不同波长Ar^＋激光对啤酒酵母菌的诱变效应

本文应用514.5nm,496.5nm、488.0nm和476.5nm等四种不同波长的氩离子辐照啤酒酵母菌,激光辐照的功率密度300mw/cm~2、辐照时间12min。辐照后进行细胞培养和啤酒发酵试验,并测定细胞有关生理性能的变化。实验表明,上述四种波长的Ar~+激光对啤酒酵母细胞的生长繁殖和代谢主产物乙醇的生物合成有明显的促进作用,并能提高双乙酰还原酶的活力,降低发酵付产物双乙酰的含量。这些变化,有利于缩短啤酒发酵的周期,提高啤酒产量相改善啤酒的风味。     

Bestimmung von Deoxynivalenol in Brot und Bier

Analytical procedures for the determination of deoxynivalenol (DON) in bread and beer, using enzyme immunoassay (EIA) and HPLC methods, were developed. For determination of DON by EIA, aqueous raw extracts of bread or degassed beer were extracted by liquid-liquid partitioning with ethyl acetate, the organic solvent evaporated, and the residue redissolved in phosphate buffered saline (PBS) for analysis. For determination by HPLC (UV detection at 218 nm), DON in bread extracts or beer was purified on immunoaffinity chromatographic columns. In bread, detection limits for DON of 15 µg/kg (EIA) and 7 µg/kg (HPLC) were achieved, with mean recoveries of 81%. In beer, the detection limit for DON was 2 µg/l both in EIA and HPLC, with recoveries of 91–93%. Both methods showed good agreement of the results for naturally contaminated sample materials, with r²=0.993 for bread and r²=0.823 for beer, respectively.     

Intrinsic tryptophan fluorescence of human serum proteins and related conformational changes

The unfolding of human serum proteins (HSP) was studied by measuring the intrinsic fluorescence intensity at a wavelength of excitation corresponding to tryptophan's or typosine's fluorescence and surface hydrophobicity. The maxima emission wavelengths (_max) of human serum albumin (HSA) and human serum globulin (HSG) before beer consumption (BC) were 336.0 and 337.0 nm and after BC shifted to 335.0 and 334.0 nm, respectively. The surface hydrophobicity slightly increased after BC. In a solution of 8 M urea the _max of BSA shifted to 346.4 and that of BSG to 342.5 nm. In contrast, in the same solution but after BC the _max positions of HSA and HSG shifted to 355.9 and 357.7 nm, respectively. A decrease in fluorescence intensity, a shift in the maximum of emission, and an increase in surface hydrophobicity which reflected unfolding of proteins were observed. Here we provide evidence that the loosening of the HSP structure takes place primarily in various concentrations of urea before and after beer consumption. Differences in the fluorescence behavior of the proteins are attributed to disruption of the structure of proteins by denaturants as well as by the change in their compactability as a result of ethanol consumption.     

Genome sequence of the phage clP1, which infects the beer spoilage bacterium Pediococcus damnosus

Pediococcus damnosus (P. damnosus) bacteriophage (phage) clP1 is a novel virulent phage isolated from a municipal sewage sample collected in Southern Ireland. This phage infects the beer spoilage strain P. damnosus P82 which was isolated from German breweries. Sequencing of the phage has revealed a linear double stranded DNA genome of 38,013 base pairs (bp) with an overall GC content of 47.6%. Fifty seven open reading frames (ORFs) were identified of which 30 showed homology to previously sequenced proteins, and as a consequence 20 of these were assigned predicted functions. The majority of genes displayed homology with genes from the Lactobacillus plantarum phage phiJL-1. All genes were located on the same coding strand and in the same orientation. Morphological characterisation placed phage clP1 as a member of the Siphoviridae family with an isometric head (59 nm diameter) and non-contractile tail (length 175 nm; diameter 10nm. Interestingly, the phage clP1 genome was found to share very limited identity with other phage genome sequences in the database, and was hence considered unique. This was highlighted by the genome organisation which differed slightly to the consensus pattern of genomic organisation usually found in Siphoviridae phages. With the genetic machinery present for a lytic lifecycle and the absence of potential endotoxin factors, this phage may have applications in the biocontrol of beer spoilage bacteria. To our knowledge, this study represents the first reported P. damnosus phage genome sequence.     

