葡萄酒中生物胺的产生与工艺控制

生物胺存在于多种发酵食品中，人体吸收过量的生物胺后会引起不良的生理反应。在葡萄酒苹果酸—乳酸发酵(MLF)过程中，有些乳酸菌能够对氨基波脱投产生生物胺。利用PCR与DNA探针技术能够快速检测葡萄酒中的组胺产生菌。工艺上采用接种法进行MLF，并在MLF完成后对乳酸菌进行有效清除、可以显著降低葡萄酒中生物胺的含量。     

葡萄酒中的生物胺的生产与工艺控制

生物胺存在于多种发酵食品中，人体吸收过量的生物胺后会引起不良的生理反应，在葡萄酒苹果酸－乳酸发酵（MLF）过程中，有些乳酸菌能够对氨基酸脱羧产生生物胺。利用PCR与DNA探针技术能够快速检测葡萄酒中的组胺产生菌。工艺上采用接种进行MLF，并在MLF完成后对乳酸菌进行有效清除，可以显著降低葡萄酒中生物胺的含量。     

Biogenic amines and the importance of starter cultures for malolactic fermentation

Biogenic amines are compounds produced mainly by microorganisms from the decarboxylation of some amino acids which may cause several health issues in susceptible people. In the winemaking process, one of the factors with the greatest impact on the final concentration of biogenic amines in wine is the malolactic fermentation, which mainly involves the action of lactic acid bacteria. This article gathers scientific literature on the importance of the use of inoculation in winemaking, especially for the potential formation of biogenic amines in spontaneous fermentations, with emphasis on malolactic fermentation. It establishes the set of attributes that influence the formation of biogenic amines in winemaking and aspects related to their toxicity, especially histamine and tyramine. With the growing interest of the market in products with less chemical appeal, the safety of wines, whether from non‐conventional or traditional winemaking, must increasingly be observed. Therefore, the importance of using cultivated starter cultures is discussed, along with the safety of organic wines and the presentation of articles that seek to reconcile the production of organic wine with the preservation of the ‘microbiological terroir’ in a safe way, using selected wild yeasts and lactic acid bacteria.     

Reverse phase HPLC/DAD determination of biogenic amines as dansyl derivatives in experimental red wines

An accurate, rapid and very sensitive high-performance liquid chromatographic (HPLC) method for the determination of dansyl derivative of 11 biogenic amines, in experimental red wine samples from the crop year 2000 produced by Sicilian and French viticultures, was developed. The method involved the derivatisation with dansyl chloride followed by liquid chromatography gradient elution analysis without any other sample pre-treatment. The chromatographic system was equipped with a reversed-phase C-18 column and a DAD detector. A mobile phase of acetonitrile/water was used. The levels of biogenic amines in the samples ranged from 0.10 to 0.80 μg/ml. The HPLC method developed, showed a good linearity, sensitivity and repeatability. Recovery test ranged from 85.0% to 98.6% while detection limit was 0.01 ng/ml for all the studied compounds.     

Influence of technological practices on biogenic amine contents in red wines

To investigate the enological factors associated with biogenic amine content in wines, 224 samples of red wine were industrially manufactured. Differences in the amines and corresponding precursor amino acids of these wines due to their producing cellar and vintage were examined. The main effects of some technological procedures in the biogenic amine content of these wines was also described using multifactor analysis of variance. The results of the study indicate that vintage can clearly influence the amine contents of wines. It was also noticed that some enological practices widely used to enhance wine quality, such as the ageing of wine on lees and, mainly, longer grape skin maceration, strongly increased biogenic amine concentration. However, the addition of pectolytic enzymes did not favour the accumulation of any biogenic amine. With this study, it was also possible to conclude that the inoculation of wine with commercial malolactic starters minimises the levels of biogenic amines.     

