酒酒球菌在青梅汁中的生长及苹果酸乳酸发酵特性的研究

以青梅汁为原料,利用酒酒球菌的苹果酸-乳酸发酵（MLF）对青梅汁进行生物降酸的研究。结果表明:接种量为2.0×108CFU/mL,青梅汁的pH≥3.4时,MLF能正常进行,并使样品的酸度降低61.35%以上;接种量为2.6×107CFU/mL时,青梅汁的pH为3.6才能触发MLF,样品的酸度降低了77.60%;接种量为2.3×108CFU/mL,在pH3.6和4.0的青梅汁中,添加1.0g/100g的葡萄糖的样品,与不添加葡萄糖的样品相比,其酸度分别降低了27.04%和34.63%;在柠檬酸-柠檬酸钠缓冲体系中,酒酒球菌使体系酸度降低了26.40%,证实酒酒球菌可利用柠檬酸作为碳源,进行MLF。      

Use of a polymerase chain reaction for specific detection of the malolactic fermentation bacterium Oenococcus oeni (formerly Leuconostoc oenos) in grape juice and wine samples

A PCR method has been developed that enables rapid and direct identification of the malolactic bacterium Oenococcus oeni from grape must or wine samples. Two primers, based on unique, highly conserved regions within the 16S rRNA gene of O. oeni , were used to amplify a 995 bp fragment which is specific for O. oeni . Other species of bacteria from Lactobacillus, Pediococcus and Acetobacter which may be found in grape must or wine were not detected using this technique. This diagnostic test is able to specifically detect in the order of 10³colony forming units per mL of O. oeni in a wine sample, and can be used for monitoring bacterial growth during malolactic fermentation.     

Clarification of apple juice by electroflotation

Apple Juice industry is in search of a simplified technology which enables a quick clarification and stabilisation of apple juice. This study aimed to evaluate the potential of electroflotation as an alternative for the clarification of apple juice. Clarification of apple juice by electroflotation was first done at various current densities (10, 20 and 40 mA/cm²) with and without addition of gelatin (200 mg/l). Afterwards, the electroflotation treatments were done at a current density of 20 mA/cm² with various concentrations of added gelatin (0, 50, 100 and 200 mg/l). It was shown that electroflotation treatments alone was efficient to reduce the tannin and protein contents of apple juice. However, the decrease in the protein content was in large part due to the use of pectinases prior to the electroflotation treatments. The use of gelatin in combination with the electroflotation aided in the clarification process. The highest gelatin concentration used in this study (200 mg/l) resulted in a better reduction of tannin and protein levels, while a current density of 20 mA/cm² was found to be optimal. Turbidity observed in the juices clarified with electroflotation treatments was in average lower than 10 NTU but higher than 2 NTU which is generally required to produce a stable clarified juice. Brix degree and pH of the apple juice was not affected by the electroflotation treatments while the color was improved.

Industrial relevance

The production of clarified and stable apple juice is a subject of interest for the beverages industries. The clarification step which remained long and discontinuous implied the addition of a large quantity of pectolytic enzyme and of clarifying agents (such as gelatin) to the freshly pressed juice to induce the precipitation of proteins and other suspended matter in 15–20 h. Fining treatments were followed by a separation step usually consisting of decantation and classical filtration on filter-press, or flotation by dispersed gas. The development of membrane separation processes to replace the traditional approach has enabled the automation of the whole production resulting in lower labor requirement and a considerably shorter process time than the traditional process.However, the performance of membrane separation processes is influenced by the declining permeate flux with time, which is due to membrane fouling. In some instances, permeate flux decline makes membrane separation processes unattractive for the clarification of apple juice. To our knowledge, we are the first research group to use electroflotation (EF) for clarification of apple juice. It was shown that EF treatments alone were efficient to reduce the tannin and protein contents of apple juice. In addition, the use of gelatin in combination with the EF aided in the clarification process. Turbidity observed in the juices clarified with EF treatments for 30 min was in average lower than 10 NTU. Brix degree and pH of the apple juice were not affected by the EF treatments while the color was improved.When compared to the values reported in the literature for flotation by dispersed gas, it seems that EF shows better efficiency than flotation in decreasing the juice turbidity (99% decrease for EF as compared to 90% decrease for flotation). In addition, for experiments carried out by conventional flotation larger amount of fining agent are used (70–150 mg of gelatin/l, 400–800 mg/l of silica sol and 200–500 mg/l of bentonite). For these reasons, the new process we propose is advantageous when compared to the traditional flotation approach and it should have a measurable impact on the advancements in the production of clarified apple juice. If used as a pre-treatment to ultrafiltration clarification, it is expected that it would reduce membrane fouling resulting in higher productivity.     

