Free amino acids and volatile compounds in vinegars obtained from different types of substrate

The present work reports on the free amino acids and volatile compounds in vinegars obtained from different types of raw materials (cider, white wine and red wine). Based on the amino acid contents of three types of vinegar and of the substrates from which they were obtained, the profile of the nitrogen‐supplying compounds and their uptake for the acetic acid bacteria were very similar. The most important amino acid in terms of supply and uptake was found to be L ‐leucine. L ‐Proline proved also important in the wine vinegars, however it was not depleted as L ‐leucine. The different acetification reactions involved were found to yield acetoin, and plus 1,1‐diethoxyethane in the wine vinegars. Copyright © 2004 Society of Chemical Industry     

Improving acetic acid production of Acetobacter pasteurianus AC2005 in hawthorn vinegar fermentation by using beer for seed culture

Fruit vinegar can be obtained biologically from the fermentation of fruit wine using species of the genera Acetobacter. However, the productivity of vinegar is generally low because of the inhibition by substrate ethanol and product acetic acid. In this research, the yield of acetic acid, as well as the cell growth, in hawthorn vinegar fermentation was enhanced using beer for seed preparation. The higher yield of acetic acid of AC2005 was because of the higher alcohol dehydrogenase and aldehyde dehydrogenase activities caused by using beer for seed preparation. Furthermore, Acetobacter pasteurianus AC2005 showed acid stability during the fed‐batch fermentation of hawthorn vinegar.     

Antioxidant activity and phenolic content of wine vinegars produced by two different techniques

BACKGROUND: The presence of phenolics in fruit, red wine and vinegar has positive health effects due to their significant antioxidant activity. The aim of the study was to determine the effects of two different vinegar production methods on antioxidant activity and phenolic level of vinegars derived from Ulugbey Karasi grapes. Traditional surface and industrial submerge methods were used to make vinegar. Samples were taken from fresh red grape juice, maceration, wine, traditional vinegar and industrial vinegar. RESULTS: Total phenolic content of traditional and industrial vinegar samples were 2690 mg L^?1 and 2461 mg L^?1 GAE, respectively. ORAC values of traditional and industrial vinegar samples were 10.50 µmol mL^?1and 8.84 µmol mL^?1 TE, respectively. Antioxidant activity values of traditional and industrial vinegars were 13.50 mmol L^?1 and 10.37 mmol L^?1 TEAC, respectively. Gallic acid, catechin, epicatechin, chlorogenic acid, caffeic acid, syringic acid, p‐coumaric acid and ferulic acid were detected in grape juice, wine and vinegar samples. The content of catechin in industrial vinegar (27.50 mg L^?1) was significantly higher than that of in traditional vinegar (13.76 mg L^?1) (P < 0.05). Traditional vinegar had higher amounts of chlorogenic and syringic acids than the industrial vinegar (P > 0.05). CONCLUSION: Results of this study showed that different production methods affected the functional constituents of wine vinegars. Copyright © 2010 Society of Chemical Industry     

Rapid molecular methods for enumeration and taxonomical identification of acetic acid bacteria responsible for submerged vinegar production

The aim of the present study was to search for a rapid and reliable method to enumerate viable acetic acid bacteria (AAB) and to identify to genera and species level AAB isolates from vinegars in full acetic fermentation elaborated by the submerged method from cider, wine and spirit ethanol in industrial bioreactors. Results showed that the rapid epifluorescence staining method using the LIVE/DEAD BacLight bacterial viability kit and direct counts in Neubauer chamber rendered consistent and reliable data for viable cell counts of bacteria in all the studied vinegars. A linear correlation was shown between viable cell counts and fermentation rates. The highest fermentation rates and viable cell counts were found in cider vinegars, whereas spirit vinegars showed the lowest values for both parameters. Eighty-four AAB pure isolates were recovered from 41 different vinegar samples and were submitted to DNA extraction. PCR amplification of the 16S–23S intergenic spacer region of rDNA and subsequent sequencing were carried out to identify isolates to species level. Results showed that Gluconacetobacter europaeus was the predominant cultivable species, appearing in 79% of the total isolates. This was the unique species found in spirit vinegars, and this is the first time that AAB from spirit vinegars are taxonomically identified. Ga. europaeus was as well the predominant cultivable species in white wine vinegars. Cider vinegars presented the highest variability of species: Ga. europaeus (35.3% appearance among cultivable isolates), Ga. xylinus (35.3%), Acetobacter pasteurianus (17.6%) and Ga. hansenii (11.8%). Red wine vinegars showed cultivable isolates of the species Ga. xylinus (71.4%) and Ga. europaeus (28.6%). Summarising, both described methods for AAB enumeration and taxonomical identification proved to be fast and reliable methods, and results revealed Ga. europaeus as the cultivable major species in vinegars in full fermentation conducted by the submerged method, suggesting that Ga. europaeus strains can constitute excellent starter cultures for the elaboration of vinegars by the submerged method.     

