产淀粉酶非酿酒酵母菌的筛选及产酶特性研究

通过对非酿酒酵母菌的筛选鉴定,共得到11株产淀粉酶菌株,酶学性质研究表明,该11株酵母菌在pH值为7~9时表现了较高的酶活力,在pH值为5~6时保持稳定;最适反应温度为40℃,在30~50℃时保持较高酶活力;金属离子依赖性发现,Fe2+、Mg2+对酶有不同程度的激活作用,Ca2+、Ba2+、Cu2+对酶有抑制作用;十二烷基硫酸钠(SDS)、乙二胺四乙酸(EDTA)对酶均有抑制作用,EDTA抑制酶活约为50%,淀粉酶对二价金属离子具有一定的依赖作用。     

桑葚酒用非酿酒酵母的筛选及特性研究

为筛选出适合桑葚酒发酵的非酿酒酵母,改善桑葚酒的口感及香味,以桑葚及桑园土壤样品中筛选得到的72株酵母菌为出发菌株,经杜氏管发酵、酒精发酵与感官品评进行初筛。将初筛得到的酵母分别与酿酒酵母（Saccharomyces cerevisiae）BO213按1∶1的比例接种到桑葚汁中发酵,通过感官品评与理化分析优选出一株非酿酒酵母JM-7,经鉴定该酵母为异常威克汉逊酵母（Wickerhamomyces anmalus）。其与菌株BO213混酿的桑葚酒感官评分为90.2分,总酯含量为3.21 g/L,还原糖含量为3.9 g/L,酒精度、总酸、挥发酸含量分别为11.7%vol、7.97 g/L、0.51 g/L。研究菌株JM-7生长曲线及耐受性发现,该酵母生长速率快,5 h即进入对数期,36 h达到稳定期,最高菌体浓度为4.5×108 CFU/mL,对温度、pH、SO2都有较好的耐受性,在16～36 ℃、pH 2～10、SO2质量浓度0～100 mg/L均能正常生长,但对酒精的耐受性较差,在酒精度为8%vol时生长受到抑制,在12%vol时不能生长。     

酿酒葡萄表皮产酶非酿酒酵母的筛选及其产酶特性研究

非酿酒酵母是葡萄酒发酵过程中主要微生物之一,对葡萄酒品质具有重要影响。该研究从新疆昌吉的酿酒葡萄表皮分离产酶非酿酒酵母,基于26S rDNA序列对菌株进行分子生物学鉴定,并研究其产酶特性。结果表明,从酿酒葡萄表皮分离到酵母菌株62株,其中毕菌株CZ4（赤克鲁维酵母（Pichia kluyveri））产β-葡萄糖苷酶活性最高为214.57 U/mL,菌株KS5（葡萄有孢汉逊酵母（Hanseniaspora uvarum））产蛋白酶活性最高为178.56 U/mL,菌株KS3（仙人掌有孢汉逊酵母（Hanseniaspora opuntiae））产脂肪酶活性最高为184.68 U/mL。随着培养时间与SO2、糖以及酒精度的增加,3株高产酶酵母对糖、酒精和SO2耐受性整体呈现先增加后减少的趋势。     

非酿酒酵母与酿酒酵母混合发酵对葡萄酒香气的影响

为研究自选非酿酒酵母对葡萄酒香气物质的影响,选取毕赤克鲁维酵母（Pichia kluyveri）HSX-5、长孢洛德酵母（Lodderomyces elongisporus）MNS-6和戴尔有孢圆酵母（Torulaspora debrueckii）YQX-8与酿酒酵母（Saccharomyces cerevisiae）AWRI 796混合发酵赤霞珠葡萄酒,采用顶空固相萃取-气相色谱质谱联用（HS-SPME-GC-MS）技术检测其挥发性香气物质,并对结果进行主成分分析（PCA）。结果表明,相较于酿酒酵母单菌发酵酒样,菌株HSX-5和YQX-8混合发酵酒样中己酸乙酯提升了2倍,菌株HSX-5、YQX-8、MNS-6混合发酵酒样中的辛酸乙酯分别提升76.6%、35.5%、5.5%,乙酸乙酯含量增加均为10%,混合发酵增强了葡萄酒花果类香气及醇香;同时混合发酵各组均产生了新物质,增加了葡萄酒风味体系的丰度。PCA结果表明,菌株HSX-5和YQX-8混合发酵的发酵酒样主要贡献香气种类较多且气味活度值（OAV）较高,对葡萄酒香气的影响显著。     

