鸡蛋清中溶菌酶的提取与抑菌作用

研究从鸡蛋清中提取与精制溶菌酶的方法,观察溶菌酶对大肠杆菌与金黄色葡萄球菌的抑制作用.从鸡蛋清提取溶菌酶的方法:取出鸡蛋清,过滤,用蒸馏水溶解,加氯化钠,调节溶液pH至5.0,加入溶菌酶晶种,4℃下静置结晶,过滤,可以得到溶菌酶粗品.将粗品溶菌酶溶解,按上述步骤溶解后结晶,重复3次,可以精制溶菌酶.用滤纸片法观察溶菌酶对大肠杆菌与金黄色葡萄球菌的抑菌作用.结果表明,鸡蛋清溶菌酶提取的较优条件是:酶液pH 10.0、氯化钠加入量5%、蛋清液用2倍体积蒸馏水稀释.70℃下保温5min,鸡蛋清溶菌酶的精制率最高.鸡蛋清溶菌酶对大肠杆菌与金黄色葡萄球菌有明显抑制作用,80℃加热30min对鸡蛋清溶菌酶的抑菌作用影响很小.     

Influence of some oenological practices on the aromatic and sensorial characteristics of white Verdejo wines

Changes in the aromatic composition as well as sensory characteristics in Verdejo white wines were analysed based on two factors: the winemaking methodology and the storing time of wine in bottles. The volatile components were determined by GLC‐MS, and the sensory profile was designed and assessed according to the ISO 11035 standard. The results showed that when wines were made in oak barrels, either completely or partially, which means the wines were in contact with the lees, the levels of 1‐octanol, ethyl heptanoate and ethyl decanoate were significantly affected (P < 0.05); the softness sensation was also influenced (P < 0.05). However, the amount of time the wines were stored in bottles significantly affected (P < 0.05) the levels of 1‐hexanol, ethyl heptanoate, ethyl octanoate, ethyl decanoate, hexyl acetate, isoamyl acetate and isoamyl lactate and also an odour note (tropical fruit). The compounds with higher OAV values belong to the groups of esters and fatty acids. For these reasons, the composition and the quality of the aroma of Verdejo white wines appear to be significantly affected both by use of oak barrels in winemaking and the time the wines are stored in bottles.     

本土酿酒酵母的筛选及其低醇葡萄酒特性研究

以天津宝坻产区的三种葡萄为原料,分别从自然发酵的葡萄汁中筛选出340株酵母菌,经生理生化,产酸、酯、尿酶、H2S,发酵能力测定及26SrRNA测序,确定4株为酿酒酵母,用于贵人香低醇白葡萄酒的酿造。以商用酵母为对照,定量分析了不同酵母所酿酒中的香气成分、氨基甲酸乙酯含量,并进行了理化指标和感官测定,发现酵母C508和G611酿造的低醇酒具有更独特的香气特点,氨基甲酸乙酯的含量较低,微生物稳定性较高,更适合低醇酒的酿造。     

The effect of in-amphorae aging on oenological parameters,phenolic profile and volatile composition of Minutolo white wine

A wine was obtained from cryomacerated Minutolo grapes under reductive conditions and aged for 12 months in glass container and in 3 types of amphorae. After aging, wines in glass containers showed the highest alcohol content, volatile acidity, dissolved oxygen, concentrations of aromatics, alcohols, and esters and by the lowest contents of enols and terpenes. They also showed the highest decrease of flavonoids, hydroxycinnamoyl tartaric acids, and procyanidins. Wines in raw amphorae showed the dramatic decrease of flavonoids and flavans reactive with vanillin. The highest antioxidant activity was exhibited by wines in engobe amphorae, while the lowest values were showed by the wines in glass containers and glazed amphorae. Caftaric acid and procyanidin B3 decreased in wine aged under glass while epicatechin mainly reduced in raw amphorae.According to the Principal Component Analysis, the wines resulted homogeneously grouped as a function of the type of container in which were aged.     

Replacement of sulfur dioxide by lysozyme and oenological tannins during fermentation: influence on volatile composition of white wines

