葡萄酒中酯类化合物研究进展

酯类是葡萄酒中重要的芳香物质之一,赋予葡萄酒类似新鲜水果的香气。葡萄酒中酯类化合物的种类与含量对葡萄酒最终香气的形成十分重要。目前,对葡萄酒中酯类的研究主要集中于酯类对香气的贡献、合成与水解机制以及酶和基因调控等方面。本文就葡萄酒中酯类的呈香特征、形成途径和酶及编码基因对酯类调控三方面进行了综述,并指出酯类化合物未来研究的方向,以期为研究者提供理论依据。     

脂肪酶在白酒酯类化合物合成中的作用研究进展

酯类化合物是白酒中最重要的一类风味物质,其含量影响白酒的品质和香型。鉴于酯类化合物对白酒风格具有重要影响,研究白酒酿造过程中酯类化合物形成的影响因素具有重要意义。经系统研究并分析白酒酿造过程可知,白酒中酯类化合物主要来源于酸-醇的酶促反应,并且确定了酯化酶中的脂肪酶对白酒酯类化合物合成具有重要的酶促催化作用。为此,本文分析了白酒酿造过程中酯类化合物形成的途径,进而简单介绍脂肪酶及其催化酯类化合物合成机制,论述脂肪酶在白酒酯类化合物形成的研究现状,并探索脂肪酶在白酒酯类化合物形成中的研究前景。     

中国传统白酒风味酯合成研究进展

传统发酵食品是我国食品工业中的重要分支,而白酒则是传统发酵食品的重要组成部分。开放式的固态生态发酵方式赋予了白酒独特的风味属性,其中酯类化合物是白酒中最重要的一类香味成分,其组成和含量与酒体感官、品质密切相关。研究表明微生物合成代谢途径和酶促反应是白酒酿造过程中酯类物质积累的主要生物途径。文章系统总结了白酒酿造过程中酯类化合物的种类,在此基础上,概述了微生物和酯合成酶在风味酯合成方面的研究现状,并对未来风味酯合成的研究方向进行了展望。     

酿酒酵母醇酰基转移酶的研究进展

酵母发酵产生酯类化合物是白酒发酵过程中的重要产香成分之一。在白酒多年的发展过程中,其香味物质的成分一直不能为人们标准化,不能实现更好的机械化智能化的生产,因此研究酵母产酯途径已成为人们现在热议的话题。该文综述了酿酒酵母酰基酯合成途径中的关键酶的酶学性质、晶体结构与分子催化机制,并展望了其在提升白酒品质和推进白酒的标准化进程的潜力与价值。     

基于宏基因组技术分析自然发酵羊肉香肠中微生物多样性及挥发性风味功能基因

利用宏基因组技术测定自然发酵过程中羊肉香肠的微生物群落演替,并运用直系同源蛋白分组比对、京都基因与基因组百科全书和碳水化合物活性酶数据库对挥发性风味物质代谢途径、参与代谢的微生物和酶进行注释与分析。结果发现,羊肉香肠样品中流感嗜血杆菌、金黄色葡萄球菌、酒类酒球菌等是发酵过程的优势菌种,样品发酵至14 d腐生葡萄球菌与马胃葡萄球菌相对丰度达到最大值（16.52%与10.53%）。在发酵0、5、14 d和26 d样品中,发酵5 d样品注释到的基因数最多,发酵过程中碳水化合物代谢和氨基酸代谢是注释最多的代谢途径,糖苷水解酶与糖基转移酶是数量最多的碳水化合物酶。有167个基因参与氨基酸代谢所需酶的编码,碳水化合物代谢途径所需酶由217个基因编码,脂肪酸代谢途径中共有92个基因参与了相关酶的编码,参与3条代谢途径的酶注释到的微生物主要是葡萄球菌属、明串珠菌属、假单胞菌属、嗜冷杆菌属、弧菌属等,发酵14 d样品中3条代谢途径的大部分酶丰度达到最大值。研究结果对于解析羊肉香肠中微生物的动态变化以及挥发性风味物质的形成机理具有重要的参考价值。     

