Odor thresholds of microbially induced off-flavor compounds in apple juice

Microbially derived off-flavor is a major problem in apple juice production as it diminishes the sensory quality of the juice significantly. Fifteen relevant off-flavor compounds that are formed in apple juice, for example, by the strains Alicyclobacillus acidoterrestris and Actinomycetes (Streptomyces ssp.) were investigated with respect to their sensory relevance. The odor threshold values (i.e., detection and recognition values) were determined for all compounds in the matrix apple juice. Odor threshold values for fenchyl alcohol are reported here for the first time. The obtained values were set in relation to the limits of detection and quantification of a previously published GC-MS method. Eight tainted apple juice samples were analyzed for the presence of the 2 strains and the 15 off-flavor compounds. Both strains could be found in the samples; the presence of Streptomyces ssp. as spoilage bacteria of apple juice is reported for the first time. In samples with distinct off-flavor, 2-isopropyl-3-methoxypyrazine, 2-isobutyl-3-methoxypyrazine, 2-methylisoborneol, 1-octen-3-ol, fenchyl alcohol, geosmin, and guaiacol as well as 2,6-dibromophenol were determined in concentrations higher than the detection threshold.     

Mousy off-flavor: a review

Although mousy off-flavor occurs infrequently in wine, it can be economically disastrous to the wine producer as, at worst, it can render the wine unpalatable or, at best, decrease the quality of the wine resulting in a lower sale price. Wines infected with either lactic acid bacteria (LAB) (particularly heterofermentative strains) or Dekkera/Brettanomyces yeast can potentially produce mousy off-flavor. There are three known compounds that cause mousy off-flavor: 2-ethyltetrahydropyridine, 2-acetyltetrahydopyridine, and 2-acetylpyrroline. Dekkera/Brettanomyces have been shown to be capable of producing at least two of these compounds, whereas LAB are capable of producing all three. The reason as to why mousy off-flavor forms in some wines and not in others is still not fully understood. The issue is further complicated by the fact that the compounds that have thus far been identified as necessary for off-flavor formation are all potentially available in wine (e.g., ethanol, L-lysine, L-ornithine, and metal ions). For these reasons, the microbe's metabolism probably plays a key role in mousy off-flavor formation. In the case of Dekkera/Brettanomyces-induced mousy off-flavor, it appears that oxygen may play a key role. Thus, a wine infected with Dekkera/Brettanomyces in the absence of oxygen may not become mousy unless exposed to oxygen via a processing or handling procedure.     

Enzymatic removal of off-flavors from apple juice

Contaminating microorganisms such as Actinomycetes, Alicyclobacillus, and Chlostridium can generate off-flavors in apple juices. Such bacterial metabolites represent, besides phenol types such as guaiacol and 2,6-dibromophenol, a broad range of other chemicals, for example, geosmin, 2-methylisoborneol, or alpha-terpineol. A laccase from Trametes hirsuta was purified, immobilized, and applied for the selective elimination of off-flavor substances in apple juice caused by microbial contamination. The evaluation using GC-MS showed that enzymatic treatment could reduce the amount of guaiacol and 2,6-dibromophenol in apple juice significantly by 99 and 52%, respectively. Upon addition of mediators, the degradation could be increased and the spectrum of substrates extented. Furthermore, commercial apple juices spiked with off-flavors were treated in a continuous-flow reactor and tested by sensory evaluation.     

Identification of the medicinal off-flavor compound formed from ascorbic acid and (E)-hex-2-enal

A test apple beverage made up of apple juice (20%), high-fructose corn syrup (11.5%), citric acid (0.43%), trisodium citrate (0.02%), apple-odor flavor (0.1%), and ascorbic acid (0.02%) was stored at 40 °C and then analyzed for the change of odor in the beverage. Although no thermoacidophilic bacteria (TAB) were detected, a medicinal off-flavor was perceived after the 8 weeks of storage. Model experiments on the ingredients of the test apple beverage revealed that the off-flavor compound had been formed by ascorbic acid and (E)-hex-2-enal. Synthesis and NMR (1H, 13C, HMQC, and HMBC) analyses identified the compound as 6-propylbenzofuran-7-ol. The odor quality, retention index (RI), and mass spectrum of synthetic 6-propylbenzofuran-7-ol were identical with those of the medicinal odor compound from the test apple beverage. Sensory evaluation revealed the recognition thresholds for medicinal odor were 31.4 ppb in water and 24.0 ppb in apple beverage, and the detection thresholds were 19.6 ppb in water and 8.6 ppb in apple beverage, respectively. The quantified concentration of 6-propylbenzofuran-7-ol formed in test apple beverage was 90 ppb, approximately. This concentration was well above the odor threshold, so it was concluded that the compound was the source of the medicinal off-flavor.     

