The aim of this work was to verify the potential of infrared （IR） spectroscopy in near and mid regions to detect the beginning of the malolactic fermentation （MLF） occurring in a model-wine and the further cells a...The aim of this work was to verify the potential of infrared （IR） spectroscopy in near and mid regions to detect the beginning of the malolactic fermentation （MLF） occurring in a model-wine and the further cells autolysis. MLF in wine is a secondary biotransformation due to lactic acid bacteria that usually occurs spontaneou,;ly or after starter inoculation at the end of alcoholic fermentation. Nowadays, it is desirable to supply winemakers with a new rapid and non-destructive approach to monitor MLF progress and 1R spectroscopy technology appears to be suitable for this purpose. The transformation of L-malic acid into L-lactic acid was carried out by inoculating a synthetic wine with an Oenococcus oeni culture and it was monitored through microbiological and chemical methods. At the same time, Fourier transform near infrared （FT-NIR） spectral data, in diffusive transflection mode using an optic probe, and FT-IR spectra, using an germanium crystal attenuated total reflectance （ATR） cell, were collected. Principal component analysis of the spectra was able to identify absorption bands related to the key molecular modifications that took place during the L-malic acid transformation. Thus, the samples were discriminated according to the fermentation phase. Although this study is a preliminary approach, results confirm that near infrared （NIR） and mid infrared （MIR） spectroscopy could be successfully applied to detect the start of MLF and the autolysis of the lactic acid bacteria （LAB） cells.展开更多
Malolactic enzyme is the function enzyme which catalyses the reaction for L-malate converting to L-lactic during malolactic fermentation （MLF）. In this paper, researches concerning the malolactic enzyme gene mleA cl...Malolactic enzyme is the function enzyme which catalyses the reaction for L-malate converting to L-lactic during malolactic fermentation （MLF）. In this paper, researches concerning the malolactic enzyme gene mleA cloned from a patent strain Oenococcus oeni SD-2a screened in Chinese wine and integrated expressing in Saccharomyces cerevisiae were performed in order to carry out both alcoholic fermentation （AF） and malolactic fermentation （MLF） during winemaking, with a view to achieving a better control of MLF in enology. To construct the expression plasmid named pYILmleA, cloned mleA gene, PGK1 promoter, and ADH1 terminator were ligated and inserted into integrating vector YIp5. Yeast transformants were screened on SD/-Ura and identified by auxotrophic test, mating type test, and colony PCR. Target protein was detected by SDS-PAGE and the targeted gene integrated to the chromosome was detected by dot bloting hybridization. After the transformant was cultured in SD/-Ura adding glucose （10%） and L-malate （5 648 mg L-1） for 4 d, the culture supernatant was collected and L-malate and L-lactic acid contents were detected by HPLC. 1 278-1 312 mg L-1 L-lactic acids were detected, while the comparative drop rates of L-malate were 20.18-20.85%. L-malate and L-lactic contents of the transformants showed extra significant difference and significant difference with the control ones by t-test respectively. The result indicated that the functional expression was achieved in recombinants S. cerevisiae.展开更多
The efficiency of using nanostructured silver in technological processes of making red wine has been studied with the purpose of substituting sulfur dioxide. For preparation of research objects, we used dry wine mater...The efficiency of using nanostructured silver in technological processes of making red wine has been studied with the purpose of substituting sulfur dioxide. For preparation of research objects, we used dry wine material of red grape variety ＂Saperavi＂. After completion of malolactic fermentation, the second racking the wine material off the lees and its treatment with antiseptics were carried out. Sulfur dioxide （Kadifit） and different doses of nanostructured silver were used. Microbiological investigation was performed on the presence of lactic bacteria after completion of malolactic fermentation and racking off the lees. The investigation of phenolic compounds was conducted at the following stages of technological processes： （1） after completion of alcoholic fermentation and racking off the lees; （2） after completion of malolactic fermentation; （3） after the second racking off the lees and treatment with antiseptics. By means of the HPLC analysis, the amounts ofcatechins, phenolcarbonic acids and flavonols were determined. It has been established that after conduction of malolactic fermentation in the process of racking the wine material off the lees and storage, the application of 0.6 mg/L of nano-silver for blocking of lactic-acid bacteria and inhibition of oxidation of phenolic compounds has the same effect as sulfitization with 50 mg/L of Kadifit.展开更多