BACKGROUND: In recent years the use of sulfur dioxide, a commonly used additive in winemaking, has been questioned because of its toxic effects on human health. Studies have been conducted to find alternatives that can effectively substitute for this additive in all its various technological functions. In previous work, lysozyme and oenological tannins were found as possible substitutes in controlling bacterial undesirable fermentations and phenolic oxidation. However, data on the volatile composition of wines obtained by that protocol are lacking. In this work, the effects on volatile composition of white wines by the substitution of SO₂ during fermentation with lysozyme and tannin were studied. At the same time, the technological performance of two strains of yeast that produce low amounts of SO₂ were evaluated. RESULTS: The results showed that both SO₂ and lysozyme prevented the development of undesirable bacterial fermentations. The study of volatile compounds shows differences in the alcohol, acid and ester contents among the final products: wines fermented with strain 1042 and lysozyme had higher total alcohol concentration, while the addition of SO₂ promoted higher production of 3‐methyl‐1‐butanol, 3‐methylthio‐1‐propanol, phenylethyl alcohol and 4‐hydroxy‐benzenethanol. Esters, as a total, were influenced by the different strain and tannins addition, while amounts of medium‐chain fatty acid ethyl esters and their corresponding fatty acids were found in higher amounts in wines coming from fermentations with lysozyme. The sensory analysis revealed a preference for wines to which lysozyme and tannins had been added. CONCLUSION: The data suggest that the addition of lysozyme and oenological tannins during alcoholic fermentation could represent a promising alternative to the use of SO₂ and for the production of wines with reduced content of SO₂. The composition of the volatiles in the final wines was affected by the different vinification protocols (mainly with regards to alcohols and ethyl esters). Copyright © 2009 Society of Chemical Industry     

Biogenic amines in wines: Influence of oenological factors

Biogenic amines formation results from the decarboxylation of the corresponding amino acids by action of microorganisms. The presence of these compounds is considered by some authors a fundamental parameter for detriment of alcoholic beverages. The aim of this work was to assay the effect of some oenological factors (viticulture region, grape variety, anti-fungi treatment of grapes, fermentation activators, malolactic starters and storage on lees) from the point of view of their influence on the biogenic amines content of wines. According to our results, it was possible to show that the viticulture region affects the amounts of amines, since wines of some regions present higher contents of amines than wines from other regions. Grape varieties appear to influence the wine amines content. Commercial malolactic starters, after careful selection, should be added to the vinification process in order to decrease the formation of biogenic amines, since in our assays the wines that were inoculated with starters present lower amounts of biogenic amines. The wine storage on lees contributes for a biogenic amines increase.     

Fermentation of sulphite-free white musts with added lysozyme and oenological tannins: Nitrogen consumption and biogenic amines composition of final wines

The influence on amino acid consumption and biogenic amines composition of white wines obtained by replacing SO₂ during fermentation with lysozyme and tannins was studied. At the same time, the fermentative performance of two low SO₂ producing selected yeasts strains was evaluated. For this purpose, a series of laboratory-scale fermentations of fresh white must and a HPLC-DAD method for the analysis of amino acids, biogenic amines and ammonium ion were undertaken. The presence of SO₂ or lysozyme affected the consumption of nitrogen as a function of the yeast strain, while oenological tannin had no substantial influence. Strain 1042 increased the consumption of total YAN in the presence of SO₂, as a consequence of the enhanced utilization of ammonium ion and a number of amino acids. By contrast, strain 333 tended not to change the total YAN uptake, whatever the juice treatment, and reduced the consumption of aspartic and glutamic acids, GABA and other compounds in the case of samples added with SO₂. When compared with lysozyme addition, for both strains, SO₂ increased the consumption of alanine and glutamine, the latter being a major contributor to the assimilable nitrogen of the must. No influence of must treatments was found on the content of biogenic amines in the final wines.     

Production of indole by wine-associated microorganisms under oenological conditions

A high concentration of indole has been linked to ‘plastic-like’ off-flavour in wines, predominantly in wines produced under sluggish fermentation conditions. The purpose of this study was to determine the ability of yeast and bacteria to form indole and whether tryptophan was required for indole accumulation during winemaking. Wine-associated yeast and bacteria species (Saccharomyces cerevisiae, Saccharomyces bayanus, Candida stellata, Hanseniaspora uvarum, Kluyveromyces thermoloterans, Oenococcus oeni, Lactobacillus lindneri, Pediococcus cerevisiae and Pediococcus parvulus) were screened for their potential to generate indole during alcoholic or malolactic fermentation. Tryptophan was required for the accumulation of indole in chemically defined medium, and all yeast and bacteria fermentations were able to accumulate indole. C. stellata showed the greatest potential for indole formation (1033 μg/L) and among the bacteria, the highest concentration was generated by L. lindneri (370 μg/L). Whether primary fermentation is the principle cause of indole formation remains to be determined. We hypothesise that during an efficient fermentation, indole is removed through catabolic metabolism, but, when a sluggish fermentation arises, non-Saccharomyces species might produce excess indole that is still present by end of fermentation.     