混菌顺序发酵对赤霞珠干红葡萄酒品质的影响

为研究不同非酿酒酵母与酿酒酵母混合发酵对葡萄酒品质的影响,选用2种商业非酿酒酵母（编号为CT10、PL09）分别与3种商业酿酒酵母（编号为MST、CEC01、F33）以1∶1混合顺序发酵,得到赤霞珠葡萄干红葡萄酒样（编号为CM、CC、CF、PM、PC、PF）,并分析各酒样的理化指标、颜色参数、酚类物质含量及感官品质。结果表明,与酿酒酵母单菌发酵相比,混菌发酵将发酵期延缓了2～3 d,总酸、干浸出物含量分别增加3.20%～7.75%和0.20%～3.47%,且提升总色素、聚合色素的稳定性,并提高酚类物质含量。其中酒样CM、PF中单宁、总酚、总黄酮含量分别提升2.30%、17.74%、44.68%,50.96%、19.02%、15.34%;酒样CM总花色苷含量提升22.90%,而酒样PF降低31.25%;酒样CM、PF香气复杂,口感圆润饱满,感官评分均为“优秀”,分别比对照提高35.81%、22.23%。综上,添加非酿酒酵母混菌发酵有助于提升赤霞珠干红葡萄酒的品质。     

葡萄酒酵母选育技术研究进展

文中主要从酿酒酵母及非酿酒酵母的选育展开论述,概括介绍了目前葡萄酒酵母的选育技术及成果,同时根据文献对国内和国外的研究现状作了分析,并对今后的研究趋势做了推测.     

Screening of non-Saccharomyces wine yeasts for the production of beta-D-xylosidase activity

Fifty-four yeast strains belonging to the genera Candida, Dekkera, Hanseniaspora, Metschnikowia, Pichia, Rhodotorula, Schizosaccharomyces and Zygosaccharomyces, mainly isolated from grapes and wines, were screened for the production of beta-D-xylosidase activity. Beta-D-xylosidase activity was only detected in eight yeast strains belonging to the genera Hanseniaspora (H. osmophila and H. uvarum) and Pichia (P. anomala). Beta-D-xylosidase preparations active against p-nitrophenyl-beta-D-xyloside were characterised with respect to their optimal pH and temperature conditions. H. uvarum 11105 and 11107 and P. anomala 10320 beta-D-xylosidase preparations were active at pH and temperature ranges and at concentrations of glucose and ethanol usually found during winemaking processes.     

非酿酒酵母与酿酒酵母混合发酵柿子酒特性的研究

基于非酿酒酵母代谢产物可提升酒的香气和风味物质,将两种非酿酒酵母柠檬形克勒克酵母（Klockera apiculata）和东方伊萨酵母（Issatchenkia orientalis）用于柿子酒发酵,研究两种非酿酒酵母对酒精、二氧化硫、单宁的耐受性和产β-葡萄糖苷酶的能力;对它们分别与酿酒酵母（Saccharomyces cerevisiae）混合发酵柿子酒的特性进行研究。结果表明,东方伊萨酵母在各方面耐受性均优于柠檬形克勒克酵母,但产β-葡萄糖苷酶的能力却较弱;两种非酿酒酵母单菌发酵性能都较弱,东方伊萨酵母单菌发酵结束酒精度为4.51%vol,而柠檬形克勒克酵母单菌发酵酒精度只有3.05%vol;两种非酿酒酵母与酿酒酵母混菌发酵7 d后,酒精度均达到10%vol以上。     

草莓果酒酵母菌的筛选、鉴定及耐受性研究

从自然发酵的草莓果酒中分离得到野生酵母菌,根据其菌落形态特征、生理生化实验及酒精、二氧化硫、葡萄糖、pH的耐受性实验,筛选出性能最优2株菌株（RL4和RL6）进行分子生物学鉴定。结果表明,初步鉴定菌株RL4和RL6分别为Hanseniaspora thailandica和葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）。该研究为草莓果酒研制提供了优良酵母菌。     