啤酒生产酵母全循环新工艺的研究

以改善浅色啤酒质量,降低生产成本,提高经济效益的目的,提出一种新颖的浅色啤酒的酿造方法。采用煮－浸法糖化工工艺及５０％麦芽和５０％大米作为啤酒酿造的原料及辅料。在糖化过程中添加啤酒酵母提取物作为补充氮源,不仅使所酿造的成品啤酒色泽浅,口味淡爽,纯正,泡沫洁白细腻,持久挂杯,而且具有较显著的经济效益和社会效益。     

Characterization of new 18-kDa IgE-binding proteins in beer

The IgE-binding proteins in beer were examined by immunoblotting analysis with sera of patients sensitive to beer. Several proteins were immunoblotted with the sera, and among these, 18-kDa proteins were identified as new IgE-binding proteins in beer. Perhaps they originated from barley as a raw material.     

The influence of alcohol-containing and alcohol-free beverages on lipid levels and lipid peroxides in serum of rats

It is an established fact that moderate consumption of alcoholic beverages leads to some positive biochemical changes in blood that are widely regarded as indicators of improved prevention of atherosclerosis. However, at present, there are different opinions regarding the biologically active compounds of alcoholic beverages that bring about these changes. This experiment was conducted on 60 male Wistar rats, which were divided into five groups, each of which contained 12 rats: four experimental groups (EG1, EG2, EG3, EG4) and one control group (CG). During 4 weeks, all groups of rats were fed basal diet (BD) supplemented with dry red wine (EG1), beer (EG2), lyophilized dry red wine (EG3), or lyophilized beer (EG4). The rats of the CG were fed BD only. The rats of EG1 and EG2 were fed BD supplemented daily with 2.0 mL of wine and 6.0 mL of beer, respectively. The rats of EG3 and EG4 were fed BD supplemented daily with lyophilized wine and lyophilized beer at a concentration corresponding to an intake of 2.0 mL of original wine and 6.0 mL of original beer, respectively. Before and after completion of the trial, a wide range of laboratory tests including lipids and lipid peroxides were performed. The results of this investigation reveal that both original and lyophilized wine and beer exercise statistically significant beneficial lipidemic and antioxidant effects by reducing total cholesterol (TC), low density lipoprotein cholesterol, triglycerides, and lipid peroxides (P < 0.05 for all) and by elevating the high density lipoprotein cholesterol:TC ratio. There were no statistically significant differences in the results between groups fed BD supplemented with original wine and beer versus groups fed BD supplemented with lyophilized wine and beer. Therefore, it can be concluded that the biologically active compound of these beverages is their dry matter containing inter alia polyphenols in relatively high concentrations.     

Removal of diacetyl from beer by adsorbants and diacetyl reductase

Removal of diacetyl from beer with adsorbants like cellulose, silica gel, activated charcoal, calcium phosphate gel, anion- and cation-exchange resins, and silicylic acid black soil bed (SABSB) was attempted in comparison with the enzyme diacetyl reductase (EC 1.1.1.5). Diacetyl could be removed from beer by the adsorbants but they had undesirable effect on the beer quality such as color, pH, and alcohol levels. These adverse effects were not observed with the use of diacetyl reductase. The results favor the enzymatic removal of diacetyl from beer as a superior approach.     

Determination of D-amino acids labeled with fluorescent chiral reagents, R(-)- and S(+)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N, N-dimethylaminosulfonyl)-2,1,3-benzoxadiazoles, in biological and food samples by liquid chromatography

D-Amino acids in food and biological samples labeled with R(-)- and S(+)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N, N-dimethylaminosulfonyl)-2,1,3-benzoxadiazoles (DBD-PyNCS) were separated by reversed-phase chromatography and detected fluorometrically at 550 nm (excitation at 460 nm). DL-Amino acids were efficiently labeled at 55 degrees C for 20 min in basic medium. The resulting thiocarbamoyl-amino acids were resolved by an isocratic elution using water:30% methanol in acetonitrile (72:28) containing 0.1% trifluoracetic acid as mobile phase for hydrophilic amino acids and gradient elutions using sodium acetate buffer (pH 5. 2)/acetonitrile as gradient solvent mixture for hydrophobic amino acids, respectively. The detection limits (S/N = 3) of DL-amino acids tested were in the range of 0.16-0.75 pmol. The proposed method was applied to determine the D-amino acid(s) in milk, cream, fermented dairy products (yogurt and yakult), tomato products (juice, puree, and catchup), fermented beverages (beer and red wine), and human urine. The existence of D-amino acid(s) was demonstrated in all the samples tested. Furthermore, the identification of the D-amino acid(s) was performed using both isomers of DBD-PyNCS and by on-line HPLC-electrospray ionization-MS.     