A Note — Production of Vinegar from Whey

Cheese whey supplemented with lactose was employed to produce vinegar. Whey was first transformed into an alcoholic beverage via fermentation with the yeast Kluyveromyces fragilis, and then the alcoholic product was employed as a substrate for an acetic acid fermentation. The bacteria used for the acetic acid production process were isolated from a starter culture provided by a vinegar factory. The bacteria employed were classified as Acetobacter pasteurianus. The vinegar obtained had a concentration of acetic acid between 5 and 6% (v/v). Ethyl acetate and fusel alcohols (isobutanol, 2‐methyl‐1‐butanol and 3‐methyl‐1‐butanol) were detected in the final product. The process and the substrates employed satisfied FAO requirements that the product was acceptable for human consumption. The efficiency of bio‐transformation of ethanol into acetic acid was 84%.     

提高柑桔果醋中烟酸含量的研究

对柑桔果醋发酵过程中烟酸含量的变化进行了研究。通过对不同发酵条件的比较,结果发现烟酸主要是由酵母代谢产生。证明了柑桔清酒中添加适量酒母与酵母膏进行醋酸发酵,所得果醋的烟酸含量最高,达326mg/L。研究了不同发酵条件对烟酸含量的影响,并使烟酸含量提高至390mg/L。所研制的保健果醋中烟酸含量约为市售米醋的3倍,且黄酮含量较高。     

不同醋酸菌及发酵方式对柑橘果醋品质的影响

为了探讨不同醋酸菌对柑橘果醋品质的影响,该研究以柑橘为原料,采用沪酿1.01醋酸菌、AS1.41醋酸菌和许氏醋酸菌三种醋酸菌,通过一次发酵法和二次发酵法酿造六种柑橘果醋,并对这6种果醋的理化指标进行分析。结果表明,采用AS1.41醋酸菌、一次发酵法加工的果醋总多酚、总黄酮、蛋白质、单体酚、总氨基酸（TAA）、必需氨基酸（EAA）、非必需氨基酸（NAA）、呈味氨基酸含量分别为0.49 mg/mL、0.36 mg/mL、4.51 mg/mL、23.49 mg/L、1 830.10 mg/kg、420.80 mg/kg、1 409.30 mg/kg和1 822.10 mg/kg,均高于其他五种果醋,沪酿1.01和AS1.41醋酸菌、一次发酵法有机酸总含量较高,分别为11 786.03 mg/L和10 505.51 mg/L。综上所述,6种柑橘果醋中AS1.41醋酸菌、一次发酵法加工的柑橘果醋品质最好。     

Evaluation and optimisation of bacterial genomic DNA extraction for no-culture techniques applied to vinegars

Direct genomic DNA extraction from vinegars was set up and suitability for PCR assays performed by PCR/DGGE and sequencing of 16S rRNA gene. The method was tested on 12 intermediary products of special vinegars, fruit vinegars and condiments produced from different raw materials and procedures. DNAs extraction was performed on pellets by chemical, enzymatic, resin mediated methods and their modifications. Suitable yield and DNA purity were obtained by modification of a method based on the use of PVP/CTAB to remove polyphenolic components and esopolysaccharides. By sequencing of bands from DGGE gel, Gluconacetobacter europaeus, Acetobacter malorum/cerevisiae and Acetobacter orleanensis were detected as main species in samples having more than 4% of acetic acid content. From samples having no acetic acid content, sequences retrieved from excised bands revealed high similarity with prokaryotes with no function on vinegar fermentation: Burkholderia spp., Cupriavidus spp., Lactococcus lactis and Leuconostoc mesenteroides. The method was suitable to be applied for no-culture study of vinegars containing polyphenols and esopolysaccharides allowing a more complete assessment of vinegar bacteria.     

