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1.
Binding of HMG14 non-histone protein to histones H2A, H2B, H1 and DNA in reconstituted chromatin 总被引:1,自引:0,他引:1
E Espel J Bernués J A Pérez-Pons E Querol 《Biochemical and biophysical research communications》1985,132(3):1031-1037
The interaction between calf thymus HMG14 and rat liver chromatin components has been studied via reconstitution and chemical cross-linking. Selective labeling of HMG14 with photoactivable reversible heterobifunctional reagents has allowed a clear identification of the histones interacting with it (histones H2A, H2B and H1). These results are not dependent on whether the chromatin samples used were bulk chromatin, mononucleosomes, or core particles (for H2A and H2B). In addition to histone proteins, DNA also seems to be involved in HMG14 attachment to nucleosome. 相似文献
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Summary Genes as POT1, HSP104 and SSA3, which are late expressed in laboratory culture conditions are expressed only during the first few days in microvinifications in wine yeast cells. This effect is probably due to the different growth conditions and leads to useless levels of enzyme activity for a reporter gene. However the ACT1 promoter, which is constitutively expressed in laboratory conditions, produces sufficient amounts of enzyme activity in late fermentation phases. 相似文献
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The actinomycete Streptomyces sp. QM-B814 is able to use polymeric glucans as sole carbon source by the induction of specific extracellular enzymatic systems. Carboxymethylcellulose and barley -glucan were the most effective inducers of endoglucanase activities towards -(1,4) and mixed -(1,3)-(1,4) linkages. Avicelase activity was induced by Avicel and carboxymethylcellulose. The higher activity towards filter paper (cellulase) was measured in cultures growing in Avicel, carboxymethylcellulose, and barley -glucan. Laminarinase activity was induced only by laminarin and lichenan failed to induce significant activity levels against any of the substrates tested. 相似文献
4.
Construction of a recombinant wine yeast strain expressing beta-(1,4)-endoglucanase and its use in microvinification processes. 总被引:1,自引:1,他引:0 下载免费PDF全文
A genetic transformation system for an industrial wine yeast strain is presented here. The system is based on the acquisition of cycloheximide resistance and is a direct adaptation of a previously published procedure for brewing yeasts (L. Del Pozo, D. Abarca, M. G. Claros, and A. Jiménez, Curr. Genet. 19:353-358, 1991). Transformants arose at an optimal frequency of 0.5 transformant per microgram of DNA, are stable in the absence of selective pressure, and produce wine in the same way as the untransformed industrial strain. By using this transformation protocol, a filamentous fungal beta-(1,4)-endoglucanase gene has been expressed in an industrial wine yeast under the control of the yeast actin gene promoter. Endoglucanolytic wine yeast secretes the fungal enzyme to the must, producing a wine with an increased fruity aroma. 相似文献
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Sílvia Bronsoms Josep Villanueva Francesc Canals Enrique Querol Francesc X Aviles 《European journal of biochemistry》2003,270(17):3641-3650
Protein folding can be modulated in vivo by many factors. While chaperones act as folding catalysts and show broad substrate specificity, some pro-peptides specifically facilitate the folding of the mature protein to which they are bound. Potato carboxypeptidase inhibitor (PCI), a 39-residue protein carboxypeptidase inhibitor, is synthesized in vivo as a precursor protein that includes a 27-residue N-terminal and a seven-residue C-terminal pro-regions. In this work the disulfide-coupled folding of mature PCI in vitro has been compared with that of the same protein extended with either the N-terminal pro-sequence (ProNtPCI) or both N- and C-terminal pro-sequences (ProPCI), and also with the N-terminal pro-sequence in trans (ProNt + PCI). No significant differences can be observed in the folding kinetics or efficiencies of all these molecules. In addition, in vivo folding studies in Escherichia coli have been performed using wild-type PCI and three PCI mutant forms with and without the N-terminal pro-sequence, the mutations had been previously reported to affect folding of the PCI mature form. The extent to which the 'native-like' form was secreted to the media by each construction was not affected by the presence of the N-terminal pro-sequence. These results indicate that PCI does not depend on the N-terminal pro-sequence for its folding in both, in vitro and in vivo in E. coli. However, structural analysis by spectroscopy, hydrogen exchange and limited proteolysis by mass spectrometry, indicate the capability of such N-terminal pro-sequence to fold within the precursor form. 相似文献
7.
A. Planas M. Juncosa A. Cayetano E. Querol 《Applied microbiology and biotechnology》1992,37(5):583-589
Summary A new purification procedure for endo-\-1,3-1,4-d-glucanase from Bacillus licheniformis is described. The secreted enzyme was purified both from B. licheniformis and from recombinant Escherichia coli harbouring the cloned gene by ion exchange chromatography on a CM-Sepharose matrix at pH 5.6. The mature enzyme was resistant to proteolysis by trypsin and chymotrypsin but it was slowly digested by protease V8. It showed a continuous trimming where no large-limit polypeptides were noticeable thus supporting a monodomain structure. Former appearing peptides have been assigned theoretically according to the protein sequence and predictive methods of accessible areas. Kinetic parameters for the hydrolysis of barley \-glucan and lichenan by measuring the net release of reducing sugars at the optimum pH (7.02) and temperature (55° C) are k
cat=3500 ±800 s–1 (turnover number) and K
m=1.45±0.21 mg/ml for barley \-glucan and k
cat=3000±750 s–1 and K
m=1.98±0.40 mg/ml for lichenan.
Correspondence to: E. Querol 相似文献
8.
Romain Minebois Roberto Pérez-Torrado Amparo Querol 《Environmental microbiology》2020,22(9):3700-3721
The monitoring of fermentation at low temperatures (12–15°C) is a current practice in the winery for retention and enhancement of the flavour volatile content of wines. Among Saccharomyces species, Saccharomyces uvarum and Saccharomyces kudriavzevii have revealed interesting industrial properties, including better adaptation at low temperatures. To gather deeper knowledge of the fermentative metabolism at a low temperature of these species together with S. cerevisiae, we performed a comparative metabolomic analysis using four representative strains. We used batch cultures to obtain an exhaustive and dynamic image of the metabolome of strains passing through the sequential stresses related to the winemaking environment. A great variety of intra- and extracellular metabolites (>500 compounds) were quantified across fermentation using distinct chromatographic methods. Besides a global decrease in the lipid composition of the four strains when they entered into the stationary phase, we reported some strain-specific high magnitude changes. Examples of these differences included divergent patterns of production of short-chain fatty acids and erythritol in the S. uvarum strain. Strains also differed in expression for aromatic amino acid biosynthesis and sulphur metabolism, including the glutathione pathway. These data will allow us to refine and obtain the most value of fermentations with this alternative Saccharomyces species. 相似文献
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