Application of chitooligosaccharides as antioxidants in beer to improve the flavour stability by protecting against beer staling during storage

Objective

To improve beer flavour stability by adding chitooligosaccharides that prevent formation of staling compounds and also scavenge radicals in stale beer.

Results

Chitooligosaccharides, at 0.001–0.01%, inhibited the formation of staling compounds in forced aged beer. The formation of 5-hydroxymethylfurfural, trans-2-nonenal and phenylacetaldehyde were decreased by 105, 360 and 27%, respectively, when compared with those in stale beer without chitooligosaccharide addition. The capability of chitooligosaccharides to prevent staling compound formation depended on their molecular size (2 or 3 kDa). The DPPH/hydroxyl radical scavenging activity in fresh beer significantly lower than that in forced aged beer in the presence of chitooligosaccharides. When compared with stale beer without added chitooligosaccharides, the radical scavenging activity could be increased by adding chitooligosaccharides to forced aged beer.

Conclusions

Chitooligosaccharides play an active part in the prevention of beer flavour deterioration by inhibiting the formation of staling compounds and increasing radical scavenging activity.

     

A study of the beer chill-proofing behaviour of a water-insoluble papain conjugate of hydrous titanium(IV) oxide-use of hydrous titanium(IV) oxide as a novel chill-proofing agent

An active papain (EC 3.4.22.2) conjugate of hydrous titanium(IV) oxide has been found to exhibit substantial ability to chill-proof beer. However, this ability has been shown to be nearly identical to the chill-proofing abilities found to be exhibited by an inactive S-carboxymethyl derivative of the papain conjugate, which was prepared by treating the papain conjugate with bromoacetic acid, and by free hydrous titanium(IV) oxide. Further, the chill-proofing achieved by each material was observed to correlate with reductions in the absorbance of beer in both the ultraviolet and visible regions of the spectrum. On this basis, it is concluded that chill-proofing by these materials occurs solely by the adsorption of beer constituents and that this adsorption is probably non-specific. The rider to this conclusion is that the coupling of papain to hydrous titanium(IV) oxide, in order to give an active immobilized enzyme that acts as a reusable chill-proofing agent, is without efficacy. The broader significance of these observations in the development of enzymic chill-proofing agents is discussed. The potential of free hydrous titanium(IV) oxide as an adsorbent chill-proofing agent has been examined. It has been shown that the treatment of beer for 12 h at 4°C with fresh hydrous titanium(IV) oxide (200 g/hl) produces nearly complete chill-proofing. Chill-proofing is enhanced both by prolongation of treatment and by increased number of treatments. Further, beer that had been chill-proofed by hydrous titanium(IV) oxide was found to contain titanium(IV) at a concentration below the detection limit (2 p.p.m.) of the adopted colorimetric method of analysis. These results auger well for the safe commercial use of hydrous titanium(IV) oxide as a novel and effective chill-proofing agent, particularly as hydrous titanium(IV) oxide may be prepared conveniently on site from a readily available and inexpensive material, titanium(IV) chloride.     

Absence of fks1p in lager brewing yeast results in aberrant cell wall composition and improved beer flavor stability

The flavor stability during storage is very important to the freshness and shelf life of beer. However, beer fermented with a yeast strain which is prone to autolyze will significantly affect the flavor of product. In this study, the gene encoding β-1,3-glucan synthetase catalytic subunit (fks1) of the lager yeast was destroyed via self-clone strategy. β-1,3-glucan is the principle cell wall component, so fks1 disruption caused a decrease in β-1,3-glucan level and increase in chitin level in cell wall, resulting in the increased cell wall thickness. Comparing with wild-type strain, the mutant strain had 39.9 and 63.41 % less leakage of octanoic acid and decanoic acid which would significantly affect the flavor of beer during storage. Moreover, the results of European Brewery Convention tube fermentation test showed that the genetic manipulation to the industrial brewing yeast helped with the anti-staling ability, rather than affecting the fermentation ability. The thiobarbituric acid value reduced by 65.59 %, and the resistant staling value increased by 26.56 %. Moreover, the anti-staling index of the beer fermented with mutant strain increased by 2.64-fold than that from wild-type strain respectively. China has the most production and consumption of beer around the world, so the quality of beer has a significant impact on Chinese beer industry. The result of this study could help with the improvement of the quality of beer in China as well as around the world.     