柑桔醋醋酸菌的筛选研究

经过对几株醋酸菌在柑桔醋发酵过程中的性能进行测试,筛选出了适合柑桔醋发酵的QY7009菌株。结果表明,QY7009菌株在柑桔酒中的产酸速度和乙醇转化率等高于常用的AS1.41和沪酿1.01菌株,在酒度为6°的柑桔酒中,QY7009菌种的乙醇转化率高达87%。     

Determination of major volatile compounds during the production of fruit vinegars by static headspace gas chromatography-mass spectrometry method

A static headspace gas chromatography coupled to mass spectrometry (SHS-GC-MS) method was validated to determine several major volatile components during the production process of fruit vinegars. The method is simple, fast, linear in the working range, suitably sensitive, repeatable and reproducible, and has a good degree of accuracy for most of the compounds studied. Different conditions were tested in the production process of vinegars by means of double fermentation. The addition of SO₂ and pectolytic enzymes produced a considerable increase in methanol and acetaldehyde, especially in strawberry purees, whereas pressing led to a loss of these volatile compounds. In the alcoholic fermentation of persimmon and strawberry purees, the Saccharomyces cerevisiae strain used had a great influence on the production of acetaldehyde and higher alcohols in wines. Considering the influence of these studied compounds in the final profile of the vinegars, our results showed that the S. cerevisiae strain isolated in this study produced the most suitable wine substrates for the production of vinegars. Moreover, semisolid fruit substrate provides better results than liquid substrate. Inoculated acetification in wood recipients yielded vinegars with a better volatile profile, as these contained higher levels of most compounds except acetaldehyde.     

Effect of nuruk and fermentation method on organic acid and volatile compounds in brown rice vinegar

The effects of different nuruk contents and fermentation methods (AV, vinegar fermented in an agitated culture; SV, vinegar fermented in a static culture) on organic acids and volatile compounds in brown rice vinegars were investigated. In the SV, the contents of acetic, oxalic, tartaric, and malic acids increased with hipher contents of nuruk. Acetic, tartaric, and malic acid contents were higher in the SV than those in the AV. Volatile compounds that can affect vinegar quality, including acetic acid, isoamyl acetate, phenethyl acetate, and phenethyl alcohol were present at high concentrations in the AV. With the increase in nuruk contents in the AV, acetic acid content decreased and isoamyl acetate and phenethyl acetate content increased. No significant differences in sensory scores were observed regarding the amount of nuruk and the type of fermentation. However, electronic-nose analysis showed its potential to effectively differentiate different samples.     

Continuous production of wine vinegar in bubble column reactors of up to 60-litre capacity

This paper describes the development of a continuous acetic acid fermentation process for the production of wine vinegar in bubble column reactors of up to 60 l capacity. To determine appropriate fermentation conditions a study of the influence of residual ethanol concentration, inlet flow rate and aeration was carried out using a 6-l laboratory reactor, white table wine as fermentation medium, a temperature of 30 °C and an air flow rate of 0.125 min^-1 (vvm). The concentration of acetic acid obtained in the continuous wine vinegar production ranged from 91 g/l at 28.6 ml/h to 28 g/l at 154.1 ml/h by increasing the inlet flow rate. As expected, the biomass decreased as well, from 208 mg/l to 106 mg/l. The maximum acetification rate was observed in the range 85-110 ml/h, corresponding to a value of about 1.1 g/l/h. A further increase in the flow rate produced a slight decrease in the acetification rate. Best yields, between 94.5 and 94.7%, were obtained in the flow rate range of 60-75 ml/h. The acetification rate was improved only by about 10% by increasing the aeration from 0.125 to 0.250 min^-1. The continuous wine vinegar production was scaled up from the laboratory fermentor to a 60-l pilot acetator. During the steady state (residential time >6), with an inlet flow rate of 950 ml/h, temperature of 30 °C and aeration of 0.250 min^-1, the following parameters were obtained: acetic acid concentration 72 g/l, overall productivity 1.41 g/l/h and yield 94.2%.     

