Monitoring the microbial community during solid-state acetic acid fermentation of Zhenjiang aromatic vinegar

Zhenjiang aromatic vinegar is one of the most famous Chinese traditional vinegars. In this study, change of the microbial community during its fermentation process was investigated. DGGE results showed that microbial community was comparatively stable, and the diversity has a disciplinary series of changes during the fermentation process. It was suggested that domestication of microbes and unique cycle-inoculation style used in the fermentation of Zhenjiang aromatic vinegar were responsible for comparatively stable of the microbial community. Furthermore, two clone libraries were constructed. The results showed that bacteria presented in the fermentation belonged to genus Lactobacillus, Acetobacter, Gluconacetobacter, Staphylococcus, Enterobacter, Pseudomonas, Flavobacterium and Sinorhizobium, while the fungi were genus Saccharomyces. DGGE combined with clone library analysis was an effective and credible technique for analyzing the microbial community during the fermentation process of Zhenjiang aromatic vinegar. Real-time PCR results suggested that the biomass showed a “system microbes self-domestication” process in the first 5 days, then reached a higher level at the 7th day before gradually decreasing until the fermentation ended at the 20th day. This is the first report to study the changes of microbial community during fermentation process of Chinese traditional solid-state fermentation of vinegar.     

Recovery of aroma compounds from Zhenjiang aromatic vinegar by supercritical fluid extraction

Zhenjiang aromatic vinegar, which is produced from sticky rice through solid‐state fermentation, is highly prized as one of the four famous China‐style vinegars, owing to its unique flavour. In this study, a method of supercritical fluid extraction (SFE) was used for the first time to recover aroma compounds from Zhenjiang aromatic vinegar. For this purpose, the optimal conditions for the extraction of aroma compounds by SFE were determined as follows: CO₂ flow rate, 25 L h^?1; extraction time, 2 h; extraction pressure, 35 MPa; and extraction temperature, 323 K. Using headspace solid‐phase microextraction‐gas chromatography–mass spectrometry (HS‐SPME‐GC‐MS) analysis, a total of 49 and 44 aroma compounds were identified in Zhenjiang aromatic vinegar and its SFE extract, respectively. Acetic acid, ethyl acetate, furfural, phenethyl alcohol, tetramethyl‐pyrazine, 3‐hydroxybutanone and benzaldehyde were the main aroma compounds in the vinegar and its SFE extract. SFE is a fast and sensitive method to recovery aroma compounds from the vinegar.     

食醋生产过程中铁、锌、铜的迁移变化

本文研究了食醋发酵过程中铁、锌、铜的产生途径及影响因素。采用原子吸收分光光度法对镇江香醋不同发酵阶段液相、固相中的铁、锌、铜含量变化进行分析。结果表明铁、锌、铜在发酵各阶段液相中主要以可溶态存在,沉淀的形成对Fe、Zn、Cu具有富集作用。随着发酵的进行,发酵设备、管道由于长期受醋酸、有机酸等的腐蚀使铁、锌、铜游离和在发酵生产过程中添加了含有铁、锌、铜的辅料,是醋醅中铁、锌、铜含量上升的主要因素。醋中铁主要来源于醋酸发酵和封醅陈酿阶段,在淋醋时铁浓度达到最高,59.69 mg/L。醋中的锌、铜主要来源于酒精发酵阶段,浓度分别达到18.12 mg/L和0.60 mg/L。由于醋醅对铁、锌、铜元素具有强吸附力,醋醅对醋液中的金属元素浓度具有一定的调控作用。通过本研究表明改进设备材质、采用合理的过滤措施及在发酵过程中使用金属离子吸附剂将是控制食醋中金属含量的有效措施。     

HS‐SPME/GC‐MS and chemometrics for volatile composition of Chinese traditional aromatic vinegar in the Zhenjiang region