油酸诱导对灵芝三萜生物合成途径中关键酶编码基因的影响

通过实时荧光定量PCR技术考察油酸诱导对灵芝三萜生物合成途径中关键酶基因转录水平的影响。主要选择SQS、CYP51和14α-LDM3个关键酶基因做试验。在不同浓度油酸、不同诱导时间和最佳诱导条件下,3个基因响应油酸诱导的变化情况各不相同。结果表明,油酸可使SQS和CYP51两个灵芝三萜生物合成途径中的关键酶基因上调表达,从而使灵芝菌丝体胞内三萜产量大幅度提高。油酸对14α-LDM酶基因主要为抑制作用,从而抑制麦角甾醇这一代谢途径,间接提高灵芝三萜产量。     

L-缬氨酸生物合成的基础研究和菌种选育的进展

L-缬氨酸在医药、食品及饲料领域中有着广泛的用途.L-缬氨酸合成途径的调节有很多特点:比如该途径与L-异亮氨酸合成途径有多步反应共用同一种酶;该途径与另外两支链氨基酸合成途径是平行关系;该途径涉及的关键酶的活性和基因的表达都存在多价调节机制.本文概述了L-缬氨酸的生物合成途径关键酶的性质和基因表达的特点的研究进展;并结合上述机理阐述了国内外L-缬氨酸生产菌的传统的诱变育种方法和兴起的基因工程育种方法的发展现状.     

苯丙氨酸生物合成的整体代谢网络调控基因的敲除

csrA基因产物是大肠杆茵芳香族氨基酸生物合成途径中碳中心代谢有关的一种全局性调控蛋白质.采用Red敲除系统介导的同源重组的方法定位缺失大肠杆茵染色体csrA基因,经PCR、DNA测序等多种方法证实了基因剔除的可靠性.csrA基因剔除后,大肠杆菌芳香族氨基酸生物合成中受csrA表达产物负调控的关键酶的酶活力有所提高,而正调控的关键酶基因活性降低,突变菌株发酵生产苯丙氨酸的能力提高近13％.     

采后硅酸钠处理对厚皮甜瓜防卫基因mRNA的诱导表达

本文研究了采后100 mmol/L硅酸钠浸泡处理对甜瓜果实苯丙烷代谢途径关键酶苯丙氨酸解氨酶(PAL)、查尔酮合成酶(CHS)、及抗病相关酶过氧化物酶(POD)、β-1,3-葡聚糖酶基因(β-1,3-Glu)mRNA表达的影响。结果表明:硅酸钠处理可提高PAL、CHS、POD、β-1,3葡聚糖酶基因mRNA的表达。其中经硅酸钠处理后PAL、CHS、POD基因分别于6、6、2 d达到最大表达量,其表达量分别为对照组的2.69、2.48、3.77倍。由此表明,采后硅酸钠处理厚皮甜瓜果实可通过诱导苯丙烷代谢途径关键酶PAL、CHS,及抗病相关酶POD、β-1,3葡聚糖酶基因的转录增强采后抗病性。     

内酯类化合物代谢中的微生物酶及其应用

内酯类化合物在活生物体内经酶的作用产生代谢变化,可生成有利于生物体利用的物质.文中概述了近年来报道的几种有关内酯类化合物代谢中的微生物酶及其在生物技术中的应用情况和研究进展.     

火焰原子吸收光谱法测定白酒中锰含量的探讨

利用火焰原子吸收光谱法,对白酒直接进样,白酒挥去醇及酯类与水定容、白酒挥去醇及酯类与酸定容、白酒挥去醇及酯类经消解后与水定容和白酒挥去醇及酯类经消解后与酸定容5种制样方法获得的酒样和模拟酒样进行测定,结果表明:白酒中醇及酯类物质对锰含量测定影响较大,低浓度硝酸和消解对锰含量测定影响较小,白酒采用挥去醇及酯类直接与水定容测定其中的锰含量为一种简便、可行、快速的测定方法.     

Function and regulation of yeast genes involved in higher alcohol and ester metabolism during beverage fermentation

The biochemical formation of yeast-derived sensory-active metabolites like higher alcohols and esters determines the different characteristics of aroma and taste in fermented beverages. In yeast fermentation process, a large number of environmental factors affecting the production of volatile aroma compounds are abundant. Factors like substrate composition in fermentation media as well as process parameters influencing these flavor-active metabolites have already been described. These factors can act on the expression of yeast genes involved in aroma metabolism resulting in concentration differences in esters and higher alcohols important for flavor and taste. The understanding of the function of genes involved in biosynthetic pathways of aroma-active substances as well as their regulatory mechanisms is needed to control the production of ester and higher alcohol synthesis to create specific aroma profiles in fermented beverages. This review discusses the known regulation and function of several individual genes (ATF1, ATF2, EEB1, EHT1, BAT1, BAT2 and BAP2) described in fusel alcohol and ester synthesis mainly in S. cerevisiae and S. pastorianus var. carlsbergensis. Also, different factors like oxygen and temperature that allow ester and higher alcohol synthesis to be controlled during yeast fermentation are described.     