Effect of oenological practices on microbial populations using culture-independent techniques

Sulphur dioxide (SO(2)) addition and yeast inoculation are well-established practices in winemaking for restricting the growth of indigenous yeasts and bacterial populations. The effect of these oenological practices on wine microbial populations has been evaluated using culture-independent methods. These are quantitative PCR (qPCR) for the enumeration of yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB), and PCR-DGGE to determine the yeast and bacteria species diversity. The PCR-DGGE method detected a low yeast and bacteria species diversity. On the contrary, the specificity of the primers designed for the qPCR allowed that minor microbial groups such as Hanseniaspora were accurately quantified regardless of a large presence of other microbial groups such as Saccharomyces. From an oenological point of view, inoculation increased the proportion of Saccharomyces vs. non-Saccharomyces in a shorter time. Hanseniaspora increased during the first phase and decreased during the latter phases of the process, especially in the sulphited fermentations. Both yeast inoculation and SO(2) kept the LAB populations at very low level, while the AAB populations were hardly affected by these two practices.     

Two-step chromatographic procedure for the preparation of hen egg white ovotransferrin

 A two-step chromatographic procedure for the preparation of hen egg white ovotransferrin by using an anion exchanger is described. The method consists of a preliminary step of frontal chromatography, also called displacement chromatography, on a Q Sepharose Fast Flow column. This simple and powerful chromatographic step enables the pre-purification of ovotransferrin that is further purified by a second conventional preparative chromatography performed on the same Q Sepharose FF column. The ovotransferrin obtained is found to be homogeneous in electrophoresis and analytical chromatography. Its purity rate is calculated at 97.5% and its amino acid composition is close to that obtained from the nucleotide sequence of ovotransferrin. Moreover, ovotransferrin is iron-free, thus suitable for studies in relation to its metal-binding activity. Received: 31 March 2000 / Revised version: 29 May 2000     

蛋清肽的功能特性及制备技术研究进展

蛋清蛋白经蛋白酶适度水解,生成肽的混合物,通过控制其水解度,可提高蛋白的吸收率,同时,生成的多肽混合物更有利于人体吸收,此外肽还具有许多特殊生理功能。综述了蛋清肽的降血压、清除自由基、提高免疫力等功能特性,并介绍了制备蛋清肽的酶的选择及水解度的测定等关键工艺,以期为相关研究提供参考。     

Multilocus sequence typing of oenological Saccharomyces cerevisiae strains

This study describes the application of a multilocus sequence typing (MLST) analysis for molecular discrimination at the strain level of Spanish wine yeast strains. The discrimination power of MLST is compared to mitochondrial RFLP analysis. Fragments of the ADP1, ACC1, RPN2, GLN4, and ALA1 genes were amplified by PCR from chromosomal DNA of 18 wine Saccharomyces cerevisiae strains. Ten polymorphic sites were found in the five loci analyzed showing 13 different genotypes, with 11 of them represented by only one strain. RFLP analysis of the same 18 wine yeast strains showed seventeen different mitochondrial patterns. Phylogenetic relationships among the strains analyzed, inferred by MLST data, showed wine isolates of S. cerevisiae as a rather homogeneous group. The discrimination potential of mitochondrial RFLP analysis was superior to the MLST scheme used in this work. However, MLST analysis allowed an easy construction of reliable phylogenetic trees. MLST analysis offers the possibility of typing wine S. cerevisiae strains simultaneously to the study of the genetic relationship among them.     

Study of the oenological characteristicsand enzymatic activities of wine yeasts

Seventy-four wine strains of theSaccharomycesgenus were tested to study their oenological characteristics and enzymatic activities. These strains were isolated from spontaneous microbiota taken from cellars in the Valdepeñas region (Spain) and identified by contour homogeneous electric field (CHEF) technique. When Cluster Analysis was applied to oenological properties, only SH₂production, tolerance to 12⁰of ethanol and synthesis of killer toxin contributed to the distribution of the strains in the different statistical groups. All the strains possessed esterase activity and some also showed protease and pectinase activities. None of them hadβ-D-glucosidase activity.     

Pre-fermentative replacement of sulphur dioxide by lysozyme and oenological tannins: Effect on the formation and evolution of volatile compounds during the bottle storage of white wines

In this work, the effects on the volatile profile of the pre-fermentative substitution of SO₂ with lysozyme and oenological tannins were studied in white wines. At the same time, in order to understand the changes of volatile compounds in SO₂-free wines, the evolution of volatiles was evaluated over 1 year of storage in bottles. For this purpose, a number of laboratory scale fermentations of SauvignonBlanc musts were carried out and the effects of three variables (SO₂, lysozyme and oenological tannins) were investigated by means of GC–MS analysis. Results showed that the replacement of SO₂ with lysozyme and oenological tannins influenced the volatile composition of wines at the end of the alcoholic fermentation. Wines fermented with SO₂ showed higher total alcohol amounts, while the presence of oenological tannins augmented the level of esters. The presence of SO₂ influenced also the alcohols and esters profiles of wines during bottle storage. Moreover, the presence of oenological tannins displayed a positive role in maintaining the amounts of esters over certain levels in wine stored for 1 year, likely due to their oxygen scavenging ability. By contrast, acids were less affected by the investigated adjuvants both at the end of the alcoholic fermentation and during the storage time.     

