in the Common Chinese Cuttlefish (Sepiella japonica)

The peptide FMRFamide is one of the well-known peptides involved in multiple physiological processes in the phylum Mollusca. In this study, a FMRFamide gene (GenBank accession No. KJ933411) was identified in a cuttlefish species called Sepiella japonica and was designated as SjFMRFamide. The total length of the SjFMRFamide sequence was found to be 1880 bp while the open reading frame contained 996 bp encoding a protein of 331 amino acid residues with a predicted isoelectric point (pI) and molecular weight (MW) of 9.18 and 38.8 kDa along with a 333 bp 5′-untranslated region (UTR) and 551 bp 3′-UTR. The deduced SjFMRFamide precursor protein contains one signal peptide and expresses four kinds FMRFamide-related peptides including a single copy of FLRFamide, ALSGDAFLRFamide, and FIRFamide and multiple copies of FMRFamide. Results of phylogenetic relation analysis strongly indicated that the sequence of this gene shares high identity with the genes of known FMRFamides. Spatial expression analysis indicated the highest mRNA expression of SjFMRFamide in the brain of male and female cuttlefishes among the eight tissues analyzed. An in situ hybridization assay of the brain indicated that SjFMRFamide was transcribed in several functional lobes, which suggests that it might be related to many physiological regulatory mechanisms. This is the first study describing FMRFamide in S. japonica and the results may contribute to future studies of neuropeptide evolution or may prove useful for the development of aquaculture methods for this cuttlefish species

Properties of alkaline protease genetically engineered on cell surface of the yeast Yarrowia lipolytica

International audienceALP2 gene encoding alkaline protcase cloned from Aureobasidium pullulans HN2-3 was ligated into the surface display plasmid and expressed in the cells of the yeast Yarrowia lipolytica. The expressed alkaline protease was immobilized on the yeast cells. The activity of the immobilized enzyme with 6 x His tag was found to be significantly higher than that of without 6 x His tag. The immobilized enzyme showed lower optimal temperature and a lower affinity for azocasein than the free enzyme purified from A. pullulans HN2-3. The thermal stability of the immobilized enzyme enhanced and the pH stability decreased, compared to that of the free enzyme

Properties of alkaline protease genetically engineered on cell surface of the yeast Yarrowia lipolytica

International audienceALP2 gene encoding alkaline protcase cloned from Aureobasidium pullulans HN2-3 was ligated into the surface display plasmid and expressed in the cells of the yeast Yarrowia lipolytica. The expressed alkaline protease was immobilized on the yeast cells. The activity of the immobilized enzyme with 6 x His tag was found to be significantly higher than that of without 6 x His tag. The immobilized enzyme showed lower optimal temperature and a lower affinity for azocasein than the free enzyme purified from A. pullulans HN2-3. The thermal stability of the immobilized enzyme enhanced and the pH stability decreased, compared to that of the free enzyme

Properties of alkaline protease genetically engineered on cell surface of the yeast <i style="">Yarrowia lipolytica</i>

294-298ALP2 gene encoding alkaline protease cloned from Aureobasidium pullulans HN2-3 was ligated into the surface display plasmid and expressed in the cells of the yeast Yarrowia lipolytica. The expressed alkaline protease was immobilized on the yeast cells. The activity of the immobilized enzyme with 6 His tag was found to be significantly higher than that of without 6 His tag. The immobilized enzyme showed lower optimal temperature and a lower affinity for azocasein than the free enzyme purified from A. pullulans HN2-3. The thermal stability of the immobilized enzyme enhanced and the pH stability decreased, compared to that of the free enzyme

Genetic modification of the marine-derived yeast Yarrowia lipolytica with high-protein content using a GPI-anchor-fusion expression system

International audienceYarrowia lipolytica SWJ-1b isolated from the marine fish gut was found to contain 47.6 g of crude protein per 100 g of cell dry weight and had potential use as single cell protein. When the gene encoding enhanced green fluorescent protein (EGFP) was inserted into the surface display plasmid pINA1317-YlCWP110 and expressed in uracil mutant Of Y. lipolytica SWJ-Ib, the corresponding protein was successfully displayed on the cell surface, and 100% of the yeast cells exhibited the anchored target proteins. We found that yeast cells displaying EGFP were similar to those of Y. lipolytica SWJ-1b. Furthermore, C-18:1 and C-18:3 fatty acids biosynthesis in the marine yeast cells displaying the heterologous EGFP was weakly enhanced compared with that in its wild-type. The results suggest that the marine-derived Y. lipolytica SWJ-1b can be armed with the heterologous protein by the genetic modification and further used as single cell protein