基于LOX-HPL途径的武夷肉桂加工中香气物质的形成与调控

武夷肉桂是以福建省级茶树良种肉桂(Camellia sinensis cv.Rougui)鲜叶为原料制成的乌龙茶,具有"岩骨花香"的品质特征,深受消费者青睐。除茶树鲜叶外,武夷肉桂的品质形成很大程度上取决于加工工艺,但生产加工过程中以绿叶挥发物(Green Leaf Volatiles,GLVs)和果酯类为代表的脂肪族类香气成分代谢机理的研究,尚未见报道。以武夷肉桂的鲜叶、晒青叶及做青过程叶为试材,采用顶空固相微萃取法(Headspace Solid-Phase Microextraction,HS-SPEM)结合气相色谱与高通量飞行时间质谱联用技术(Gas Chromatography-Time-of-Flight Mass Spectrometry,GC-TOF-MS)对加工过程样中的脂肪族类香气成分检测发现:脂肪族醛类在做青过程中显著降低,而大量的脂肪族醇类和酯类显著积累,其中叶醇酯类和己烯醇、酯类分别对静置失水和机械损伤存在响应;利用紫外/可见分光光度法检测加工过程样中LOX-HPL途径中关键酶活力,结果表明,脂肪氧合酶(Lipoxygenase,LOX)、乙醇脱氢酶(Alcohol dehydrogenase,ADH)及乙醇酰基转移酶(Alcohol acyltransferases,AAT)的活力分别在第三次摇青叶(3T)、第四次摇青后叶(4T)及第五次摇青后叶(5T)中达到最大值;实时荧光定量聚合酶链式反应(Quantitative Real-Time Polymerase Chain Reaction,qRT-PCR)结果表明,CsLOX2、CsLOX5基因主要在做青初始阶段(2T、3T)相对表达量较高,而随着工艺过程的推进,CsADH2-1、CsAAT基因在做青后期(4T、5T)开始上调表达;通过Pearson系数评估武夷肉桂加工过程中的脂肪族类香气、酶活力和基因表达量间的相关性,发现正己醇、戊酸叶醇酯、己酸己酯、丁酸甲基叶醇酯、柳酸叶醇酯等大量具有天然花果香的脂肪族醇类和酯类与CsADH2-1基因具有显著(P<0.05)或极显著(P<0.01)的正相关,CsAAT基因与CsLOX2、CsLOX5基因在加工过程中具有相似的表达模式(r=0.762,r=0.732)。LOX-HPL途径关键基因的共表达,驱动了脂肪族类香气在加工过程中的协同积累,是武夷肉桂馥郁香气形成的重要基础。     

Influence of carbon and nitrogen source on production of volatile fragrance and flavour metabolites by the yeast Kluyveromyces marxianus

The yeast Kluyveromyces marxianus produces a range of volatile molecules with applications as fragrances or flavours. The purpose of this study was to establish how nutritional conditions influence the production of these metabolites. Four strains were grown on synthetic media, using a variety of carbon and nitrogen sources and volatile metabolites analysed using gas chromatography–mass spectrometry (GC–MS). The nitrogen source had pronounced effects on metabolite production: levels of the fusel alcohols 2‐phenylethanol and isoamyl alcohol were highest when yeast extract was the nitrogen source, and ammonium had a strong repressing effect on production of 2‐phenylethyl acetate. In contrast, the nitrogen source did not affect production of isoamyl acetate or ethyl acetate, indicating that more than one alcohol acetyl transferase activity is present in K. marxianus. Production of all acetate esters was low when cells were growing on lactose (as opposed to glucose or fructose), with a lower intracellular pool of acetyl CoA being one explanation for this observation. Bioinformatic and phylogenetic analysis of the known yeast alcohol acetyl transferases ATF1 and ATF2 suggests that the ancestral protein Atf2p may not be involved in synthesis of volatile acetate esters in K. marxianus, and raises interesting questions as to what other genes encode this activity in non‐Saccharomyces yeasts. Identification of all the genes involved in ester synthesis will be important for development of the K. marxianus platform for flavour and fragrance production. Copyright © 2014 John Wiley & Sons, Ltd.     