木瓜酒和木瓜醋发酵工艺及其有机酸组成分析

以光皮木瓜为原料,发酵木瓜干酒,再通过液态深层发酵酿制木瓜醋,确定了木瓜干酒和木瓜醋的发酵工艺,并对木瓜酒和醋主要有机酸进行分析。光皮木瓜经榨汁、调整糖度后进入发酵工序,酒精发酵采用带皮渣半固态发酵方式,条件为加水比例1.5∶1（m/m）、初始糖度18%、果酒干酵母接种量0.1%、在24 ℃条件下发酵64 h,木瓜酒酒度（乙醇体积分数）为9.45%。醋酸发酵采用半连续式液态深层发酵法,调整初始酒度7%,醋酸菌接种量10%,在34 ℃条件下醋酸发酵80 h,木瓜醋总酸度为4.52%;分割留种发酵仅需24 h即可完成醋酸发酵。采用反相高效液相色谱法从木瓜干酒和木瓜醋中检出10 种有机酸,分别是草酸、酒石酸、甲酸、苹果酸、α-酮戊二酸、乳酸、醋酸、柠檬酸、富马酸和琥珀酸。实验确定的发酵工艺以及有机酸的分析与鉴定可为木瓜干酒及木瓜醋产品的开发提供理论依据。     

秋香梨果醋醋酸发酵工艺优化及抗氧化活性研究

以秋香梨酒为原料,采用沪酿1.01醋酸菌,以总酸含量为指标,利用响应面法优化果醋醋酸发酵工艺,并评价果醋的抗氧化能力。结果表明:秋香梨果醋醋酸发酵的最佳工艺条件为醋酸菌接种量9.4%,发酵时间6d,发酵温度31℃,初始pH 4.5。此条件下得到的果醋色泽金黄,果香醇厚,其总酸含量为54.45g/L。秋香梨酒经醋酸菌发酵后,其DPPH自由基清除率、ABTS自由基清除率和Fe3+还原能力分别提高了21.17%,23.65%,30.23%,表明该果醋具有良好的抗氧化活性。     

Strain typing of acetic acid bacteria responsible for vinegar production by the submerged elaboration method

Strain typing of 103 acetic acid bacteria isolates from vinegars elaborated by the submerged method from ciders, wines and spirit ethanol, was carried on in this study. Two different molecular methods were utilised: pulsed field gel electrophoresis (PFGE) of total DNA digests with a number of restriction enzymes, and enterobacterial repetitive intergenic consensus (ERIC) – PCR analysis. The comparative study of both methods showed that restriction fragment PFGE of SpeI digests of total DNA was a suitable method for strain typing and for determining which strains were present in vinegar fermentations. Results showed that strains of the species Gluconacetobacter europaeus were the most frequent leader strains of fermentations by the submerged method in the studied vinegars, and among them strain R1 was the predominant one. Results showed as well that mixed populations (at least two different strains) occurred in vinegars from cider and wine, whereas unique strains were found in spirit vinegars, which offered the most stressing conditions for bacterial growth.     

Influence of Fermentation Process on the Anthocyanin Composition of Wine and Vinegar Elaborated from Strawberry

Anthocyanins are the major polyphenolic compounds in strawberry fruit responsible for its color. Due to their sensitivity, they are affected by food processing techniques such as fermentation that alters both their chemical composition and organoleptic properties. This work aims to evaluate the impact of different fermentation processes on individual anthocyanins compounds in strawberry wine and vinegar by UHPLC‐MS/MS Q Exactive analysis. Nineteen, 18, and 14 anthocyanin compounds were identified in the strawberry initial substrate, strawberry wine, and strawberry vinegar, respectively. Four and 8 anthocyanin compounds were tentatively identified with high accuracy for the 1st time to be present in the beverages obtained by alcoholic fermentation and acetic fermentation of strawberry, respectively. Both, the total and the individual anthocyanin concentrations were decreased by both fermentation processes, affecting the alcoholic fermentation to a lesser extent (19%) than the acetic fermentation (91%). Indeed, several changes in color parameters have been assessed. The color of the wine and the vinegar made from strawberry changed during the fermentation process, varying from red to orange color, this fact is directly correlated with the decrease of anthocyanins compounds.     

