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。     

Effect of phenolic acids on glucose and organic acid metabolism by lactic acid bacteria from wine

The influence of phenolic (p-coumaric, caffeic, ferulic, gallic and protocatechuic) acids on glucose and organic acid metabolism by two strains of wine lactic acid bacteria (Oenococcus oeni VF and Lactobacillus hilgardii 5) was investigated. Cultures were grown in modified MRS medium supplemented with different phenolic acids. Cellular growth was monitored and metabolite concentrations were determined by HPLC-RI. Despite the strong inhibitory effect of most tested phenolic acids on the growth of O. oeni VF, the malolactic activity of this strain was not considerably affected by these compounds. While less affected in its growth, the capacity of L. hilgardii 5 to degrade malic acid was clearly diminished. Except for gallic acid, the addition of phenolic acids delayed the metabolism of glucose and citric acid in both strains tested. It was also found that the presence of hydroxycinnamic acids (p-coumaric, caffeic and ferulic) increased the yield of lactic and acetic acid production from glucose by O. oeni VF and not by L. hilgardii 5. The results show that important oenological characteristics of wine lactic acid bacteria, such as the malolactic activity and the production of volatile organic acids, may be differently affected by the presence of phenolic acids, depending on the bacterial species or strain.     

Ethyl carbamate production by selected yeasts and lactic acid bacteria in red wine

This study was designed to investigate the formation of ethyl carbamate during the fermentation of musts of Vitis vinifera L. cv. Tempranillo and Cabernet Sauvignon by selected yeasts in different conditions of temperature and pH. Secondary malolactic fermentation was then induced by selected lactic acid bacteria. The results indicate an increased concentration of ethyl carbamate after malolactic fermentation, irrespective of the bacteria used or the prevailing physicochemical conditions.     

Species diversity,community dynamics,and metabolite kinetics of spontaneous leek fermentations

Leek (Allium ampeloprasum var. porrum) is one of Belgium's most important vegetables. All or part of the green leek parts are often left on the fields because of their limited cooking applications compared to the white leek parts. Therefore, the possibility to perform leek fermentations in view of product valorization and diversification was investigated. This study deals with the community dynamics, species diversity, and metabolite kinetics of spontaneous leek fermentations, thereby studying the influence of added NaCl concentration, harvesting season, and duration of the fermentation. The combination of a culture-dependent and culture-independent approach revealed the prevalence of lactic acid bacteria (LAB) from the third day of fermentation onwards, which was not influenced by the fermentation conditions applied. Enterobacteriaceae, Pseudomonadaceae, and yeasts disappeared after one week of fermentation. Leuconostoc mesenteroides, Lactobacillus sakei, and Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus parabrevis were the most frequently isolated LAB species. Both added NaCl concentrations were suitable to perform successful fermentations within three weeks. By that time, glucose and fructose, the main leek carbohydrates, were metabolized into mainly lactic acid, acetic acid, ethanol, and mannitol. A sensory analysis revealed that the fermented white leek parts were generally more appreciated than the fermented green leek parts.     

Interactions between yeast, oxygen and polyphenols during alcoholic fermentations: Practical implications

During alcoholic fermentation, even when Saccharomyces cerevisiae cells have used the required oxygen for lipid synthesis, they can consume much more oxygen with no detrimental effect on the fermentation process. Under these conditions, most of the superfluous oxygen is consumed by yeasts by the partial functioning of several nonrespiratory oxygen consumption pathways, which are characterized by the production of reactive oxygen species (ROS). When excess oxygen is added to yeast cells, cell sterol content decreases, following the strong oxidation of intracellular sterols. During aging of fermented products in the presence of nonviable yeast lees (harvested at the end of alcoholic fermentation), the lees can consume oxygen for at least 3 years of the aging process. This oxygen consumption by yeast lees is related to moderate oxidation of yeast membrane lipids by the action of free radicals, strongly decreasing sterols in the yeast lees. The biochemical reactions involved in the oxygen consumption pathways during alcoholic fermentation may be the same as those responsible for oxygen consumption observed in yeast lees. Therefore, the cross-reactivity of complex plant polyphenols, tannins and yeast towards oxygen can easily occur during technological processes (alcoholic fermentation and wine aging). The micro-oxygenation of yeasts releases ROS during alcoholic fermentation and may favour the oxidation of wine phenolic compounds. As yeasts have much higher affinities for oxygen than plant polyphenols, viable yeast and yeast lees compete with phenolic compounds and then hinder the wine aging process. Also, the partial adsorption of plant polyphenols on yeast occurs during alcoholic fermentation, which modifies the overall reactivity of yeast and polyphenols towards oxygen.     