Development of engineered yeast for biosorption of beer haze-active polyphenols

Compared to most other alcoholic beverages, the shelf life of beer is much more limited due to its instability in the bottle. That instability is most likely to appear as turbidity (haze), even sedimentation, during storage. The haze in beer is mostly caused by colloidal particles formed by interactions between proteins and polyphenols within the beer. Therefore, beers are usually stabilized by removing at least one of these components. We developed and constructed a Saccharomyces cerevisiae strain with a proline-rich QPF peptide attached to the cell wall, using the C-terminal anchoring domain of α-agglutinin. The QPF peptide served to bind polyphenols during fermentation and, thus, to decrease their concentration. Strains displaying QPF were able to bind about twice as much catechin and epicatechin as a control strain displaying only the anchoring domain. All these experiments were done with model solutions. Depending on the concentration of yeast, uptake of polyphenols was 1.7–2.5 times higher. Similarly, the uptake of proanthocyanidins was increased by about 20 %. Since the modification of yeasts with QPF did not affect their fermentation performance under laboratory conditions, the display of QPF appears to be an approach to increase the stability of beer.

     

Immobilized pepsin: Properties and use to prevent haze formation in beer and wine

Summary The characteristics of a new immobilized pepsin preparation are shown as a function of pH-value and temperature. Optimal working conditions for beer and wine treatment are derived from the stability and activity data. It was found that good protein stability of the beverage results when at least 50 g immobilized enzyme per hl beer per h (alternatively 20 g per hl wine per hour) are applied in a special type of packed bed reactor. Since the foam stability of beer is decreased by the enzyme treatment, better perspectives for industrial application are seen in wineries than in breweries.     

几株啤酒酵母的筛选及发酵性能比较*

啤酒酵母是啤酒酿造的灵魂,可以直接影响啤酒品质。在啤酒酿造过程中,由于啤酒酵母被多次传代和保藏,造成优良菌种发酵性能衰退等问题,导致发酵不彻底,影响最后啤酒的风味质量。为此以8株Lager型啤酒酵母为出发菌株,通过平板分离纯化获得80株分离菌株,再经过三角瓶发酵初筛和复筛、发酵罐中试发酵实验最终获得了8株发酵性能优良的啤酒酵母。其中,6株酵母可应用于酿造双乙酰含量低于0.1 mg/L的啤酒;3株酵母发酵度高于70%,适合酿造干啤酒;1株酵母发酵度低于50%,适合酿造低醇啤酒。在风味方面:1株酵母酿造的啤酒醇酯比为3.3,啤酒酯香味较突出;另1株酵母酿造的啤酒醇酯比为4.5,啤酒高级醇含量较高。8株经过选育的啤酒酵母发酵特征明显,便于精酿啤酒厂实际应用。     

Reduced urination rate while drinking beer with an unpleasant taste and off-flavor

A lowered subjective evaluation of the taste and flavor of beer due to staleness or to the addition of an unpleasant taste and flavor was found to be closely correlated with the urination rate. Beer in the same lot was compared immediately after shipment from the brewery and after leaving at room temperature for 1 month or 5 months. Each beer sample was given to volunteers at the rate of 3 ml/kg/15 min for 2 hours, and the urine volume was measured every 30 minutes. The urination rate was highest from the volunteers who drank fresh beer and lowest from those who drank 5-month-old beer. The subjective evaluation of both the taste and drinkability of 5-month-old beer was significantly lower than that of fresh beer. Beer samples with various unpleasant taste and flavor substances added lowered the urination rate. The results suggest that the perception of an unpleasant taste and off-flavor would lower the urination rate.     

ILV2基因缺失对啤酒酵母生长与双乙酰代谢的影响

【目的】旨在应用分子生物学方法降低啤酒发酵液中双乙酰含量,改善啤酒感官质量。【方法】以酿酒酵母S2(Saccharomyces cerevisiae)为出发菌株,通过同源重组敲除四倍体啤酒酵母α-乙酰乳酸合成酶部分基因(ILV2),构建缺失一个和两个ILV2等位基因的突变株QI2-1和QI2-2,并进行啤酒发酵实验。【结果】ILV2基因的缺失,会导致菌株初始生长速率的降低。其中QI2-2较为明显,12 h时,突变株与出发菌株的生长速率达到一致。啤酒发酵结果表明,与出发菌株相比,突变株QI2-1双乙酰峰值与双乙酰最终含量分别降低17.50%和17.83%,而QI2-2分别降低51.67%和45.65%。其他啤酒指标如酒精度、发酵度、残糖和风味物质等略有变化,但都在优质啤酒指标范围内,符合啤酒发酵的质量要求。【结论】通过同源重组敲除部分ILV2基因和选育低产双乙酰菌株是降低啤酒双乙酰含量、提高啤酒质量的有效方法,具有一定的实际应用价值。     