Oenococcus oeni,a species born and moulded in wine: a critical review of the stress impacts of wine and the physiological responses

Oenococcus oeni is a lactic acid bacterium highly adapted to the stressful environment of wine, and is widely used as an industrial starter culture to conduct malolactic fermentation (MLF), the conversion of L‐malate to L‐lactate. Some cold‐climate wine regions still produce wines which prove too stressful for the application of a starter culture, resulting in sluggish or stuck MLFs which compromise the quality and stability of the wine. The literature on the three primary stress factors, low pH, ethanol and SO₂, is reviewed as well as their toxic mechanisms; possible interactions between the stress factors are also discussed. In particular, the toxic mechanisms of SO₂, an extremely inhibitory compound, are poorly understood. in vitro studies as well as in vivo studies in other prokaryotes point to numerous mechanisms which could have widespread impacts on the cell envelope, proteins and DNA in O. oeni. We also review the responses of O. oeni to these stress factors and offer a simplified model of events which cultures inoculated into particularly stressful wines may experience. Although significant advances have been made in our understanding of the stress responses in recent years, many fundamental aspects still require study.     

The effects of processing technology on the patulin content of juice during commercial apple juice concentrate production

 The effects of different treatments on the patulin content of apple juice during the production of industrial apple juice concentrate were investigated. Conventional clarification using a rotary vacuum precoat filter was found to be more effective than using ultrafiltration for the removal of patulin from apple juice. The average losses of patulin were 39% and 25% for conventional clarification and filtration, and ultrafiltration, respectively. Washing and handling appeared to be the most critical steps in removing patulin from apples since up to 54% could be removed using high-pressure water spraying. Received: 22 January 1998 / Revised version: 21 April 1998     

Standardisation of the detection of invert sugar addition to apple juice by capillary gas chromatography

 Capillary gas chromatography (CGC) can be used to check the authenticity of various fruit juices by analysing their oligosaccharide fingerprint profile. Thus, it enables the analyst to detect, for instance, the dilution of apple juice concentrates with cheaper sweeteners such as invert sugar syrup. In order to detect this, two marker peaks (IS1 and IS2) that are present in most invert sugar syrups are monitored in the CGC chromatogram of apple juice. In freshly pressed apple juice, these peaks are normally not found. As a result of apple juice processing and, in particular, the thermal treatment required to concentrate and pasteurise the juice, these peaks may however be present in authentic apple juice concentrate and juice made from it. It is nevertheless possible to distinguish between juice showing this "heat effect" and that to which invert sugar has been added since the IS2/IS1 peak ratio is very different in heated apple juice and in invert sugar syrups. Measuring this ratio and setting a maximum IS2/IS1 cut-off limit enables the analyst to assess whether the sample meets the set norms or has been adulterated. In the past, many laboratories experienced difficulties when comparing the results of this analysis and, in particular, IS2/IS1 peak ratios. We present evidence that this unsatisfactory situation is mainly due to the chromatographic behaviour of the IS1 peak. It appears indeed to be difficult to set up chromatographic conditions in all laboratories in order to produce reproducible results of the IS2/IS1 ratio. We have therefore examined the possibility of standardising results of this analysis and demonstrate that the comparison of any unknown apple juice sample with an external apple juice reference sample enables different laboratories to reach similar conclusions. The IS2/IS1 peak ratio in this reference sample was adjusted to the significant cut-off limit. The assessment (fail or pass) is thus based on a standardised IS2/IS1 ratio that is independent of several important analytical parameters which tend to strongly influence it. Received: 26 December 1997 / Revised version: 30 March 1998     