Production of Garcinia wine: changes in biochemical parameters,organic acids and free sugars during fermentation of Garcinia must

Garcinia wine was prepared by fermentation of ameliorated must of Garcinia xanthochymus using Saccharomyces cerevisiae. The present studies focused on changes in biochemical parameters (brix, pH, aldehydes, esters and alcohols), organic acids (reduction of oxalic acid), free sugars and antioxidant activities on fermentation of Garcinia must. The wine had higher amount of residual sugars contributing to the calorific value. The aldehydes and esters content in the final wine were 0.034% and 0.26%, respectively. There was reduction of citric acid and oxalic acid (antinutritional factor) and synthesis of aspartic acid and glutamic acid. Garcinia beverage was accepted on sensory analysis with high score for desirable attributes and overall quality with alcohol content of 6.1%. There was increase in total phenolics (0.039% gallic acid equivalent) and reducing power on fermentation but decrease in 2,2‐diphenyl‐1‐picrylhydrazyl radical scavenging activity.     

Effects of pectinolytic yeast on the microbial composition and spoilage of olive fermentations

This study resulted in the identification of pectinolytic yeasts in directly brined Sicilian-style green olive fermentations and examination of the influence of those yeasts on the microbial composition and quality of fermented olives. Firstly, defective olives processed in Northern California from 2007 to 2008 and characterized by high levels of mesocarp tissue degradation were found to contain distinct yeast and bacterial populations according to DNA sequence-based analyses. Strains of (pectinolytic) Saccharomyces cerevisiae, Pichia manshurica, Pichia kudriavzevii, and Candida boidinii isolated from directly brined olives were then inoculated into laboratory-scale olive fermentations to quantify the effects of individual yeast strains on the olives. The pH, titratable acidity, and numbers of lactic acid bacteria (LAB) and yeasts varied between the fermentations and fermentations inoculated with P. kudriavzevii and C. boidinii promoted the development of LAB populations. Olive tissue structural integrity declined significantly within 30, 74, and 192 days after the inoculation of pectinolytic S. cerevisiae, P. manshurica and C. boidinii, respectively. In comparison, tissue integrity of olives in control fermentations remained intact although pectinolytic yeasts were present. Notably, pectinolytic yeasts were not found in fermentations inoculated with (non-pectinolytic) P. kudriavzevii and olives exposed to a 1:1 ratio of P. kudriavzevii and P. manshurica exhibited no significant tissue defects. This study showed that pectinolytic yeast are important components of directly brined green olive fermentations and damage caused by pectinolytic yeasts might be prevented by other microbial colonists of the olives.     

Fermented whey beverages: A review of process fundamentals,recent developments and nutritional potential

Fermented beverage production from whey is an attractive possibility as whey is a rich source of water and nutrients, beneficial for microbial growth. However, rapid microbial spoilage, scaling of processing equipment, flavour defects and instability of the final product make the production process challenging. Therefore, this study reviews the fundamentals of fermented whey beverage production, while giving specific emphasis to fermentation strategies and process modifications (e.g. membrane separation, additive incorporation and nonthermal pasteurisation) that enhance product quality and shelf life. The review also discusses the nutritional potential of the beverage, imparted by probiotic bacteria, macro and micronutrients and their derivatives.     

Occurrence of biogenic amine-forming lactic acid bacteria in wine and cider

A collection of 810 lactic acid bacteria (LAB) strains isolated from wine and cider was screened for potential biogenic amine (BA) producers by combining molecular and phenotypic approaches. A newly developed multiplex PCR method allowed for the simultaneous detection of four genes involved in the production of histamine (histidine decarboxylase, hdc), tyramine (tyrosine decarboxylase, tyrdc) and putrescine (via either ornithine decarboxylase, odc, or agmatine deiminase, agdi) while TLC and HPLC analysis allowed for BA-production determination. One hundred and fifty-eight LAB strains were monitored by the molecular/phenotypic double approach and revealed a good correlation between genotypic and phenotypic data. Eighteen per cent of the tested strains were positive for at least one BA target gene with up to three detected simultaneously, in particular amongst Lactobacillus brevis and Lactobacillus hilgardii isolates for the tyrdc and agdi genes. The most frequent gene corresponded to the agdi gene detected in 112 strains (14% of all LAB strains) of 10 different LAB species. The tyrdc gene was detected in 67 strains represented by 7 different LAB species (8% overall), especially those isolated from wine. Lower levels of hdc⁺ (2% of strains) and especially odc⁺ (0.5% of strains) strains were observed. Interestingly, species that have never been described to carry BA-producing pathway genes were identified in this study. Furthermore, only one cadaverine-producer was detected and corresponded to Lactobacillus 30a, a collection strain not found in fermented beverages, although cadaverine is commonly detected in wines.     