文摘The aim of this work was to verify the potential of infrared （IR） spectroscopy in near and mid regions to detect the beginning of the malolactic fermentation （MLF） occurring in a model-wine and the further cells autolysis. MLF in wine is a secondary biotransformation due to lactic acid bacteria that usually occurs spontaneou,;ly or after starter inoculation at the end of alcoholic fermentation. Nowadays, it is desirable to supply winemakers with a new rapid and non-destructive approach to monitor MLF progress and 1R spectroscopy technology appears to be suitable for this purpose. The transformation of L-malic acid into L-lactic acid was carried out by inoculating a synthetic wine with an Oenococcus oeni culture and it was monitored through microbiological and chemical methods. At the same time, Fourier transform near infrared （FT-NIR） spectral data, in diffusive transflection mode using an optic probe, and FT-IR spectra, using an germanium crystal attenuated total reflectance （ATR） cell, were collected. Principal component analysis of the spectra was able to identify absorption bands related to the key molecular modifications that took place during the L-malic acid transformation. Thus, the samples were discriminated according to the fermentation phase. Although this study is a preliminary approach, results confirm that near infrared （NIR） and mid infrared （MIR） spectroscopy could be successfully applied to detect the start of MLF and the autolysis of the lactic acid bacteria （LAB） cells.
基金supported by the National High-Tech Research and Development Program of China (863 Program of China, 2007AA10Z314)andthe Earmarked Fund for Modern Agro-Industry Technology Research System, China (Z225020801)
文摘Malolactic enzyme is the function enzyme which catalyses the reaction for L-malate converting to L-lactic during malolactic fermentation （MLF）. In this paper, researches concerning the malolactic enzyme gene mleA cloned from a patent strain Oenococcus oeni SD-2a screened in Chinese wine and integrated expressing in Saccharomyces cerevisiae were performed in order to carry out both alcoholic fermentation （AF） and malolactic fermentation （MLF） during winemaking, with a view to achieving a better control of MLF in enology. To construct the expression plasmid named pYILmleA, cloned mleA gene, PGK1 promoter, and ADH1 terminator were ligated and inserted into integrating vector YIp5. Yeast transformants were screened on SD/-Ura and identified by auxotrophic test, mating type test, and colony PCR. Target protein was detected by SDS-PAGE and the targeted gene integrated to the chromosome was detected by dot bloting hybridization. After the transformant was cultured in SD/-Ura adding glucose （10%） and L-malate （5 648 mg L-1） for 4 d, the culture supernatant was collected and L-malate and L-lactic acid contents were detected by HPLC. 1 278-1 312 mg L-1 L-lactic acids were detected, while the comparative drop rates of L-malate were 20.18-20.85%. L-malate and L-lactic contents of the transformants showed extra significant difference and significant difference with the control ones by t-test respectively. The result indicated that the functional expression was achieved in recombinants S. cerevisiae.
文摘The efficiency of using nanostructured silver in technological processes of making red wine has been studied with the purpose of substituting sulfur dioxide. For preparation of research objects, we used dry wine material of red grape variety ＂Saperavi＂. After completion of malolactic fermentation, the second racking the wine material off the lees and its treatment with antiseptics were carried out. Sulfur dioxide （Kadifit） and different doses of nanostructured silver were used. Microbiological investigation was performed on the presence of lactic bacteria after completion of malolactic fermentation and racking off the lees. The investigation of phenolic compounds was conducted at the following stages of technological processes： （1） after completion of alcoholic fermentation and racking off the lees; （2） after completion of malolactic fermentation; （3） after the second racking off the lees and treatment with antiseptics. By means of the HPLC analysis, the amounts ofcatechins, phenolcarbonic acids and flavonols were determined. It has been established that after conduction of malolactic fermentation in the process of racking the wine material off the lees and storage, the application of 0.6 mg/L of nano-silver for blocking of lactic-acid bacteria and inhibition of oxidation of phenolic compounds has the same effect as sulfitization with 50 mg/L of Kadifit.
