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。     

葡萄酒中生物胺的研究进展

生物胺是一类低分子量的含氮化合物,普遍存在于葡萄酒和其他发酵食品中。葡萄酒中的生物胺主要是由相关微生物对氨基酸脱羧而产生,并且与诸多因素相关。利用多重PCR和实时定量PCR技术可以实现对葡萄酒中生物胺产生菌的快速定量检测。高效液相色谱法（HPLC）、薄层色谱法（TLC）、离子色谱法（IC）、毛细管电泳法（CE）、气相色谱法（GC）可以定量测定葡萄酒中的多种生物胺,其中,以高效液相色谱法最为常用。通过接种优良的酵母菌和无氨基酸脱酶活性的乳酸菌,及时终止苹-乳发酵（MLF）,并避免产生污染菌,可实现对葡萄酒中生物胺的控制。     

不同接种方式下小片球菌C30对‘赤霞珠’葡萄酒品质的影响

以甘肃河西走廊葡萄酒产区小片球菌C30为苹果酸乳酸发酵(malolactic fermentation,MLF)启动菌,比较以不同接种方式进行MLF后对葡萄酒品质的影响,探讨小片球菌C30用于MLF的可行性及其优势。将小片球菌C30在酒精发酵(alcohol fermentation,AF)的早、中、后期进行接种,以商品乳酸菌为对照进行酿造实验。在研究小片球菌C30的MLF动力学与酿造葡萄酒安全性的基础上,分析其对葡萄酒基本理化指标、香气成分和感官品质等的影响。结果表明,MLF后期接种小片球菌C30,完成发酵时间最短(27 d),酒样中多糖、生物胺等物质的含量和残糖、酒精度等理化指标符合国家标准,酒样的香气物质总含量显著高于商品菌株。小片球菌C30可完成MLF,且具有安全性,在AF后期接种进行MLF,性能优良,对提升赤霞珠葡萄酒品质有积极影响。     

生物传感器测定宝石-卡本内特干红葡萄酒苹果酸-乳酸发酵过程中L-乳酸含量

苹果酸是葡萄酒中重要的有机酸之一,其生理代谢比较活跃,易被微生物分解利用,在葡萄酒酿造工艺中起关键作用。葡萄酒的苹果酸-乳酸发酵（以下简称MLF）是乳酸菌以L-苹果酸为底物,在苹果酸-乳酸酶催化下转变成L-乳酸和CO2的过程。酸味尖刻的二元酸L-苹果酸向酸味柔和的一元酸L-乳酸的转化,使葡萄酒总酸下降,酸涩感降低,使酒体口感酸味柔和协调,果香突出,提高葡萄酒的品质。如果对MLF控制不当,就有可能给葡萄酒带来刺激的酸乳味、苦昧等,还可产生某些挥发性的酚类或生物胺（如组胺、腐胺、尸胺等）,     

一株降解生物胺乳酸菌用于葡萄酒苹果酸乳酸发酵的特性研究

葡萄酒等发酵食品中微生物通过脱羧作用形成生物胺，高浓度的生物胺对人体有害。该研究以课题组前期分离得到的一株可降解生物胺的Lactobacillus plantarum NXU-Q12为试验菌株，将其应用于葡萄酒酿造。采用HPLC法测定该菌株的生物胺降解率；采用顶空固相微萃取-气相色谱-质谱联用(head space-solid phase microextraction-gas chromatography-mass spectrometer, HS-SPME-GC-MS)和电子鼻测定经该菌株苹果酸-乳酸发酵(malolactic fermentation, MLF)前后的葡萄酒中挥发性化合物，以此评价该菌株对葡萄酒品质的影响，为葡萄酒酿造过程中生物胺的控制提供解决途径。结果表明，L.plantarum NXU-Q12在发酵葡萄酒的过程中具有降解苹果酸而不降解酒石酸、柠檬酸和甘油的特点，且MLF后酒样(赤霞珠葡萄酒、马瑟兰葡萄酒)中的酒精度、总酸、还原糖、挥发酸、甘油含量以及pH值均符合葡萄酒国家标准。此外，赤霞珠、马瑟兰酒样中L-苹果酸含量在MLF 32 d后均降至0.1 g/L以下...     

葡萄酒中生物胺含量的影响因素研究

研究了葡萄酒生产过程中影响生物胺产生量的主要因素,结果表明,葡萄酒酒精发酵和苹果酸．乳酸发酵过程均有牛物胺的产生,影响葡萄酒中生物胺产生的主要因素是乳酸菌中氨基酸脱羧酶的活性,本研究添加了氨基酸脱羧酶辅酶的样品与对照相比,生物胺含量提高了90．6％。酵母菌和乳酸菌种类、酒精发酵温度,生物胺前体物质也是影响生物胺含量的主要因素,发酵温度越高、生物胺产生量越;在一定范围内,氨基酸的浓度越大产生物胺越多。但酵母菌和乳酸菌的添加量对生物胺浓度的影响较小。     

微生物降酸技术在葡萄酒酿造中的应用

较全面地介绍了葡萄酒微生物降酸技术的主要内容.包括乳酸菌进行的苹果酸-乳酸发酵(MLF),裂殖酵母进行的苹果酸-酒精发酵(MAF)以及转基因酵母进行的MLF等降酸方法.本文对MLF和MAF生化反应机理、影响MLF和MAF的环境因素以及现代发酵工程技术(固定化细胞、酶技术、膜生物反应器)在葡萄酒生物降酸工艺中的应用也进行了回顾.     