The main purpose of this study was to determine the volatile composition of Zhenjiang aromatic vinegar, one of the four famous China‐style cereal vinegars, by using headspace solid‐phase microextraction (HS‐SPME)/gas chromatography‐mass spectrum (GC‐MS) and chemometrics. For this purpose, the HS‐SPME sampling method for the volatile compounds of Zhenjiang aromatic vinegar was optimized by a second‐order rotatable central composite experimental design (CCD). A HS extraction of the volatile compounds by incubation on a 65 µm thickness polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre during 44.2 min at 69.5°C with 1.9 g NaCl add gave the most effective and accurate extraction. By the optimized method, a total of 58 volatile compounds, including 9 alcohols, 13 acids, 16 esters, 5 aldehydes, 4 ketones and 8 heterocycle compounds, were identified from 13 aromatic vinegar samples manufactured in Zhenjiang region. By principal components analysis (PCA), the thirteen vinegar samples were classified into 3 groups, and 10 volatile compounds were chosen as characteristic compounds of Zhenjiang aromatic vinegars. Copyright © 2012 The Institute of Brewing & Distilling     

镇江香醋陈酿过程中主要呈味物质的分析

采用高效液相色谱及物质口感活性分析镇江香醋陈酿0~8年过程有机酸与核苷酸含量变化,评价镇江香醋的有机酸和核苷酸风味特征。结果表明:在所有醋样中总有机酸含量为95~114 g/L,乳酸和乙酸为主体有机酸（占总有机酸65%以上）。核苷酸总含量为367~532 mg/L;胞嘧啶核苷酸（CMP）、鸟嘌呤核苷酸（GMP）为主要核苷酸（占总核苷酸75%以上）。蛋白质与氨基酸总量在陈酿过程基本稳定,蛋白质含量为20~27 g/L,氨基酸含量为1.5~2.5 g/L。TAV分析表明,酸感呈味中乙酸呈味最强,苹果酸、酒石酸、琥珀酸呈味效果稍次。核苷酸中,仅GMP有明显呈鲜作用,但弱于琥珀酸。主成分分析表明,陈酿时间与香醋品质密切相关。     

实时荧光定量PCR监测镇江香醋醋酸发酵过程中微生物变化

对镇江香醋醋酸发酵阶段醋醅中功能微生物的变化进行定量分析。建立了实时荧光定量PCR方法,对醋酸发酵阶段醋醅中总细菌、总真菌、醋酸菌、乳酸菌和酵母的动态变化进行了定量分析。研究结果表明,发酵起始阶段(1～7天)醋醅中总细菌、醋酸菌和乳酸菌的生物量快速上升,分别于第6、7、4天达到最大值,为4.85×1011,1.14×1010和3.37×1011copies/g干醅。随后各类细菌的生物量逐渐下降,并维持在一定水平。醋醅中总真菌和酵母的生物量在发酵前期变化不大,7天后至发酵结束总真菌的生物量逐渐下降为7.59×104copies/g干醅,而酵母生物量则在发酵8～12天内下降为0。     

优良菌株组合发酵对醋醅中川芎嗪及其前体合成的影响

该研究监测山西老陈醋醋酸发酵和熏醅过程中川芎嗪及其前体物质的变化,并对高产乙偶姻及川芎嗪菌株进行筛选及共培养,优选高产菌株组合,在醋醅模拟培养基中添加优良菌株组合联合底物及前体物质,考察其对乙偶姻和川芎嗪生成的影响。结果表明,丙酮酸、2,3-丁二醇和乙偶姻在醋酸发酵阶段呈现上升趋势,在熏醅阶段,2,3-丁二醇和乙偶姻呈下降趋势,丙酮酸含量变化不大;川芎嗪则主要在熏醅阶段生成,从熏醅第2天的6.78 μg/g急剧上升到熏醅第5天的43.52 μg/g。在醋醅模拟培养基中添加莫海威芽孢杆菌（Bacillus mojavensis）B15+甲基营养型芽孢杆菌（Bacillus methylotrophicus）B6+丙酮酸组合,川芎嗪生成量最高,达到15.76 μg/g,比对照组提高132.79%。     