Light-induced off-flavor development in cloudy apple juice

Cloudy apple juice has been found to develop off-flavors during storage in daylight. The development of off-flavors and volatile compounds was monitored in reconstituted juice prepared from 'Golden Delicious' and 'Fuji' apple concentrates stored in glass bottles under fluorescent light (3000 lx, 8 degrees C). A strong metallic off-flavor was formed by photooxidation. A major contributor to the off-flavor was identified as 1-octen-3-one by gas chromatography-olfactometry. In addition, six volatile compounds, pentanal, 2-methyl-1-penten-3-one, hexanal, (E)-2-heptenal, 6-methyl-5-hepten-2-one, and (E)-2-octenal, increased significantly after light exposure and could contribute to the off-flavor. Except for pentanal and hexanal, these volatiles were found only after light exposure. Higher levels of volatiles were observed in juice from 'Golden Delicious' apples than in juice from 'Fuji' apples, and this difference was consistent with higher levels of suspended solids. When the suspended solids were removed by centrifugation, the development of volatiles on exposure to light was reduced significantly.     

Isolation and characterization of lactic acid bacteria from feces of newborn baby and from dongchimi

Lactic acid bacteria were screened from feces of newborn baby and from dongchimi. Selection criteria employed included the ability of strains to withstand environmental conditions such as low pH, high bile concentration, and oxygen. The isolates were applied to the juice of various vegetables, and fermentabilities of isolates were compared. Strains F20-3, F35-3, and F35-6 showed high stability compared to the other strains at pH 3.0 and 2.3. Strains D1 and D2 showed the highest survival at pH 3.0 and survived at 1% high bile concentration. The selected strains were able to survive at low pH and relatively high bile concentration and were not affected by oxygen. The growth of isolates was >10(7) cfu/mL in natural media, and strains were not affected by the pH values of the vegetables. Therefore, isolated strains are thought to survive through the intestinal ecosystem and are considered to be suitable for application of the fermented product using various vegetables for their functionality. The isolates were identified as Lactobacillus plantarum and Lactobacillus fermentum.     

Fermentation of plant cell wall derived polysaccharides and their corresponding oligosaccharides by intestinal bacteria

New types of nondigestible oligosaccharides were produced from plant cell wall polysaccharides, and the fermentation of these oligosaccharides and their parental polysaccharides by relevant individual intestinal species of bacteria was studied. Oligosaccharides were produced from soy arabinogalactan, sugar beet arabinan, wheat flour arabinoxylan, polygalacturonan, and rhamnogalacturonan fraction from apple. All of the tested substrates were fermented to some extent by one or more of the individual species of bacteria tested. Bacteroides spp. are able to utilize plant cell wall derived oligosaccharides besides their reported activity toward plant polysaccharides. Bifidobacterium spp. are also able to utilize the rather complex plant cell wall derived oligosaccharides in addition to the bifidogenic fructooligosaccharides. Clostridium spp., Klebsiella spp., and Escherichia coli fermented some of the selected substrates in vitro. These studies do not allow prediction of the fermentation in vivo but give valuable information on the fermentative capability of the tested intestinal strains.     

苹果树腐烂病拮抗细菌鉴定及其抑菌作用效果测定

为了开发一种高效低毒的苹果树腐烂病生防制剂,通过对峙培养法、形态学及分子生物学的方法进行了苹果树腐烂病菌拮抗菌的分离筛选及鉴定,采用离体枝条法测定了拮抗菌对苹果树腐烂病的防效,并采用显微观察和液体培养法分别研究了拮抗菌对苹果树腐烂病菌的抑菌机理和无菌滤液对苹果树腐烂病菌生长的影响。分离筛选结果表明,从甘肃省各苹果产区果园土壤和苹果树枝条上分离得到23株细菌,2株对苹果树腐烂病菌具有较好拮抗作用,分别为LZ-1201和TS-1203,其对苹果树腐烂病菌菌丝生长抑制率分别为79.00%和85.00%。鉴定结果表明,菌株LZ-1201和TS-1203分别为枯草芽孢杆菌(Bacillus subtilis)和解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。离体枝条防效测定表明,拮抗菌无菌滤液对苹果树腐烂病的防效随着稀释倍数增大而降低,原液防效最高,分别为74.43%和77.07%。抑菌作用机理结果表明,两株拮抗菌均可导致苹果树腐烂病菌丝膨大畸形、原生质浓缩、释放及溶解。拮抗菌无菌滤液对腐烂病菌生长的影响测定结果表明,无菌滤液对腐烂病菌分生孢子萌发和菌丝生长量均有显著抑制作用(P0.05),其无菌滤液稀释40倍时对腐烂病菌分生孢子萌发和菌丝生长量的抑制率均高于60%,表明该拮抗菌具有很好的生防潜力。     