两种乳酸菌在干红葡萄酒中MLF的比较

将两种乳酸菌MBR和VINIFLORA分别接种于赤霞珠干红葡萄酒中进行苹果酸-乳酸发酵(MLF).结果表明,在试验条件下,乳酸菌MBR的降酸幅度和降酸速率均高于乳酸菌VINIFLORA;MBR产生的挥发酸低于乳酸菌VINIFLORA;pH值变化在正常范围内;影响MLF的主要因素是SO2的含量和pH值的高低.说明在进行MLF时,乳酸菌MBR的发酵特性优于乳酸菌VINIFLORA.     

酒类酒球菌对葡萄酒中相关物质代谢的影响

酒类酒球菌是触发葡萄酒苹果酸-乳酸发酵(malolactic fermentation,MLF)的主要乳酸菌,其代谢产物如乳酸、乙酸、双乙酰、乙偶姻、生物胺等对葡萄酒的感官品质有重要作用。本文对酒类酒球菌在葡萄酒MLF过程的代谢情况进行总结,阐述糖、有机酸、含氮物质和酚类化合物的代谢情况,以及在不同影响因素下酒类酒球菌生长和代谢途径的改变状况,已经证明pH值、SO2、乙醇体积分数、葡萄糖/果糖比等都是影响代谢的因素。随着研究的不断深入,酚类化合物在代谢中的作用也受到越来越多的关注。酚类化合物一方面通过刺激或者抑制酒类酒球菌的生长而间接影响MLF的进行;另一方面,不同的酚类物质也会影响细菌对其他酚类的代谢,从而影响葡萄酒的口感和质量。通过全面综合了解酒类酒球菌在葡萄酒中的代谢情况有助于更好地调控MLF过程。     

葡萄酒酿造过程中不同工艺参数对生物胺含量的影响

为揭示葡萄酒酿造过程中生物胺产生积累的关键控制因素,有效降低葡萄酒中生物胺的含量。本试验选取不同酿酒参数条件进行酿酒试验,利用高效液相色谱进行检测,分析比较不同工艺参数及相关因素对葡萄酒中生物胺含量的影响。结果表明,氨基酸脱羧酶的活性是影响生物胺含量的重要因素,酵母类型及乳酸菌类型对于生物胺含量的影响高于其接种量对生物胺含量的影响,且发酵完成后,贮存的温度越高,其生物胺含量也越高。     

Evaluating the Influence of Malolactic Fermentation Inoculation Practices and Ageing on Lees on Biogenic Amine Production in Wine

Biogenic amines are compounds, produced primarily by lactic acid bacteria (LAB) that negatively affect the wholesomeness of wine. Standard winemaking practices can greatly influence the levels of biogenic amines in wine. The aim of this study was to determine the relative contribution of different malolactic fermentation (MLF) practices and ageing of wines on fermentation lees to the final levels of biogenic amines. Wines were made on small scale over two harvest seasons with two red grape cultivars. Treatments included spontaneous MLF, co-inoculated MLF, MLF inoculated after alcoholic fermentation (conventional inoculation) and 4 months of ageing in the presence and absence of fermentation lees of all MLF treatments. Biogenic amine concentrations were measured by high-performance liquid chromatography at key winemaking stages and statistically analysed for the effects of MLF treatment and winemaking stage. Results indicate that the presence of indigenous LAB increased the risk of biogenic amine formation. Inoculation proved to reduce biogenic amine production over time compared to spontaneous MLF and co-inoculation even more than conventional inoculation. The presence of yeast lees during ageing generally led to higher final concentrations of biogenic amines in wines than the absence of lees. This study confirms other works that conclude that spontaneous MLF and uncontrolled ageing on yeast lees are generally unpredictable and pose a risk of biogenic amine contamination in finished wines.     

发酵过程果酒成分对苹果酸乳酸发酵的影响

苹果酸-乳酸发酵（MLF）是果酒（葡萄酒、苹果酒）酿造中非常重要的二次发酵过程。本研究采用人工模拟果酒，研究果酒成分对MLF的影响。结果表明，葡萄糖抑制乳酸菌的MLF，果糖却有促进作用，而且果糖可解除葡萄糖对酿酒酒球菌MLF的抑制作用，果酒中葡萄糖和果糖的浓度及其比例是预测和控制MLF的重要参数；高浓度乙醇、低pH抑制乳酸菌的MLF，其抑制作用的大小与使用的菌种有关；酒中酚类物质（没食子酸和阿魏酸）对酿酒酒球菌的MLF几乎无任何显著影响；酒精发酵完成后果酒中营养的缺乏会抑制乳酸菌的MLF。     