Free amino acids and biogenic amines in Alicante Monastrell wines

The simultaneous determination of 17 free amino acids and 8 biogenic amines in Alicante Monastrell wines was investigated for the first time. The quantification was carried out by using a RP-HPLC method, based on a pre-column derivatization with o-phthaldialdehyde (OPA) and fluorescence detection. From the results obtained it may be concluded that the most abundant free amino acids were Glu, Arg, Ala Asp, and Lys. None of the wine samples analysed had histamine (HIM) or Tyramine (TYM) levels above the limits considered as a possible toxic risk for healthy individuals. No measurable amounts of cadaverine (CAD) or methylamine (MEA) were found, showing no spoilage symptoms of sensory properties of the wines. Tryptamine (TRM) content was significantly higher in aged wines compared to young wines. However ethanolamine (ETA) content was lower. These data were used to make a preliminary classification of the samples using cluster analysis.     

苹果酸-乳酸发酵接种研究

本文以四个酿酒葡萄品种和酒类酒球菌(Oenococcus oeni)31DH为材料，进行了苹果酸-乳酸发酵(MLF)接种研究。结果表明，MLF应在酒精发酵结束后接种，接种量以10^7cfu／ml为宜，品种间MLF的速度受接种量的影响。适宜的接种环境条件为：温度18-21℃，pH不低于3．2，总SO2浓度不超过50mg／L。本文还对MLF接种时间的确定和接种量选择问题进行了探讨。     

Biogenic amines content as a measure of the quality of wines of Abruzzo (Italy)

Aim of this research was to study the effect of some agronomic and oenological factors on the content of biogenic amines as quality index of sixty-five Abruzzo wines. Sum of amines was found to be decreasing in the order: red (19.3 ± 12.8 mg L⁻¹), rosé (9.20 ± 6.34 mg L⁻¹), white (7.67 ± 3.84 mg L⁻¹) wine. Significant differences in relationship among amines levels and chemical and chemico-physical characteristics of red, white and rosé wine are due to their different biotechnological process and winemaking. Besides the aging treatment, influential seems to be the effect of the winery, regardless of the area in which it is situated.     

Biogenic amines in wines: Influence of oenological factors

Biogenic amines formation results from the decarboxylation of the corresponding amino acids by action of microorganisms. The presence of these compounds is considered by some authors a fundamental parameter for detriment of alcoholic beverages. The aim of this work was to assay the effect of some oenological factors (viticulture region, grape variety, anti-fungi treatment of grapes, fermentation activators, malolactic starters and storage on lees) from the point of view of their influence on the biogenic amines content of wines. According to our results, it was possible to show that the viticulture region affects the amounts of amines, since wines of some regions present higher contents of amines than wines from other regions. Grape varieties appear to influence the wine amines content. Commercial malolactic starters, after careful selection, should be added to the vinification process in order to decrease the formation of biogenic amines, since in our assays the wines that were inoculated with starters present lower amounts of biogenic amines. The wine storage on lees contributes for a biogenic amines increase.     

Effects of starter cultures and nitrite levels on formation of biogenic amines in sucuk

The effects of starter cultures, starter A (Lactobacillus sakei+Staphylococcus carnosus) and starter B (Pediococcus acidilactici+Staphylococcus xylosus+Lactobacillus curvatus), nitrite levels (0, 75, and 150ppm) and ripening period on biogenic amine production were investigated in sucuk (Turkish dry-fermented sausage) production. Changes in biogenic amines, pH, water activity, moisture, non-protein nitrogen, nitrite and counts of lactic acid bacteria, Micrococcus/Staphylococcus, Enterobacteriaceae, and moulds-yeasts were determined. Ripening period had a very significant effect (P<0.01) on the all biogenic amines. Sausages produced by fermentation with starter cultures, as compared to natural fermentation (control), had lower amounts of putrescine, cadaverine and tyramine (P<0.05). Starter cultures A and B were found to have almost the same effect on the formation of biogenic amines. Use of nitrite in sucuk production affected the formation of biogenic amines except for spermidine and spermine (P<0.01). Lactic acid bacteria counts in the control group were lower than that of starter culture groups. Lactic acid bacteria, Micrococcus/Staphylococcus, and moulds-yeasts counts increased during ripening period while levels of species of Enterobacteriaceae decreased, which was also found to be under the detectable level (<100cfug(-1)), by day 3 of ripening.     