Structure of Heliangine,a Novel Ten-membered Sesquiterpene Lactone

Determination of the I.T.T. (Indicator Time Test) value of beer has so far been conducted by the visual measurement. However, this visual judgement of the critical point, at which 80% of added dyestuff is accurately decolorized, is very difficult. We attempted to estimate the I.T.T. value more correctly using a spectrophotometer. On the basis of the following facts and observations, spectrophotometric determination of the I.T.T. value has been proposed. The absorption maximum of 2,6-dichlorophenolindophenol was observed at 520 mμ, but the absorption of beer was hardly ever recognized at this wave length. Walpole’s buffer solution was used for stabilizing the dyestuff at a pH similar to that of beer. Ethanol contained in beer did not affect the absorption of dyestuff. It was also found that the optical density of the dyestuff was proportional to the concentration both in water and in beer.     

Detection of resistance of lactic acid bacteria to a mixture of the hop analogue compounds tetrahydroiso-alpha-acids by noninvasive measurement of intracellular pH

AIMS: To examine the resistance of beer isolates of lactic acid bacteria (LAB) towards a mixture of tetrahydroiso-alpha-acids (Tetra) by growth experiments as well as by measurement of intracellular pH. METHODS AND RESULTS: Beer LAB isolates were identified to species level by SDS-PAGE of whole-cell proteins. Beer isolates of Lactobacillus brevis showed better ability for growth in the presence of Tetra than nonbeer isolates of the L. brevis or other species of LAB including beer and nonbeer isolates. The antimicrobial effect of Tetra was also examined by noninvasive measurement of intracellular pH by fluorescence ratio imaging microscopy for selected beer isolates of L. brevis and Pediococcus inopinatus. Strains of L. brevis showing limited decrease of intracellular pH during exposure to Tetra also showed better ability for growth in the presence of these compounds as well as in commercial beer products. CONCLUSIONS: It was possible to apply a method for noninvasive measurement of intracellular pH to predict the resistance of beer spoilage LAB towards the Tetra hop analogue compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the usability of a new rapid method for detecting hop-resistant variants of known beer spoilage LAB species.     

Significant abilities of titanium(IV)-activated glass fibre paper and its papain conjugates to chill-proof beer

It has been demonstrated that titanium(IV)-activated glass fibre paper, unlike untreated glass fibre paper, acts as a potent agent for chill-proofing beer: three treatments of beer with titanium(IV)-activated glass fibre paper (0.2m²/l) for 12 h were found to produce complete chill-proofing. Chill-proofed beer was found not to be contaminated with titanium(IV) species at a concentration above a detection limit of 2 p.p.m., so augering well for the safety of titanium(IV)-activated glass fibre paper as a chill-proofing agent. The physical form of titanium(IV)-activated glass fibre paper provides a major advantage over particulate chill-proofing agents in that it may be easily and rapidly separated from beer. The chill-proofing ability of titanium(IV)-activated glass fibre paper was found to decline with repeated use of the material. Moreover, all the observed chill-proofing effects were paralleled by reductions in the absorbance of beer across the ultraviolet and visible regions of the spectrum. These results demonstrate that titanium(IV)-activated glass fibre paper imitates hydrous titanium(IV) oxide in its effects upon the properties of beer and accord with other evidence that the surface of titanium(IV)-activated glass fibre paper is chemically similar to that of hydrous titanium(IV) oxide. The mechanism of the chill-proofing action of titanium(IV)-activated glass fibre paper is concluded to be analogous to that of hydrous titanium(IV) oxide and probably to involve the non-specific adsorption or ligand bonding of beer proteins and polyphenols to titanium(IV). Two papain (papain, EC 3.4.22.2) conjugates of titanium(IV)-activated glass fibre paper, of different catalytic activities, have been examined for chill-proofing ability. The chill-proofing behaviour observed for each papain conjugate was found not to differ from that observed for the corresponding catalytically-inactive S-carboxy-methyl derivative, prepared by treatment of the papain conjugate with bromoacetic acid, nor to differ greatly from that of titanium(IV)-activated glass fibre paper. On this basis the chill-proofing abilities of the papain conjugates are attributed solely to their abilities to adsorb beer constituents and not to their catalytic activities.