预糖化米酒发酵工艺研究

针对传统米酒发酵工艺的创新升级以及大罐发酵的需求,对米酒发酵过程大米熟料与生料预糖化发酵等进行了深入探究。结果表明,原料经过蒸煮糖化后发酵,发酵醪还原糖含量升高至2倍左右,蒸馏液酒精度提升了10.62%,酒精度达到接近峰值的时间提前1～2 d,但是总酸、总酯含量都有所降低;熟料制浆糖化后酒精度进一步提高9.31%,总酯含量提高56.71%,但总酸略有降低;生料粉碎糖化酒精度进一步提升8.61%,但总酯含量相对减少21.47%。生料粉碎糖化发酵节省能源,发酵醪分散均匀、流动性好,在大罐发酵方面有着广阔的应用前景。     

Gene expression in wheat beer yeast strains and the synthesis of acetate esters

The synthesis of aroma compounds represents one of the most important parameters in beer production. Although it has been a historical topic of research, exactly how aroma components are formed has yet to be fully explained. Moreover, all of the research that has been published on yeast strains is focused on lagers and ales. Wheat beer yeast strains have not been the focus of aroma and flavour research. In this study, five different wheat beer yeasts were analysed to determine their capacity for producing acetate esters. In this study, the most commonly used wheat beer yeast strains for the production of German‐style wheat beer were analysed. This involved measuring the level of expression of the alcohol acetyl transferase genes ATF1, ATF2 and IAH1 over a period of 4 days (during primary fermentation) and plotting the data to observe the development of expression of the genes over time. Results confirmed their capacity to form acetate esters and showed a distinct correlation with increasing expression of the gene ATF1. However, the findings also indicated that gene expression in different yeast strains can vary considerably during fermentation. Copyright © 2016 The Institute of Brewing & Distilling     

两种方法提取丢糟中酯类成分的比较研究

采用CO2超临界萃取法和60％酒精热浸提法提取丢糟中的酯类物质，气相色谱分析结果表明：①两种方法均未能提取丢糟中的乙酸乙酯；②CO2超临界萃取法能够提取丢糟中的乳酸乙酯、己酸乙酯、丁酸乙酯和戊酸乙酯，而酒精热浸提法只能提取出乳酸乙酯和戊酸乙酯，且后者乳酸乙酯的含量达275．9285mg／100g，远远超过了前者的11．8955mg／100g。因此，提取丢糟中主要酯类成分，CO2超临界萃取法优于60％（v／v）酒精热浸提法。     

DISTRIBUTION OF ESTERS PRODUCED DURING SUGAR FERMENTATION BETWEEN THE YEAST CELL AND THE MEDIUM

The distribution of the esters formed during sugar fermentations between the yeast cells and the medium was investigated in fermentations by 5 strains of Saccharomyces cerevisiae and 3 strains of S. uvarum (carlsbergensis). The esters studied included the acetates of isoamyl alcohol and phenethyl alcohol and the ethyl esters of the C₆-C₁₂ fatty acids. All of both acetates appeared in the medium. The proportion of the fatty acid ethyl esters transferred to the medium decreased with increasing chain length: all in the medium for ethyl caproate, 54–68% for ethyl caprylate, 8–17% for ethyl caprate, and all remaining in the yeast cell for ethyl laurate. A higher proportion of the esters formed appeared to remain in the cells of the S. uvarum strains than in cells of S. cerevisiae.     

草莓、柿子复合果酒发酵过程中香味成分比较分析

采用二氯甲烷萃取法和气相色谱-质谱联用技术(GC/MS)测定草莓、柿子复合果酒发酵过程中第0、4、7、17、27、37 d的香味成分,使用峰面积归一法计算各成分的相对含量。结果表明:复合果酒在发酵过程中共检测出香味成分63种,其中12种酯类,11种醇类,10种酸类,5种酮类,13种烷烃类,12种其它成分,其中正戊醇、异戊醇、苯乙醇、亚油酸和对羟基苯乙醇等香味成分的含量较高,含量最高时分别为57.68%、45.67%、38.06%、17.97%和4.81%。随着发酵的进行香味成分的种类数量随之增多,酯类、酮类物质相对含量呈上升趋势,醇类物质呈先下降后上升趋势,酸类物质呈先上升后下降趋势,其中醇类物质相对含量最高,酯类物质次之,发酵至37 d各种香味成分已基本形成,已具有纯正、浓郁、优雅的果酒香气。