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     

地理标志产品镇江香醋标准符合性验证分析

目的从理化指标、特征有机酸以及检验方法等对2018年江苏镇江地理标志产品镇江香醋进行标准验证。方法对5个质量等级,共计20批次镇江香醋样品进行抽检,采用GB/T 18623-2011标准对特征性成分进行分析检测,并对检验标准进行验证。结果镇江香醋5个等级产品理化特性均符合标准规定,未检测到致病菌,菌落总数以及大肠菌群均未超过标准规定的限量要求;重金属铅和总砷、真菌毒素黄曲霉毒素B_1以及食品添加剂苯甲酸和山梨酸均符合标准规定技术限量要求;具有镇江香醋标志性特征有机酸(乳酸、乙酸、焦谷氨酸和琥珀酸),且随着等级要求越高,其有机酸含量越高。结论镇江香醋特征性成分完全符合其标准规定要求,镇江香醋标准与国家强制性标准一致,建议可对有机酸的液相检测方法进行优化,尝试离子排斥色谱柱或耐酸C_(18)柱,以满足镇江香醋有机酸液相检测更高的分析要求。     

高效液相色谱法测定镇江香醋醋酸发酵过程中果糖和葡萄糖含量

建立了果糖和葡萄糖的高效液相色谱(HPLC)分析方法,并用于镇江香醋醋酸发酵过程中单糖含量的分析。分析条件:醋醅浸出液用C18固相萃取小柱进行预处理,以Cosmosil Sugar-D作为分离柱,乙腈∶水(78∶22,v/v)为流动相,柱温35℃,流速1.0 mL/min,示差折光检测器进行检测。结果表明,该方法具有良好重复性和准确度,平均加标回收率为96.28%～96.69%。该研究结果为食醋酿造过程中果糖和葡萄糖含量的定量分析提供了一种高效快速的测定方法。     

GC-MS/MS测定镇江香醋中的氨基甲酸乙酯

镇江香醋中的氨基甲酸乙酯(EC)通过乙酸乙酯提取,用N-丙基乙二胺(PSA)分散固相萃取的方法净化,采用气相色谱-串联质谱(GC-MS/MS)法测定。结果表明,氨基甲酸乙酯(EC)的回收率达到96.7%,标准曲线的相关系数R2为0.999 7,最低检出限为1 μg/kg。该方法简单、快速、灵敏,能有效减少杂质干扰。测定二十份不同厂家生产的镇江香醋,均检出氨基甲酸乙酯（EC）。表明镇江香醋在生产发酵过程中,不可避免的会发生EC污染,应采取措施进行控制。     

电子鼻分析镇江香醋发酵和陈酿阶段的气味变化

利用PEN3电子鼻对镇江香醋发酵和陈酿过程中挥发性风味进行分析。结果表明,电子鼻能准确稳定地识别醋醅和陈酿醋气味图谱的变化并对不同发酵时间的醋醅和陈酿醋中的败坏醋进行区分;醋醅和食醋的挥发物组成类似,主要是氮氧化合物类、甲烷类、硫化物类、醇类、有机硫化物类;线性判别分析（LDA）和主成分分析（PCA）都能区分各个样品,但PCA对醋醅的区分优于LDA,载荷分析结果表明,醋醅中甲烷类和氮氧化合物类分别对第一、二主成分方差贡献率最大,醋液中甲烷类和硫化物类分别对第一、二主成分贡献率最大。LDA对败坏醋的区分优于PCA。     

不同封醅时间对镇江香醋品质的影响

本文对镇江香醋不同封醅时间（0、3、7、15、30 d）进行研究,以期更好地控制镇江香醋品质。以不同封醅时间的镇江香醋为研究对象,对总酸、还原糖、不挥发酸、氨基酸态氮、有机酸、氨基酸、挥发性物质等指标含量进行了差异性分析。结果表明:通过封醅工艺可显著（P<0.05）增加醋卤中总酸、不挥发酸和乳酸的含量,封醅15 d以上总酸达到7.30±0.06 g/100 mL,不挥发酸达到2.36±0.03 g/100 mL,乳酸达到1133.57±1.47 mg/100 mL;游离氨基酸含量也随着封醅处理而增加,封醅30 d后增加了101.16 mg/L;成品醋中挥发性风味物质的含量和种类也随着封醅时间的延长而增加,封醅15 d以上的醋醅所酿造的香醋具有较佳的品质。说明采用封醅工艺对镇江香醋风味物质的形成至关重要,通过封醅工艺的镇江香醋品质优于不封醅镇江香醋。     