Standardisation of the detection of invert sugar addition to apple juice by capillary gas chromatography

 Capillary gas chromatography (CGC) can be used to check the authenticity of various fruit juices by analysing their oligosaccharide fingerprint profile. Thus, it enables the analyst to detect, for instance, the dilution of apple juice concentrates with cheaper sweeteners such as invert sugar syrup. In order to detect this, two marker peaks (IS1 and IS2) that are present in most invert sugar syrups are monitored in the CGC chromatogram of apple juice. In freshly pressed apple juice, these peaks are normally not found. As a result of apple juice processing and, in particular, the thermal treatment required to concentrate and pasteurise the juice, these peaks may however be present in authentic apple juice concentrate and juice made from it. It is nevertheless possible to distinguish between juice showing this "heat effect" and that to which invert sugar has been added since the IS2/IS1 peak ratio is very different in heated apple juice and in invert sugar syrups. Measuring this ratio and setting a maximum IS2/IS1 cut-off limit enables the analyst to assess whether the sample meets the set norms or has been adulterated. In the past, many laboratories experienced difficulties when comparing the results of this analysis and, in particular, IS2/IS1 peak ratios. We present evidence that this unsatisfactory situation is mainly due to the chromatographic behaviour of the IS1 peak. It appears indeed to be difficult to set up chromatographic conditions in all laboratories in order to produce reproducible results of the IS2/IS1 ratio. We have therefore examined the possibility of standardising results of this analysis and demonstrate that the comparison of any unknown apple juice sample with an external apple juice reference sample enables different laboratories to reach similar conclusions. The IS2/IS1 peak ratio in this reference sample was adjusted to the significant cut-off limit. The assessment (fail or pass) is thus based on a standardised IS2/IS1 ratio that is independent of several important analytical parameters which tend to strongly influence it. Received: 26 December 1997 / Revised version: 30 March 1998     

The effects of processing technology on the patulin content of juice during commercial apple juice concentrate production

 The effects of different treatments on the patulin content of apple juice during the production of industrial apple juice concentrate were investigated. Conventional clarification using a rotary vacuum precoat filter was found to be more effective than using ultrafiltration for the removal of patulin from apple juice. The average losses of patulin were 39% and 25% for conventional clarification and filtration, and ultrafiltration, respectively. Washing and handling appeared to be the most critical steps in removing patulin from apples since up to 54% could be removed using high-pressure water spraying. Received: 22 January 1998 / Revised version: 21 April 1998     

Investigating consumers' perception of apple juice as affected by novel and conventional processing technologies

This work evaluates consumers' perception of apple juice processed by high pressure processing (HPP) and pulsed electric field (PEF) compared with thermal processing. As a case study, young Chinese immigrants living in New Zealand were selected. Targeting a broad understanding of process impact, three industrially relevant apple cultivars (New Zealand Jazz, Rose and Granny Smith) were chosen. The consumer study was performed using napping with ultra‐flash profiling technique (n = 38). The process impact on sensory perception seems to vary among the investigated apple cultivars. For Jazz and Granny Smith apple cultivars, PEF‐ and HPP‐treated juices are perceived as fresh, natural, sweet and balanced flavour. For Rose apple cultivar, however PEF‐processed juices appear to be perceived as fresh flavour in comparison with HPP and thermally treated juices. Moreover, thermal processing caused cooked flavour. With respect to colour, immediately after processing, HPP retains the natural apple juice colour compared with other treatments.     