苹果树根际高效解钾菌的筛选及鉴定

为探明苹果树根际解钾菌的种群特征和解钾性能,从苹果树根际土壤中分离获得解钾能力较强的高效解钾菌。本研究以钾长石为唯一钾源,分离获得118株具有解钾活性的菌株,重复片段PCR基因指纹分析(Repetitive-element PCR,rep-PCR)聚为29个类群。利用火焰分光光度法测定菌株解钾能力,获得5株高效解钾菌,平均解钾活性达到41.47mg·L-1,其中K105的解钾能力最强,有效态钾增长23.09%。采用H2O2消煮后测得的K+浓度升高,有效态钾增长最高达31.22%。采用形态特征观察、生理生化特性检测和基于16S r RNA基因序列的系统发育分析对高效解钾菌株进行鉴定。结果表明:K1为不动杆菌属(Acinetobacter sp.)、K98、K105和K115为假单胞菌属(Pseudomonas sp.)、K168为芽孢杆菌属(Bacillus sp.)。研究结果可为开辟新型高效解钾菌,为连作土壤的改良和苹果专用微生物菌肥的开发提供依据。     

Development of liquid chromatography-electrospray mass spectrometry for the determination of patulin in apple juice: investigation of its contamination levels in Japan

Patulin is a mycotoxin produced by mainly Penicillium species, for example, P. expansum, and Aspergillus species. There are several reports of patulin contamination in apple juice. Last year, the Ministry of Health, Labour and Welfare of Japan set the maximum allowable level of patulin in apple juice at 50 ppb and decided that the measurement of patulin levels in apple juice products should be conducted. To this end, a simple, accurate, and selective analytical method for the detection of patulin at levels lower than 5 ppb, the detection limit, is desired. This paper reports the development of an analytical method that employs solid-phase extraction-liquid chromatography-mass spectrometry (SPE-LC-MS). When MS measurements were conducted with the selected ion monitoring (SIM) mode, the pseudomolecular ions at m/z 153 and 156 were used to monitor patulin and (13)C(3)-labeled patulin, respectively. The detection limit (S/N = 3) and the quantification limit (S/N = 10) of patulin at injection levels into LC-MS were 12.5 and 25 pg, respectively. However, when the actual sample was applied for the analysis based on the developed method including the sample preparation, the detection limit (S/N = 3) and quantification limit (S/N = 10) were 2.5 and 5 pg in sample, respectively. The calibration curve obtained for concentrations ranging from 5 to 500 ppb showed good linearity with a coefficient of determination (r (2)) of 0.999. In addition, the recovery was >95% when an internal standard was used. The method was applied to the analysis of 76 apple juice samples from Japan, and as a result, patulin levels ranging from <1.0 to 45 ppb (detection frequency = 15/76) were detected. In this study, it was found that patulin was a greater contaminant in concentration/reduction than in "not from concentrate" apple juice.     

Red grape juice inhibits iron availability: application of an in vitro digestion/caco-2 cell model

Adequate bioavailable Fe intake is essential for optimal growth and intellectual development of infants and children. Fruit juices are nutritious and popular drinks for infants and children and are known to contain Fe uptake inhibitors (e.g., polyphenolic compounds) and a dominant promoter, ascorbic acid. Ascorbic acid is naturally present in fruit juices and is added during processing to almost all juices found in supermarkets. With these facts taken into account, an in vitro digestion/Caco-2 cell culture model was developed to compare the effects of apple, pear, white grape, red grape, prune, grapefruir, and orange juices on iron bioavailability. In two series of experiments, juices from a local supermarket were combined with FeCl(3) or commercial infant cereal fortified with elemental iron and subject to simulated gastric and intestinal digestion. Caco-2 cell ferritin formation in response to exposure to the digests served as the measure of Fe uptake. The pear, apple, grapefruit, orange, and white grape juice significantly increased Fe bioavailability from FeCl(3). For the infant cereal studies, the apple, orange, pear, and white grape juices increased the Fe bioavailability of the infant cereal. In contrast, the red grape juice and prune juice had profound inhibitory effects on iron bioavailability. These inhibitory effects were likely due to high levels of polyphenolic compounds that bind and thereby prevent absorption of soluble Fe. These inhibitory compounds appeared to counteract the promotional effects of ascorbic acid as they were in considerable molar excess relative to ascorbic acid and Fe in the digest. From a nutritional standpoint, the results suggest that individuals in need of optimal Fe absorption should avoid red grape and prune juice or at least vary the types of juices consumed. Alternatively, individuals seeking to limit Fe uptake (e.g., hemochromatitics and astronauts) may be able to utilize red grape or prune juice as effective inhibitors of Fe uptake. Consumers should be aware that the compounds that inhibit Fe availability are also linked to anticancer benefits; thus, a dietary balance of the above juices may be optimal.     