Effects of exogenous nitric oxide on contents of soluble sugars and related enzyme activities in 'Feicheng' peach fruit

BACKGROUND: Sugar content is one of the main characteristics related to the quality of fruit. Research confirms that nitric oxide (NO) involves a physiological process and prolongs the storage life of fruit. However, little attention has been paid to the effects of NO on sugar metabolism in fruit during storage. In this study, the effect of different concentrations (0, 10, 30 µmol L^?1) of exogenous NO treatment on sugar content and related enzyme activities in ‘Feicheng’ peach fruit was investigated during storage (0–12 days after harvest) at room temperature (25 °C). RESULTS: Results showed that the decrease of firmness and accumulation of sugar and acid:sugar ratio in peach fruit during storage were significantly inhibited by treatment with 10 µmol L^?1 NO. Treatment with 10 µmol L^?1 NO could promote fructose and glucose metabolism during the first 4 days of storage, and increase the content of sucrose and the activities of sorbitol dehydrogenase, sorbitol oxidase and sucrose phosphate synthase in peach fruit during storage. However, acid invertase activity from 8 to 12 days of storage and neutral invertase activity during the first 4 days of storage were inhibited by treatment with 10 µmol L^?1 NO. At the same time, treatment with 10 µmol L^?1 NO inhibited sucrose synthase (SS) activity in decomposition during storage and SS activity in synthesis from 8 to 12 days of storage. CONCLUSION: Treatment with 10 µmol L^?1 NO had a significant impact on content of soluble sugars and related enzyme activities in ‘Feicheng’ peach fruit during storage (0–12 days) at room temperature (25 °C). Copyright © 2011 Society of Chemical Industry     

固态发酵醋生产中铅的关键控制点研究

研究固态发酵食醋生产过程中铅的迁移转化规律,确定食醋生产过程中铅的关键控制点。分别采用Tessier连续提取法和有机滤膜过滤法研究固液相中铅的初级形态及变化。采用原子吸收光谱法测定样品中的铅含量。结果表明液相中的铅主要以可溶态铅为主,醋酸发酵阶段液相中铅含量的增加主要与固相中蛋白结合态铅及草酸铅的迁移转化有关。加工与陈酿过程所使用设备的材质是影响食醋铅含量的主要因素。醋醅对铅有较强的吸附力,因此对液相中的铅含量具有调控作用。食醋生产过程:制醅、发酵、煎煮及陈酿影响液相中可沉淀物含量,而液相可沉淀物具有富集铅的作用。通过本研究表明改进设备材质、采用铅离子吸附剂及分离除去醋液沉淀将是控制食醋中铅含量的有效措施。     

不同加工方式镇江香醋不同季节理化指标

试验比较不同季节生产的传统手工和现代工艺镇江香醋主要理化指标的差异,以更好指导实际生产。以不同季节生产的传统手工和现代工业镇江香醋为对象,对还原糖、总酸、氨基酸态氮、有机酸、游离氨基酸等含量的差异进行研究,得到冬季生产的镇江香醋中还原糖、氨基酸态氮和游离氨基酸含量均为最高,而夏季生产的食醋还原糖、总酸、氨基酸态氮、有机酸和游离氨基酸含量均为最低,说明冬季生产的镇江香醋口感更柔和;4个季节生产的手工醋中还原糖含量均比工业醋低,而手工醋中总酸、氨基酸态氮、有机酸和游离氨基酸含量均比工业醋高,说明手工醋的口感更柔和、醇厚。     