Hanseniaspora opuntiae, Saccharomyces cerevisiae, Lactobacillus fermentum, and Acetobacter pasteurianus predominate during well-performed Malaysian cocoa bean box fermentations,underlining the importance of these microbial species for a successful cocoa bean fermentation process

Two spontaneous Malaysian cocoa bean box fermentations (one farm, two plantation plots) were investigated. Physical parameters, microbial community dynamics, yeast and bacterial species diversity [mainly lactic acid bacteria (LAB) and acetic acid bacteria (AAB)], and metabolite kinetics were monitored, and chocolates were produced from the respective fermented dry cocoa beans. Similar microbial growth and metabolite profiles were obtained for the two fermentations. Low concentrations of citric acid were found in the fresh pulp, revealing low acidity of the raw material. The main end-products of the catabolism of the pulp substrates glucose, fructose, and citric acid by yeasts, LAB, and AAB were ethanol, lactic acid, acetic acid, and/or mannitol. Hanseniaspora opuntiae, Lactobacillus fermentum, and Acetobacter pasteurianus were the prevalent species of the two fermentations. Saccharomyces cerevisiae, Lactobacillus plantarum, Lactobacillus pentosus, and Acetobacter ghanensis were also found during the mid-phase of the fermentation processes. Leuconostoc pseudomesenteroides and Acetobacter senegalensis were among the prevailing species during the initial phase of the fermentations. Tatumella saanichensis and Enterobacter sp. were present in the beginning of the fermentations and they could be responsible for the degradation of citric acid and/or the production of gluconic acid and lactic acid, respectively. The presence of facultative heterofermentative LAB during the fermentations caused a high production of lactic acid. Finally, as these fermentations were carried out with high-quality raw material and were characterised by a restricted microbial species diversity, resulting in successfully fermented dry cocoa beans and good chocolates produced thereof, it is likely that the prevailing species H. opuntiae, S. cerevisiae, Lb. fermentum, and A. pasteurianus were responsible for it.     

Assessment of the genetic polymorphism and biogenic amine production of indigenous Oenococcus oeni strains isolated from Greek red wines

In the warm climate country of Greece malolactic fermentation (MLF) has received limited attention. Molecular techniques and High Performance Liquid Chromatography (HPLC) were used to study the genetic polymorphism of autochthonous lactic acid bacteria developing towards the end of spontaneous MLF of Greek red wines and for the assessment of their potential to produce harmful biogenic amines. This research revealed that native Oenococcus oeni isolates are very much adapted to specific winery conditions since the majority of spontaneous MLF were driven mostly or exclusively by a single strain of O. oeni. Native O. oeni strains showed only limited dispersion since cluster analysis uncovered only few common genotypes among indigenous isolates from different wineries. The genotype of a frequently used malolactic starter was more than often detected among autochthonous isolates without nevertheless compromising the biodiversity of natural microflora residing in wineries but rather becoming a part of it. For the majority of the wine samples studied, MLF implementation and storage in bottles resulted in negligible changes on the levels of the BA histamine, tyramine, phenylethylamine, cadaverine as well as of ethylamine, methylamine, isobutylamine. We provide evidence that autochthonous O. oeni isolates can only contribute to putrescine accumulation in Greek wines but still the specific trait behaves as strain-specific with a limited dispersion.     

Isolation and characterization of the microbial population of different South African kefir grains

Kefir, a slightly acidic fermented milk, is produced by adding lactic acid bacteria and yeasts, in the form of grains, to milk. The bacteria and yeasts present in the kefir grains are known to vary widely. Selective growth media and morphological and biochemical characteristics were used for the isolation and identification of the microbes present in the grains from eight different sources in South Africa. The kefir grains were activated in milk for only 24 h to prevent any changes in the microbial population of the grains. The microbial numbers varied between 6.4 × 10⁴ and 8.5 × 10 ⁸  cfu/g on the media selective for the bacterial species and between 1.5 × 10 ⁵ and 3.7 × 10 ⁸   cfu/g on the media selective for the yeast species. The bacterial genera that were identified included Lactobacillus , Leuconostoc and Lactococcus and the yeast genera included Zygosaccharomyces , Candida and Saccharomyces . The distribution frequencies of the microbes in the different grains were determined and most of the grains were dominated by two microbial species. No pediococci, acetic acid bacteria or propionibacteria were detected.     