Investigation of different apple cultivars by high resolution magic angle spinning NMR. A feasibility study

(1)H HR-MAS NMR spectroscopy was applied to apple tissue samples deriving from 3 different cultivars. The NMR data were statistically evaluated by analysis of variance (ANOVA), principal component analysis (PCA), and partial least-squares-discriminant analysis (PLS-DA). The intra-apple variability of the compounds was found to be significantly lower than the inter-apple variability within one cultivar. A clear separation of the three different apple cultivars could be obtained by multivariate analysis. Direct comparison of the NMR spectra obtained from apple tissue (with HR-MAS) and juice (with liquid-state HR NMR) showed distinct differences in some metabolites, which are probably due to changes induced by juice preparation. This preliminary study demonstrates the feasibility of (1)H HR-MAS NMR in combination with multivariate analysis as a tool for future chemometric studies applied to intact fruit tissues, e.g. for investigating compositional changes due to physiological disorders, specific growth or storage conditions.     

gamma-Radiation influences browning, antioxidant activity, and malondialdehyde level of apple juice

Apple juice was gamma-irradiated at 5 degrees C at doses ranging from 0 to 8.9 kGy and then stored at 5 degrees C for 15 days. Ionizing radiation reduced the browning of apple juice and increased antioxidant activity measured by the ferric-reducing antioxidant power (FRAP) assay. The magnitude of changes increased with radiation dose. The level of malondialdehyde (MDA) measured using the thiobarbituric acid reactive substrates assay increased at radiation doses above 2.67 kGy. The browning of irradiated juices increased during storage at 5 degrees C, but the irradiated juices were still lighter than controls at the end of storage. Differences in FRAP values disappeared during early periods of storage while higher MDA levels were observed in irradiated samples during most of the storage period. Elimination of suspended matter from apple juice did not alter irradiation-induced changes in browning, FRAP, or MDA formation. As compared to irradiation conducted at 5 and 20 degrees C, treatment at -15 degrees C was less effective in reducing browning and in increasing MDA formation but elevated FRAP values. The exclusion of oxygen from juices did not affect the reduction in browning due to irradiation but promoted the increase in FRAP values and decreased the irradiation-induced MDA formation.     

Polyphenols are intensively metabolized in the human gastrointestinal tract after apple juice consumption

Polyphenols are secondary plant compounds showing anticarcinogenic effects both in vitro and in animal experiments and may thus reduce the risk of colorectal cancer in man. The identification of polyphenol metabolites formed via their passage through the small intestine of healthy ileostomy subjects after apple juice consumption is presented. Identification and quantification of polyphenols and their metabolites were performed using HPLC-DAD as well as HPLC-ESI-MS/MS. Total procyanidin content (TPA) was measured, and additionally the mean degree of polymerization (DPm) of the procyanidins was determined in the apple juice and ileostomy effluents. As products of polyphenol metabolism, D-(-)-quinic acid and methyl esters of caffeic acid and p-coumaric acid are liberated from the corresponding hydroxycinnamic acid esters. 1-Caffeoylquinic acid and 3-caffeoylquinic acid were determined as products of isomerization. Phloretin 2'-O-glucoside (phloridzin) and phloretin 2'-O-xyloglucoside were metabolized into the corresponding aglycons phloretin and phloretin 2'-O-glucuronide and all were found in the ileostomy effluent. Ninety percent of the consumed procyanidins were recovered in the ileostomy effluent and therefore would reach the colon under physiologic circumstances. The DP m was reduced (DP m of apple juice=5.7) and varied depending on the time point of excretion. The gastrointestinal passage seems to play an important role in the colonic availability of apple polyphenols.     