Production and characterization of anAspergillus terteus α-l-rhamnosidase of oenological interest

Conditions for maximal batch culture production of extracellularα-L-rhamnosidase byAspergillus terreus have been investigated. Production of the enzyme appeared to be inducible by rhamnose and rutin, reaching a maximal level after an incubation period of 162 h when the fungus was grown at 37°C on either of these compounds as the carbon source and on ammonium phosphate as the nitrogen source. Nonionic surfactants did not enhanceα-L-rhamnosidase secretion. Under optimal conditions,A. terreus produced only oneα-L-rhamnosidase of approximate molecular weight 90 kDa (SDS-PAGE) and isoelectric point of 4.6. Onp-nitrophenyl-α-L-rhamnopyranoside as substrate, the enzyme showed pH and temperature optima of 6–8 and 45–50°C, respectively. Neither divalent cations nor ethylenediaminetetraacetate (EDTA) inhibited or stimulated enzyme activity. The enzyme was active at the concentrations of glucose found in must or of ethanol in wine.     

Hygienic study of a proteolytic preparation obtained from Actinomyces]

An enzymatic preparation rimoprotelin, obtained from Actinomyces rumosus was studied. The results of a microbiological control analysis showed the preparation to contain admixtures of oxytetracycline in an amoun of 950 Un/g. In a 9-months long test with albino rats the enzymatic preparation produced substantial interferences in the composition to the intestinal microflora and increased the incidence of bacterial cultures resistent to oxytetracycline. Morphological changes in the organs and tissues of the test animals, which were, apparently, conditioned by the presence of the antibiotic admixture in the preparation were revealed. This point to the need of a thorough purification of this enzymatic preparation from the oxytetracycline admixture and makes its use as a food additive in its original form imposssible.     

Resurgence of minority and autochthonous grapevine varieties in South America: a review of their oenological potential

In contrast with the general trend of producing wine from the most famous grapevine varieties, associated with the French paradigm, such as Cabernet-Sauvignon, Merlot, Pinot Noir, Syrah, Sauvignon Blanc, and Chardonnay, there is a tendency to revalorize and preserve minority or autochthonous grapevine varieties worldwide. The South American wine region, where most of the varieties derived from varieties brought after European colonization, is not exempt from this. This has allowed new wines to be provided with distinctive identities that are markedly different from the current homogeneous wine production. Moreover, varietal homogenization increases vineyard genetic vulnerability in relation to the emergence of grapevine diseases, to which the commonly cultivated varieties are not resistant. This review summarizes the oenological potential of minority or autochthonous grapevine varieties cultivated within the South American wine region, focusing on Argentina, Chile, and Bolivia. © 2019 Society of Chemical Industry     

Genetic screening of lactic acid bacteria of oenological origin for bacteriocin-encoding genes

A total of 330 lactic acid bacteria isolated from South African red wines during alcoholic and malolactic fermentations and 9 commercial malolactic bacteria starter cultures were screened for antimicrobial activity. Of the entire screened isolates, 26 strains, belonging to the species Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus hilgardii and Oenococcus oeni, showed activity towards various wine-related and non-wine-related indicator strains. A PCR-based screening revealed the presence of the plantaricin encoding genes plnA, plnEF, plnJ and plnK in five selected Lb. plantarum strains. Furthermore, a co-culture experiment with Lb. plantarum and Enterococcus faecalis was performed. A complete inhibition of cell growth of Ent. faecalis was observed within 72 h. Four putative bacteriocin-encoding genes in the genome of O. oeni were identified and sequenced.     

中/欧葡萄酒工艺标准比较

为了进一步完善中国葡萄酒标准,促进中国葡萄酒的国际贸易,有必要对中国、欧盟现行的葡萄酒工艺标准进行比较研究。该文将GB 15037—2006《葡萄酒》、《中国葡萄酿酒技术规范》及GB 2760—2014《食品添加剂使用标准》等葡萄酒相关规定与欧盟葡萄酒相关标准进行对比分析。结果表明,中欧现行葡萄酒工艺标准的差异主要存在于以下方面:葡萄酒定义和分类、葡萄品种、葡萄酒工艺(特别是添加剂和加工助剂)以及有关物质的限量等。并为中国葡萄酒标准体系的修订提出建议和注意原则。