GC-MS对非酿酒酵母菌发酵赤霞珠葡萄酒香气成分的检测

以新疆石河子地区葡萄自然发酵过程中分离鉴定的7株非酿酒酵母菌为主要发酵菌种,以工业菌株为对照,研究了混合发酵赤霞珠葡萄酒香气成分的气相色谱-质谱联用仪分析结果。结果表明,共检测到香气成分16种,其中酯类物质7种,醇类6种,其他香气物质3种。以Y10（Hanseniaspora sp.）孢汉逊酵母属发酵的7#赤霞珠葡萄酒的主要优势特征香气为：苯乙醇、1,3-丁二醇、2-甲基-丙醇、1-己醇、辛酸乙酯、丁酸乙酯、己酸乙酯、琥珀酸二乙酯、乙酸己酯,且相对含量高于其他菌株和对照工业菌株。     

八种商业酵母制备蓝莓苹果复合果酒的品质对比

为筛选出适合制备蓝莓苹果复合果酒的酵母,以蓝莓汁和苹果汁为原料,选用8种国内外商业酵母,对比各种酵母发酵过程 中总酸、总糖及还原糖含量的变化规律,分析蓝莓苹果复合果酒的感官指标和理化指标,并采用顶空固相微萃取-气质联用（HS-SPME- GC-MS）法检测其香气组分。 结果表明,酵母JK10是制备蓝莓苹果复合果酒最佳酿酒酵母,其成品各理化指标均符合GB/T 32783—2016《蓝莓酒》相关要求,感官评分最高87.0分,酒精度为14.9%vol,共检测出58种香气物质,为其产业化开发提供理论基础。     

Optimisation of preservatives for dealcoholised red wine using a survival model for spoilage yeasts

To optimise the application of preservatives in a new commercial dealcoholised red wine (DRW), a survival model of Rhodotorula mucilaginosa and Saccharomyces cerevisiae was developed. A factorial design included three variables, each at three levels: potassium sorbate, sodium metabisulphite and dimethyl dicarbonate (0, 100 and 200 mg L^?1). DRW samples were inoculated with a cocktail of yeasts and counts were performed after 1, 2, 3, 4, 6 and 8 weeks of incubation. Logistic regression analysis was used to create survival models. Potassium sorbate and sodium metabisulphite did not show any fungicide effect alone, while the model predicts that dimethyl dicarbonate at 185 mg L^?1 has fungicide activity. The interaction of potassium sorbate and dimethyl dicarbonate was statistically significant, so both could act as synergic preservatives. The logistic model obtained was validated with a 100% of accuracy for the conditions tested. Combining use of the studied preservatives reduced the individual dose for preservation of DRW.     

香格里拉产区特色非酿酒酵母及自然发酵葡萄酒香气成分的研究

以香格里拉产区澜沧江流域4个不同海拔葡萄园（布村、溜筒江、斯农和阿东）的酿酒葡萄赤霞珠为试材,对各地葡萄酒特色非酿酒酵母进行分离鉴定,并采用气质联用（GC-MS）法对各地自然发酵葡萄酒的香气成分进行分析。结果表明：各地赤霞珠自然发酵葡萄酒中共分离鉴定到非酿酒酵母9个属10个种,其中,葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）在各地均有分离到;黏红酵母（Rhodotorula glutinis）在布村、溜筒江和阿东均有分离到;出芽短梗霉（Aureobasidium pullulans）、汉逊德巴利酵母（Debaryomyces hansenii）和美极梅奇酵母（Metschnikowia pulcherrima）为斯农独有;而核果梅奇酵母（Metschnikowia fructicola）、季也蒙毕赤酵母（Meyerozyma guilliermondii）和克鲁维毕赤酵母（Pichia kluyveri）为阿东独有;浅黄隐球酵母（Cryptococcus flavescens）为溜筒江独有。这些非酿酒酵母使得各地自然发酵葡萄酒的香气各具特色,其中以醇类、酯类和芳香族类香气种类与含量的差异最为突出。     