Malolactic fermentation of Tempranillo wine: contribution of the lactic acid bacteria inoculation to sensory quality and chemical composition

Malolactic fermentation (MLF) of Tempranillo Rioja wines (Spain) inoculated with two lactic acid bacteria (LAB) strains were studied and compared with spontaneous MLF. Inoculation with selected Oenococcus oeni lyophila shortened MLF duration up to 19 days and lead to wines with more fresh and fruity characters, especially when implantation was 100%. We demonstrated modifications in the concentration of volatile and nitrogenous compounds and a good correlation between analytical and sensory attributes was also noted. In addition, the low initial amino acid concentration and the consumption of these compounds by the inoculated yeast strain during alcoholic fermentation resulted in wines with very low biogenic amines levels (under 3.75 mg L^?1) after MLF and 3 month storage period in all cases. The results showed the significance of choose the most suitable starter to elaborate quality wines and suggest the control of amino acid content in must and wine to prevent the formation of biogenic amines.     

Fast identification of wine related lactic acid bacteria by multiplex PCR

The microflora of must and wine consists of yeasts, acetic acid bacteria and lactic acid bacteria (LAB). The latter group plays an important role for wine quality. The malolactic fermentation carried out by LAB leads to deacidification and stabilisation of wines. Nevertheless, LAB are often associated with wine spoilage. They are mainly responsible for the formation of biogenic amines. Furthermore, some strains produce exopolysaccharide slimes, acetic acid, diacetyl and other off-flavours. In this context a better monitoring of the vinification process is crucial to improve wine quality. Moreover, a lot of biodiversity studies would also profit from a fast and reliable identification method.     

Screening of biogenic amine production by lactic acid bacteria isolated from grape must and wine

The potential to produce the biogenic amines tyramine, histamine and putrescine, was investigated for lactic acid bacteria (LAB) of various origin, including commercial malolactic starter cultures, type strains and 78 strains isolated from Spanish grape must and wine. The presence of biogenic amines in a decarboxylase synthetic broth was determined by reverse-phase high performance liquid chromatography (RP-HPLC). Tyramine was the main amine formed by the LAB strains investigated. Leuconostoc strains were the most intensive tyramine formers. No potential to form biogenic amines was observed in Oenococcus oeni strains. Two strains of Latobacillus buchneri were associated with putrescine formation. None of the lactic acid bacteria produced histamine. According to these in vitro results, the commercial starter bacteria analyzed did not produce histamine, tyramine and putrescine.     

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.     

黄酒前酵中生物胺生成规律的研究

本研究对黄酒前酵工序的生物胺生成规律及影响因素进行了分析探讨。采用反相高效液相色谱技术,改进了生物胺定量检测法,该检测体系准确可靠,样品峰型对称分离度好且缩短了检测时间。分析了前酵工序中主要微生物,氨基酸,发酵醪酸度、糖度、酒精度、p H对生物胺生成的影响。发现前酵工序对成品酒生物胺的影响度为77.67%,第一次开耙阶段生物胺总量增幅最大,达到7.63 mg/L。研究发现主要酿造微生物中乳酸菌总数与生物胺总量成正相关,乳酸菌在搭窝期间生长速率最大,为7.13×106CFU/(m L·h)。在前酵工序中生物胺总量与发酵醪酸度、酒精度、p H呈显著正相关,与糖度呈负相关;前酵过程中主要生物胺与前体氨基酸也呈明显正相关。本研究分析了黄酒前酵中生物胺生成规律,有助于建立降低黄酒生物胺含量的更安全、科学的工艺。     

Metabolism of SO₂ binding compounds by Oenococcus oeni during and after malolactic fermentation in white wine

Sulfur dioxide SO? is the key additive for the preservation of wines. Carbonyl and keto compounds in wine can bind to SO? and decrease its efficacy, resulting in higher total SO? requirements. Increased consumer demand for low sulfite and organic wines pose production challenges if SO? binders have not been properly managed during vinification. Malolactic fermentation (MLF) has been known to reduce bound SO? levels but detailed time course studies are not available. In this work, the kinetics of major SO? binding compounds and malic acid were followed during MLF in wine with 12 commercially available strains of Oenococcus oeni. Pyruvic acid, acetaldehyde and α-ketoglutaric acid were degraded to various degrees by O. oeni, but galacturonic acid was not. At the time of malic acid depletion, percent degradation of pyruvate, α-ketoglutaric acid and acetaldehyde was 49%, 14% and 30%, respectively. During MLF, the decrease in average bound SO? levels, as calculated from carbonyl metabolism, was 22%. The largest reduction in wine carbonyl content occurred in the week after completion of MLF and was 53% (107 mg/L to 34 mg/L) calculated as bound SO?. Prolonged activity of bacteria in the wines (up to 3 weeks post malic acid depletion) resulted only in reduced additional reductions in bound SO? levels. The results suggest that microbiological wine stabilization one week after malic acid depletion is an effective strategy for maximum removal of SO? binders while reducing the risk of possible post-ML spoilage by O. oeni leading to the production acetic acid and biogenic amines.