Changes of biogenic amines in Chinese low‐salt fermented fish pieces (Suan yu) inoculated with mixed starter cultures

In this study, two groups of mixed starter cultures (S1, S2), which were isolated from Suan yu, were inoculated for this experimental product, and a batch without starters was used as the control. The effect of the mixed starter cultures on the suppression of biogenic amine (BA) produced during the ripening of Suan yu was investigated. Changes in BA, pH, total volatile base nitrogen (TVB‐N), microbial counts and free amino acids (FAA) contents were detected during ripening. The results show that the mixed starter cultures rapidly decreased pH, inhibited the growth of contaminant microorganisms emerged in the sample and drastically reduced the accumulations of tyramine, putrescine and cadaverine. However, histamine and spermine were not produced in any batch. Besides, pH, FAA and BA production were not directly correlated. The study suggests that inoculation with S1 and S2 inhibited BA, thereby improving the product safety.     

Influence of pH and ethanol on malolactic fermentation and volatile aroma compound composition in white wines

The present study investigated the influences of pH and ethanol on malolactic fermentation (MLF) and the volatile aroma profile of the subsequent white wines from Riesling and Chardonnay inoculated with two different Oenococcus oeni strains. In all cases MLF was induced after completion of alcoholic fermentation (AF). Partial MLF occurred under low pH 3.2 and high alcohol (118.3 g/L) conditions. In the cases with complete MLF, the time required for each strain varied from 13 to 61 days and was dependent on bacterial culture, cultivar and wine parameter. Chemical properties of each wine were determined after AF, complete and partial MLF. The wines showed significant differences in total higher alcohols, esters and acids that are important for the sensory profile and quality of wine. This work demonstrated that the wine matrix as well as the pH and alcohol concentration affects MLF and the final volatile aroma profile. Results indicate that changes in volatile aroma composition are not necessarily related to complete MLF and that partial MLF already has distinct influences on the wine aroma profile of white wines.     

Effect of fermentation with mixed starter cultures on biogenic amines in bighead carp surimi

The effects of one group mixed starter cultures, combined with Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka, and a batch without starter as control on biogenic amines accumulation in the bighead carp surimi during fermentation were investigated. Determination of five different biogenic amines was carried out by reverse‐phase high‐performance liquid chromatography with UV detection. Results showed that mixed starter cultures decreased the pH quickly from initial pH value of 6.4 to 5.2, inhibited the growth of contaminant microorganisms, such as Enterobacteriaceae and Pseudomonas, present in the raw materials, and suppressed the accumulation of histamine, tyramines, spermine and spermidine. Fermentation inoculated with combination of Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka with negative‐decarboxylase activity may prevent biogenic amine formation in the fermented ssurimi and improve hygienic quality of fermented surimi.     

Novel starter cultures to inhibit biogenic amines accumulation during fish sauce fermentation

Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.     

Determination of biogenic amines in Greek wines by HPLC and ultraviolet detection after dansylation and examination of factors affecting their presence and concentration

The presence of biogenic amines in wine has been associated with a number of undesirable physiological effects. The typical levels of 10 biogenic amines in Greek wines were investigated for the first time. One hundred wine samples, varying in type, colour and origin, were analyzed by reversed-phase high performance liquid chromatography (HPLC) and ultraviolet detection after pre-column derivatization with dansyl chloride. The amino acid and organic acid contents of these wines were also evaluated by HPLC in an attempt to explain amine presence and concentration in wines. The total amine average level was 4.76 mg/l. Putrescine and ethylamine were the most prevalent amines, followed by cadaverine and methylamine. Histamine was found in 54.5% of the samples, though only 5.9% of them contained more than 2 mg/l. The levels of histamine and tyramine that were detected are below the amounts considered to have an adverse effect on human health. As regards the amine levels in relation to the different types of tested wines, relatively higher histamine and methylamine contents were detected in sweet wines. High biogenic amines levels were also found in wines from certain grape varieties. The presence of high amine amounts coincided with high acetic and lactic acid levels, which suggests that the biogenic amine formation is related to the microflora of the secondary fermentations, such as bacterial alterations (i.e. metabolization of sugars, citric acid). Some connection between age of wine, reducing sugars and biogenic amine content was also found.     