Protection mechanisms of sugars during different stages of preparation process of dried lactic acid starter cultures

Sugars are recognized protectants used in the preparation of dried starter cultures for fermented food industries, particularly as additives for the drying media. They increase viability of the starter cultures during drying and storage. This review intends to summarize and discuss their roles in each step of the preparation process. The main topics cover the role of sugars in the induction of compatible solutes and alteration of fermentation metabolites during growing of cells, the reduction of cryo- and thermal injuries and membrane damage during drying, as well as the formation of sugar glass matrices and the prevention of oxidation during storage. In some topics, proposed protective mechanisms together with corresponding inactivation mechanisms have been discussed. The protective hypotheses as such are preferential exclusion, water replacement, hydration force explanation, and vitrification of sugars.     

荔枝汁经乳酸菌发酵后营养品质的变化及贮藏稳定性研究

研究了荔枝汁分别经干酪乳杆菌、保加利亚乳杆菌和肠膜状明串珠菌发酵后及4 ℃贮藏期间乳酸菌活菌数、主要营养品质和色泽的变化。研究表明：三种乳酸菌都能在荔枝汁中很好的生长,发酵12 h后,活菌数均达到最大值（9.0个对数以上）;三种菌对荔枝汁中糖和有机酸的转化和利用有明显差异,其中干酪乳杆菌和保加利亚乳杆菌发酵荔枝汁后产生了高达37.5 g/L的胞外多糖;荔枝汁分别经三种乳酸菌发酵后,多酚和Vc含量均变化不显著,L*值均明显下降（P<0.05）,且色泽变得更白。另外,三种菌发酵荔枝汁在4 ℃贮藏期间,没有明显的后发酵发生,但经过28 d的低温贮藏后,肠膜状明串珠菌的活菌数下降到了7.0对数以下,而干酪乳杆菌和保加利亚乳杆菌仅仅下降了不到0.5个对数。     

Biodiversity of yeasts, lactic acid bacteria and acetic acid bacteria in the fermentation of "Shanxi aged vinegar", a traditional Chinese vinegar

Shanxi aged vinegar is a famous traditional Chinese vinegar made from several kinds of cereal by spontaneous solid-state fermentation techniques. In order to get a comprehensive understanding of culturable microorganism’s diversity present in its fermentation, the indigenous microorganisms including 47 yeast isolates, 28 lactic acid bacteria isolates and 58 acetic acid bacteria isolates were recovered in different fermenting time and characterized based on a combination of phenotypic and genotypic approaches including inter-delta/PCR, PCR-RFLP, ERIC/PCR analysis, as well as 16S rRNA and 26S rRNA partial gene sequencing. In the alcoholic fermentation, the dominant yeast species Saccharomyces (S.) cerevisiae (96%) exhibited low phenotypic and genotypic diversity among the isolates, while Lactobacillus (Lb.) fermentum together with Lb. plantarum, Lb. buchneri, Lb. casei, Pediococcus (P.) acidilactici, P. pentosaceus and Weissella confusa were predominated in the bacterial population at the same stage. Acetobacter (A.) pasteurianus showing great variety both in genotypic and phenotypic tests was the dominant species (76%) in the acetic acid fermentation stage, while the other acetic acid bacteria species including A. senegalensis, A. indonesiensis, A. malorum and A. orientalis, as well as Gluconobacter (G.) oxydans were detected at initial point of alcoholic and acetic acid fermentation stage respectively.     

龙眼干白葡萄酒发酵过程中生物胺和氨基酸含量的变化

本研究利用高效液相色谱技术分析了龙眼干白葡萄酒酒精发酵和苹乳发酵过程中8种生物胺和22种氨基酸含量的变化。结果表明,在酒精发酵过程中8种生物胺含量均很低,而在苹乳发酵过程中迅速增加。发酵结束时干白葡萄酒中亚精胺和乙醇胺含量最高,其次为组胺和羟色胺。这4种生物胺的前体氨基酸在苹乳发酵阶段含量没有明显变化,而其它大部分氨基酸在酒精发酵过程中含量下降,在苹乳发酵过程中含量呈现不同程度升高。因此,我们认为：葡萄酒发酵过程中氨基酸含量变化与其对应生物胺的变化之间没有相关性,葡萄酒中生物胺的组成比例可能主要取决于乳酸菌中氨基酸脱羧酶的种类和活性。