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。     

Influence of different pH values and inoculation time on the growth and malolactic activity of a strain of Oenococcus oeni

The present study was undertaken to evaluate the influence of medium pH and inoculation time on the growth and malolactic activity of an Oenococcus oeni culture. Samples of a commercial white grape juice adjusted to pH 3.2, 3.4 or 3.6, and inoculated with Saccharomyces cerevisiae strain CY3079, were inoculated with a malolactic culture ( Oenococcus oeni strain 31) at the beginning, middle, and end of alcoholic fermentation. The results obtained from this single case study show that it is possible to inoculate the bacterial culture at the three different times during alcoholic fermentation without slowing down or stopping alcoholic fermentation or causing failure of MLF. However, pH and timing of bacterial inoculation were critical to how rapidly MLF starts. At pH 3.2 a lowering of bacterial viability was observed, but a more important reduction was recorded at all tested pH levels when the bacteria were inoculated halfway through alcoholic fermentation. When inoculation was carried out at the end of alcoholic fermentation, the presence of yeast seemed to favour bacterial viability and activity and bacteria performed MLF even in difficult conditions such as pH values around 3. In all wines malolactic fermentation was accompanied by total degradation of malic and citric acids and production of L-lactic acid, D-lactic and acetic acids.     

Preliminary studies on microbial polysaccharides from different Penicillium species as flavour stabiliser in cloudy apple juice

This research deals with the production of water‐soluble polysaccharides from six polysaccharide‐producing Penicillium species. Eup. pinetorum ATCC 14770 was promising for exo‐ and endopolysaccharides production under various cultural conditions. pH 5.0, 7 days, and pH 8.0, 10 days, were the optimum for the production of exo‐ and endopolysaccharides, respectively, at 30 °C in medium containing glucose and peptone. HPLC results revealed that both polysaccharides were composed of pectin, cellulose and hemicellulose. Exopolysaccharides produced by Eup. pinetorum was subjected to evaluation as a natural flavour stabiliser in cloudy apple juice (CAJ). Twenty‐one volatile compounds classified into esters, aldehydes, alcohols and ketone were detected, only eighteen compounds of which were characterised by their odours threshold. The esters and alcohols are accounting for 87.64% and 5.44% of total volatiles, respectively. This study indicates that the use of exopolysaccharides as an alternative flavour stabiliser in CAJ is very promising.     

Analysis of the fouling mechanisms during cross-flow ultrafiltration of apple juice

The purpose of this work was to study the fouling mechanisms of a Carbosep^® M8 membrane during the cross-flow ultrafiltration of apple juice. A new fouling model has been developed that simultaneously considers membrane blocking within the pores, at the pore mouths and by cake formation at the membrane surface. Membrane fouling by apple juice was due to internal pore blocking as well as cake formation. When operating ultrafiltration at a transmembrane pressure of 150 kPa and a cross-flow velocity of 7 m/s, fouling was minimal with a gradual decrease of the relative contribution of cake formation; however, transmembrane pressure still exceeds critical pressure. The fouling model predicts no cake formation at a cross-flow velocity of 7.4 m/s and a transmembrane pressure of 150 kPa or at a cross-flow velocity of 7.0 m/s and a transmembrane pressure of 120 kPa. Under these conditions, internal membrane blocking would be the only mechanism responsible for the decrease of permeate flux.     

Inactivation of Escherichia coli by ozone treatment of apple juice at different pH levels

This research investigated the efficacy of gaseous ozone on the inactivation of Escherichia coli ATCC 25922 and NCTC 12900 strains in apple juice of a range of pH levels, using an ozone bubble column. The pH levels investigated were 3.0, 3.5, 4.0, 4.5 and 5.0. Apple juice inoculated with E. coli strains (10⁶ CFU/mL) was treated with ozone gas at a flow rate of 0.12 L/min and ozone concentration of 0.048 mg/min/mL for up to 18 min. Results show that inactivation kinetics of E. coli by ozone were affected by pH of the juice. The ozone treatment duration required for achieving a 5-log reduction was faster (4 min) at the lowest pH than at the highest pH (18 min) studied. The relationship between time required to achieve 5 log reduction (t_5d) and pH for both strains was described mathematically by two exponential equations. Ozone treatment appears to be an effective process for reducing bacteria in apple juice and the required applied treatment for producing a safe apple juice is dependant on its acidity level.     