Anthocyanin composition in Cabernet Sauvignon red wine vinegar obtained by submerged acetification

Vinegars elaborated from white wine can be characterized by their phenolic composition. Indeed, for authenticity purpose, phenolic composition can be used to identify Sherry and Balsamic vinegars. However, the phenolic composition of red wine vinegars has scarcely been studied. Anthocyanin compounds in particular remain largely unknown. This study focuses on the analysis of anthocyanin compounds in red wine vinegar and the effect of acetification with submerged culture on such vinegars.The vinegar used in this study was produced from a young Cabernet Sauvignon wine in a laboratory-scale fermenter. Subsequent analyses of both wine and vinegar included their anthocyanin profile (by LC/MS), and their non-anthocyanin phenolic profile (by LC/DAD). In addition, wine and vinegar anthocyanin extracts were fractionated by CCC to determine the contribution of the fractions to overall antioxidant activity (AA), using ORAC, FRAP and DPPH assays.A total of 20 anthocyanin compounds were identified in the vinegar. As far as we know, this is the first time that anthocyanin-derived pigments (pyranoanthocyanins and ethyl-linked compounds) have been identified in red vinegar in such detail. Moreover, an original contribution of this study is the identification for the first time of catechyl-pyranocyanidin-3-glucoside in vinegar and wine, as well as two anthocyanin compounds not previously reported in vinegar or Cabernet Sauvignon wine: acetyl vitisin B and coumaroyl vitisin B. After the acetification process, vitisin-type and ethyl-linked compounds increased and monomeric anthocyanins, phenolic acids (ferulic acid, caffeic acid and caftaric acid) and flavan-3-ol ((+)-catechin) decreased.Although the proportion by weight of the polymeric compound fraction is similar in wine and vinegar, the AA of these polymers in vinegar is significantly greater (p < 0.05). We have also determined for the first time an approximate value of AA for malvidin-3-(6-acetyl)-glucoside isolated from vinegar.     

不同洁净度葡萄的葡萄酒发酵效果研究

使用武宣地区3种红曲米作为发酵剂酿造红曲柿酒，分析其理化指标和挥发性风味成分差异，并采用Illumina MiSeq高通量测序技术检测3种红曲米的细菌多样性，以解析不同细菌群落在红曲柿酒发酵过程中的作用。结果表明，不同红曲柿酒总酸和挥发酸含量差异显著（P＜0.05）；挥发性酸类物质组成存在较大差异，其中红曲米样品WX-2制成的红曲柿酒挥发性酸类物质含量最高（10.0%）。从3种红曲米样品共检测出19个细菌门，239个细菌属，共有优势菌门为变形菌门（Proteobacteria）、厚壁菌门（Firmicutes）、拟杆菌门（Bacteroidetes）、放线菌门（Actinobacteria）；共有优势菌属为伯克霍尔德菌属（Burkholderia）、克雷伯氏菌属（Klebsiella），WX-2样品中优势菌属还有醋菌属（Acetobacter）、小球菌属（Pediococcus）、乳酸杆菌属（Lactobacillus），均为产酸菌株。表明红曲米细菌群落结构与红曲柿酒酸类物质的形成紧密相关。     

“巴伦比”发酵型无核黄皮果醋的研制

以广东云浮郁南县优质成熟无核黄皮为原料,加入植物提取用酶控温酶解,然后加入葡萄酒酿酒酵母、多微麸曲、活性醋酸菌和玉米芯进行酒精发酵与醋酸发酵同步进行,最后降低温度加入皂土陈酿后过滤,即得到保健功效好的“巴伦比”无核黄皮果醋。该无核黄皮果醋呈棕黄色,具有无核黄皮特有的果香和浓郁的醋酒香,酸味纯正柔和、口味微甘爽口,稳定性好。     

醋酸菌麸曲制备工艺的优化

以巴氏醋杆菌C9-4(Acetobacter pasteurianum strain C9-4)为麸曲发酵菌株,应用响应面法优化了醋酸菌麸曲的制备工艺条件。结果表明,其最佳制备条件为:乙酸添加量1.03 mL/100 g、乙醇添加量2.09 mL/100 g、接种量4.09mL/100g,该条件下醋酸菌菌落总数达到1.38×10~8 CFU/g。应用该条件,在食醋企业对醋酸菌麸曲进行逐级扩大培养,扩培过程中麸曲醋酸菌总数均能达到10~8 CFU/g以上。