Reduction of azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl residues in processed apples

McIntosh, Red Delicious, and Golden Delicious from two years of experimental spray programs using azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl were processed into frozen apple slices, applesauce, single-strength juice, and juice concentrate. Residue levels were expressed as micrograms per 150 g of apple or the equivalent amount of apple product to calculate the percentage change in these pesticides brought about by processing. Producing single-strength apple juice reduced azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl residues by 97.6, 100, 97.8, and 78.1%, respectively. Production of applesauce reduced all four compounds by >/=95%. Azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl residues were reduced in apple slices by 94.1, 85.7, 98.6, and 94.7%, respectively. Processing is shown to be very effective in reducing the levels of these pesticides.     

Activity and concentration of polyphenolic antioxidants in apple juice. 3. Stability during storage

Kinetic data are reported describing the stability of various classes of polyphenolic antioxidants in an apple juice enriched in these compounds as a function of storage temperature and oxygen concentration. The most thermally sensitive compounds were the various quercetin glycosides and epicatechin, whereas phloridzin and chlorogenic acid were more stable. The quercetin glycosides showed differences in their stability: quercetin galactoside approximately quercetin rhamnoside > quercetin glucoside/rutinoside > quercetin arabinoside. The effect of the presence of oxygen on the degradation rates was clear for only quercetin and to a lesser extent for epicatechin. Accelerated shelf-life testing of enriched apple juice during 4 days at 80 degrees C showed decreases in the antioxidant activity of 20-40%. The parameters obtained were used to predict the stability at different storage conditions. Calculations showed that polyphenolic antioxidants and antioxidant activity of enriched apple juice will be quite stable at ambient or refrigerated storage conditions up to half a year.     

A significant proportion of indigenous rhizobia from India associated with soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) distinctly belong to <Emphasis Type="Italic">Bradyrhizobium</Emphasis> and <Emphasis Type="Italic">Ensifer</Emphasis> genera

The diversity among 269 rhizobia isolated from naturally occurring root nodules of soybean collected from two different agro-ecological regions of India, based on RFLP and sequences of the intergenic spacer (IGS) between the 16S and 23S rRNA genes, growth rate, and indole acetic acid production, revealed their significant, site-dependent genomic diversity. Among these bacteria, nine IGS genotypes were identified with two endonucleases. They were distributed into five divergent lineages by sequence analysis of each IGS representative strain, i.e., (1) comprising IGS genotypes I, II, III, and reference Bradyrhizobium yuanmingense; (2) with genotype IV and strains of unclassified bradyrhizobia genomic species; (3) including genotypes V, VI, and Bradyrhizobium liaoningense; (4) with IGS genotype VII and Bradyrhizobium elkanii strains; and (5) comprising IGS genotypes VIII, IX, and different Ensifer genus bacteria. Host-specificity test revealed that all rhizobia-nodulated soybean and cowpea and only part of them formed nodules on Arachis hypogeae and Cajanus cajan. The great diversity of soybean nodulators observed in this study emphasises that Indian soil is an important reservoir of nitrogen-fixing rhizobia.     

苹果汁冷冻浓缩工艺的研究

应用渐进冷冻浓缩原理对苹果汁冷冻浓缩特性进行研究,试验证明渐进浓缩苹果汁浓缩效果良好。影响冷冻浓缩的主要因素是搅拌速度。搅拌速度1200 r/min时浓缩效果最佳,溶质损失最少。冷冻浓缩对苹果汁中酸度和维生素C含量无影响,是提高浓缩品质量的较佳方法。浓缩产品感官质量均匀一致,保持了果汁的原有风味。     

Characterization of haze-active proteins in apple juice

The nature of the haze-active protein (HAP) in apple juice was investigated. Heat treatment removed protein indiscriminately while polyvinylpolypyrrolidone (PVPP) treatment was fairly specific for proteins of 15 and 28 kDa. Presumably, the PVPP bound to polyphenols, which in turn were complexed with protein. Three candidate apple HAPs were isolated. Two were extracted from juice with acetone and fractionated by hydrophobic interaction chromatography and solid phase extraction with C18 (HAP I) or SAX (HAP II) material. Hydroxyproline-rich protein was extracted from apple tissue (HAP III). The order of haze formation with tannic acid was gliadin > HAP III > HAP II > HAP I > bovine serum albumin, which shows increasing haze formation with increasing proline content. The sizes of HAP I, II, and III were 28, 15, and 12 kDa; the first two corresponded to the sizes of proteins removed by PVPP treatment and are involved in juice haze formation.