Comparative study of the amine concentration in wines obtained from the traditional fermentation and from a more anaerobic fermentation method

Content of amines in wine is very variable. Its concentration depends on numerous factors such as raw materials, winemaking conditions, amino acid concentration in must… all these factors are usually interrelated and consequently it is difficult to know the individual action of each one of them. The new vinification method called Ganimede uses a deposit, which utilizes carbonic gas to achieve a more effective maceration of the skins; the conditions inside of the Ganimede tanks are more reductive than inside the conventional tanks. In this study a comparison was made of the amine concentration of wines from Syrah, Merlot and Cabernet Sauvignon musts using the traditional winemaking method (control method) and the Ganimede vinification method. From the results obtained it may be concluded that the Ganimede method produced a greater total concentration of biogenic amines than the conventional method. In general, the concentration of biogenic amines, such as histamine and tyramine, in all three varieties was higher in the wine made by the Ganimede method than in the wine made by the traditional method. These same results were also observed for the volatile amines whose total concentration was also higher in the wine obtained by the Ganimede method than in the control wine.     

Occurrence of biogenic amines in wine: The role of grapes

The evolution of biogenic amines from must to wine has been studied in seven different grape cultivars before and after malolactic fermentation. Alcoholic and malolactic fermentations have been carried out using selected yeasts and bacteria that, in a previous study, were unable to produce biogenic amines. The study has been performed under aseptic conditions to exclude possible interferences due to uncontrolled contaminating microorganisms present in grapes and/or in the environment. The goal of this work was to investigate the influence of grape on biogenic amines content of the wine. The results obtained showed that grape variety is related to the presence of some biogenic amines in wines and that, climatic conditions also affect the accumulation of these compounds in grapes.     

Bacterial community succession and biogenic amine changes during fermentation of fish-chili paste inoculated with different commercial starter cultures

The microbial dynamics, diversity and succession characterisation during the fermentation of fish-chili paste, with and without starter cultures by high-throughput sequencing, were investigated to identify the relationship between microbial composition and the accumulation of biogenic amines. Results showed that Firmicutes was the predominant phylum, and Lactobacillus and Weissella were the main genera during fermentation process, regardless of the inclusion of starter cultures. Compared with naturally fermented samples (NS; no starter cultures), Pediococcus and Staphylococcus flourished in samples inoculated with SBM-52 starter culture (Staphylococcus xylosus + Staphylococcus carnosus + Pediococcus pentosaceus + Pediococcus acidilactici). Staphylococcus also accounted for a larger proportion in samples inoculated with WBX-43 starter culture (Staphylococcus xylosus + Staphylococcus carnosus). Furthermore, Enterobacter was inhibited in inoculated samples. Unweighted pair-group method with arithmetic means and principal component analyses revealed that microbiota structures were different among NS, SBM and WBX samples. In case of biogenic amines, the concentration of putrescine, cadaverine and tyramine increased to higher levels in NS samples compared with inoculated samples. Our work also investigated the relationship between core bacterial communities and biogenic amines. This study provided an improved understanding for the effects of starter cultures on the microbial community and the quality of fish-chili paste.     

Effect of micro-oxygenation on the evolution of aromatic compounds in wines: Malolactic fermentation and ageing in wood

The effect of micro-oxygenation on the aromas of two varietal wines (Tempranillo and Cabernet Sauvignon) when alcoholic fermentation is complete was examined. Several factors, such as whether malolactic fermentation (MLF) took place in barrels or stainless steel vats and ageing time in oak barrels, were taken into account. Major aroma components were studied using GC-FID and minor compounds were studied by GC-MS analysis before and after MLF and after 4 and 8 months of maturation in oak barrels. Sensorial analysis was performed to describe and quantify the different wine notes. Micro-oxygenation affects different compounds but the effect depends to a great extent on the grape varietal used; the effect of micro-oxygenation was more marked on the aroma of Cabernet Sauvignon wine than Tempranillo wines. The concentration of extractable compounds of wood was higher in wines after MLF in barrels than in wines after MLF in steel vats; wines with steel vat MLF had fewer toast and wood notes. The compounds that showed significant differences in concentration after 8 months of maturation were present in higher concentrations in wines fermented in barrels than in stainless steel vats. Barrel-fermented wines were sweeter, with wood and toast notes, fewer alcohol notes and fewer reductive notes.