Inhibition of the decrease of volatile esters and terpenes during storage of a white wine and a model wine medium by glutathione and N-acetylcysteine

Glutathione and N‐acetylcysteine, each at 20 mg L^?1, were tested as inhibitors of the decrease of volatile esters and terpenes during storage of Debina white wine. Moreover, the inhibition of the decrease of isoamyl acetate, ethyl hexanoate and linalool in a model wine medium by glutathione and N‐acetylcysteine, each at 0–20 mg L^?1, was also tested. Several volatiles, such as isoamyl acetate, ethyl hexanoate, ethyl octanoate, ethyl decanoate and linalool, decreased during wine storage. Glutathione or N‐acetylcysteine significantly restricted the decrease of these volatiles. In the model medium, each thiol inhibited the decrease of the three volatiles in a dose‐dependent manner. N‐acetylcysteine inhibited the decrease of all three volatiles at 2.5 mg L^?1 while glutathione at 2.5 or 5.0 mg L^?1. The present results indicate that glutathione and N‐acetylcysteine may be taken into account as potent inhibitors of the disappearance of aromatic esters and terpenes in wines.     

Optimisation of lactic acid fermentation of York cabbage for the development of potential probiotic products

In this study, York cabbage was used as the sole substrate for the lactic acid (LA) fermentation with Lactobacillus plantarum. York cabbage was blanched at 95 °C for 12 min to inactivate surface microflora prior to fermentation. To achieve an optimal fermentation condition that would result in higher release of phytochemicals and antioxidant (AO) capacity in the broth, Box‐Behnken design integrating a desirability approach was used. A second‐order polynomial equation was developed, indicating the effect of solute/liquid ratio, agitation rate and fermentation time on desired parameters. The optimised factors were used for fermenting York cabbage to obtain maximum bacterial growth, LA, polyphenols and AO capacity. There was ≈5 log cfu mL^?1 increment in bacterial growth after fermentation, whereas LA production reached up to 4.97 mg mL^?1. Results showed that fermentation retains 95–98% and 90–95% of TPC and AO capacity, respectively. During refrigerated storage (4 °C), the cell numbers, bioactive components and acidity were maintained till 15‐day storage.     

保宁醋醋曲中产多糖乳酸菌的筛选及其代谢产物分析

保宁醋因其复杂微生物体系和酿造工艺而形成独特风味,其风味物质和营养成分主要由各种功能性微生物发酵生成,其中乳酸菌是非常关键的一类菌株,其对保宁醋的风味和营养物质的形成具有重要作用。本研究以稀释涂布平板法从保宁醋醋曲中分离得到12株乳酸菌,通过定性试验、产酸率、耐酸试验和胞外多糖测定等实验,得到一株产多糖量为191.98 mg/L、产酸率为1.62%的乳酸菌L7,鉴定为发酵乳酸杆菌(Lactobacillus femertum),其发酵液中3-羟基-2-丁酮和乙酸相对含量较高,分别为36.22%、45.76%,HPLC检测到乳酸、乙酸,含量分别为66.56 mg/100 mL和104.08 mg/100 mL,这表明L7将有利于提高食醋酸度以及川芎嗪含量。产多糖乳酸菌在食醋发酵过程中的具有重要作用,而此次所得乳酸杆菌对保宁醋生产和发酵工艺改良具有指导意义。     

葡萄果醋饮料的工艺研究

以玉米、麸皮为主要原料,经酒精发酵后加入葡萄皮渣进行醋酸发酵而制得葡萄果醋,再经调配,制成风味独特的葡萄果醋饮料。通过单因素实验、正交试验,优化了工艺条件和配方。结果表明,以玉米、麸皮汁为发酵基料,基料的糖含量为16°Brix,pH 3.5,酵母接种量为10%,发酵温度30℃,经酒精发酵6 d,酒精体积分数达到6%～7%时进入醋酸发酵,糖含量4°Bx,醋酸菌接种量10%,发酵温度30℃,醋酸发酵6 d。风味配方为:葡萄果醋10%,柠檬酸0.1%,蔗糖9%。