Evaluation of the non-enzymatic browning in thermally processed apple juice by front-face fluorescence spectroscopy

Fresh apple juice was heated at 95 °C for 30 and 60 min, and then stored for 6 days for obtaining different extent of non-enzymatic browning. Front-face fluorescence excitation–emission-matrix (EEM) with excitation at 355 and 400 nm and emission ranges of 385–600, and 430–600 nm was used to measure the juice samples. The sign test pointed out an enhanced sensitivity of EEM compared to commonly used browning indicators, such as non-enzymatic browning index (NEBI), color readings in L^∗a^∗b^∗ space, and the concentration of 5-hydroxymethylfurfural (HMF). Using the fluorescence data, the correctness of classifications of fresh juice, heated juice, and stored juice were >85%. Multivariate analyses were applied to study the relations of fluorescence emission spectra and common indicators. For predicting NEBI, b^∗, and HMF, the correlation coefficients were >0.80. Resulting, the spectroscopic analyses of fluorescent neoformed products such as HMF provide a feasible approach to monitor non-enzymatic browning of juice.     

Application of multivariate statistical methods to extend the authenticity control of flavour constituents of apple juice

 The distribution of 15 aroma constituents, including the (R)- and (S)-enantiomers of ethyl 2-methylbutanoate, in 30 commercially available and laboratory prepared apple juices was evaluated by multivariate statistical methods. Principal component analysis and cluster analysis revealed ethyl 3-methylbutanoate, 3-methylbutyl acetate and (R)-ethyl 2-methylbutanoate as key components for the differentiation of apple juices. In freshly pressed and some commercial juices ethyl 3-methylbutanoate and 3-methylbutyl acetate were not present, and the (S):(R) ratio of ethyl 2-methylbutanoate was found to be >99 : 1%. Another group of commercial products was separated in which the amounts of ethyl 3-methylbutanoate and 3-methylbutyl acetate exceeded 1 μg/l and 180 μg/l, respectively, and in which the (S):(R) ratio of ethyl 2-methylbutanoate was <97 : 3%. The statistical data confirmed the high selectivity of apple fruit tissue in biosynthesizing 2-methylbutyl and 2-methylbutanoate esters, and could also be used to extend the results of enantio-multidimensional gas chromatography and other authenticity analyses. Received: 20 April 1998 / Revised version: 23 June 1998     

Effect of ozone processing on the colour,rheological properties and phenolic content of apple juice

Apple juice samples were ozonated with processing variables of ozone concentration (1–4.8% w/w) and processing time (0–10 min). Effects of processing variables on colour values (L, a and b), rheological properties and phenolic content were studied. Significant reductions in these parameters were observed during ozonation. Second order polynomial regression modelling was used to investigate the main effects of ozone concentration and processing time on the changes in the selected quality parameters of ozonated apple juice. Predicted models were found to be significant (p < 0.05) with low standard error and high coefficients of determination (R²).     

Choice probability for apple juice based on novel processing techniques: Investigating the choice relevance of mean-end-chains

The main aim of this study is to investigate an old challenge for means-end chain (MEC) theory; the lack of demonstration for means-end chains choice relevance. Even though MEC has become a commonly used tool in commercial market research and has been widely used in applied consumer research, within the core of academic consumer research, MEC has been almost ignored. One plausible explanation for this lack of interest may be that studies linking MEC data to choice have been few. In this study, we are to investigate how values and consequences generated from a previous MEC study structure can be linked to likelihood of choice.Hypotheses about European consumers’ likelihood of choice for novel processed juice are stated and tested in a rating based conjoint study in Norway, Denmark, Hungary and Slovakia. In the study, consumers probability of choice for high pressure processed (HPP) juice and pulsed electric field (PEF) juice are compared with their probability of choice for pasteurized juice and freshly produced apple juice, and consumer choices are tried explained by values and consequences generated from a MEC study. The study support, at least partly, that means-end chain structures’ have choice relevance.