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SAVE UP TO 15 % WITH CODE MEXICODirected evolution of trypsin for improvement of post-harvest wine quality.
The effect of genetic modification on the composition of the grape was studied using a detailed proteomic approach. The aim was to identify and quantify peptides involved in the grape-wine interaction and the ability of post-harvest wine-making processes to influence their stability. For that, the proteome was analysed after yeast autolysis and after sequential application of fermenting (of a yeast-ethanol solution) and drying (with use of the grapes). This was followed by size-exclusion chromatography (SEC) analysis, liquid chromatography-mass spectrometry (LC-MS) and fast atom bombardment mass spectrometry (FABMS) studies. Finally, the influence of a genetic modification was studied by complementation of the deletion mutant (yeast K1) of trp1, the gene encoding trypsin, by the corresponding wild-type gene (trp1) from Saccharomyces cerevisiae. The wild-type proteome and the yeast autolysate (YAL) were strongly influenced by the fermentation process. The range of peptides identified in YAL, LC-MS and SEC-FABMS data was from 2 to 20 kDa. The degradation of peptides in fermenting grapes was affected by several proteases and peptidases. Enzymatic profiling by HPLC showed that the proteases were in a good proportion, even in the dry grape, which is a finding that supports the regulatory mechanisms that normally occur in fermented wines. Trp1 was successfully expressed in the deletion mutant K1, and fermentation in a medium containing glucose (10 g L(-1)) resulted in a good protease activity of trp1.Limitations of the IL-6/sIL-6R system in post-transplant periodicity: A summary of acute rejection and interstitial damage findings.
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