高产酸本土非酿酒酵母菌株的筛选及发酵性能研究

以25株本土非酿酒酵母菌为研究对象,采用酵母浸出粉胨葡萄糖（YPD）10培养基及Triple M改良模拟汁初筛,并进行耐受性（乙醇、SO2、糖及pH）测定及葡萄汁发酵,筛选能够有效增加葡萄酒酸度的优良本土非酿酒酵母菌。结果表明,非酿酒酵母菌株LT1及HU4产酸性能较好,具有较好的乙醇、SO2、糖和pH耐受性,其中菌株LT1能耐受乙醇体积分数12%、糖400 g/L及pH 2.75,菌株HU4能耐受乙醇体积分数6%vol、糖250 g/L及pH 2.75。菌株LT1和HU4在葡萄汁中启酵时间较短,发酵旺盛期CO2质量损失速率均＞0.8 g/（L·h）,乳酸产量分别为0.93 g/L、1.14 g/L,乙酸产量分别为0.38 g/L、0.42 g/L,具备酿造增酸葡萄酒的潜力。     

Studies on acetate ester production by non-saccharomyces wine yeasts.

A double coupling bioreactor system was used to fast screen yeast strains for the production of acetate esters. Eleven yeast strains were used belonging to the genera Candida, Hanseniaspora, Metschnikowia, Pichia, Schizosaccharomyces and Zygosacharomyces, mainly isolated from grapes and wine, and two wine Saccharomyces cerevisiae strains. The acetate ester forming activities of yeast strains belonging to the genera Hanseniaspora (Hanseniaspora guilliermondii and H. uvarum) and Pichia (Pichia anomala) showed different substrate specificities and were able to produce ethyl acetate, geranyl acetate, isoamyl acetate and 2-phenylethyl acetate. The influence of aeration culture conditions on the formation of acetate esters by non-Saccharomyces wine yeast and S. cerevisiae was examined by growing the yeasts on synthetic microbiological medium. S. cerevisiae produced low levels of acetate esters when the cells were cultured under highly aeration conditions, while, under the same conditions, H. guilliermondii 11104 and P. anomala 10590 were found to be strong producers of 2-phenylethyl acetate and isoamyl acetate, respectively.     

Selected non-Saccharomyces wine yeasts in controlled multistarter fermentations with Saccharomyces cerevisiae

Non-Saccharomyces yeasts are metabolically active during spontaneous and inoculated must fermentations, and by producing a plethora of by-products, they can contribute to the definition of the wine aroma. Thus, use of Saccharomyces and non-Saccharomyces yeasts as mixed starter cultures for inoculation of wine fermentations is of increasing interest for quality enhancement and improved complexity of wines. We initially characterized 34 non-Saccharomyces yeasts of the genera Candida, Lachancea (Kluyveromyces), Metschnikowia and Torulaspora, and evaluated their enological potential. This confirmed that non-Saccharomyces yeasts from wine-related environments represent a rich sink of unexplored biodiversity for the winemaking industry. From these, we selected four non-Saccharomyces yeasts to combine with starter cultures of Saccharomyces cerevisiae in mixed fermentation trials. The kinetics of growth and fermentation, and the analytical profiles of the wines produced indicate that these non-Saccharomyces strains can be used with S. cerevisiae starter cultures to increase polysaccharide, glycerol and volatile compound production, to reduce volatile acidity, and to increase or reduce the total acidity of the final wines, depending on yeast species and inoculum ratio used. The overall effects of the non-Saccharomyces yeasts on fermentation and wine quality were strictly dependent on the Saccharomyces/non-Saccharomyces inoculum ratio that mimicked the differences of fermentation conditions (natural or simultaneous inoculated fermentation).     

机械化生产黄酒酵母菌生物学特性的研究

机械化生产黄酒（半甜型、半干型、干型）的发酵过程是糖化和以多品种、高密度的酵母与乳酸杆菌（细菌）协同作用的混合发酵过程（即：边糖化、边酵母发酵、边乳酸杆菌或细菌发酵同时进行的三边发酵）。基于“三边发酵”理论中对机械化生产黄酒对酵母菌生物学特性的要求,研究了黄酒酵母菌的生物学特性,筛选出了适宜黄酒机械化生产用的优良酵母菌株。     

非酿酒酵母在发酵过程中的代谢产物对葡萄酒质量的影响

非酿酒酵母广泛存在于葡萄园及酿酒环境中,虽然其在发酵过程中能产生各种异味物质影响酒的品质,但某些非酿酒酵母的代谢产物却能增强葡萄酒的香气、风味和复杂性。该文综述了非酿酒酵母在发酵过程中的代谢产物及其对葡萄酒质量的影响,以期对我国筛选非酿酒酵母用于生产具地区特色的葡萄酒提供参考。