Breeding of a high tyrosol‐producing sake yeast by isolation of an ethanol‐resistant mutant from a trp3 mutant

A novel breeding strategy for a high tyrosol‐producing sake yeast was developed by isolating an ethanol‐resistant mutant from a tryptophan auxotrophic mutant of a sake brewery yeast. Since tyrosol has antioxidant, cardioprotective and taste‐sharpening effects, increasing the tyrosol level of alcohol beverages could be beneficial in alcohol production. Since the transporters of aromatic amino acids are degraded by several stresses and mutants defective in the synthesis of aromatic amino acids are sensitive to ethanol, it was hypothesized that the degradation of these transporters should be inhibited in ethanol resistant mutants isolated from the auxotrophic mutants of aromatic amino acids, and that the uptake of aromatic amino acids would be increased in the mutants. Consistent with this hypothesis, sake was brewed with the ethanol‐resistant mutant of a tryptophan auxotrophic mutant and the sake was found to contain a lesser content of tyrosine and a higher content of tyrosol relative to the sake brewed with the parental strains. The taste of the sake brewed with the mutant strain could be discriminated from the sake brewed with the parental strains, probably because of the altered concentrations of tyrosol and certain amino acids and organic acids. The results suggest that combining the isolation of an ethanol‐resistant mutant and an auxotrophic mutant is an effective method to breed a brewing strain with a modified metabolism of these substances. Copyright © 2012 The Institute of Brewing & Distilling     

Sensitive detection of ochratoxin A in wine and cereals using fluorescence-based immunosensing

Ochratoxin A (OTA) is a mycotoxin found in a wide range of food and feedstuffs. Intake of OTA-contaminated food causes health concern due to the harmful effects reported on humans and animals. Much effort is currently devoted to set up and optimise highly sensitive and accurate methods of OTA analysis. This work describes the comparison of fluorescence-based immunosensing strategies for the analysis of OTA. First, an indirect competitive fluoroimmunoassay was designed and optimised. The assay enabled the quantification of the toxin at the levels set by the European legislation. Then, a flow-immunoassay based on kinetic exclusion measurements was developed. It showed the theoretical lowest limit of detection enabled by the affinity of the anti-OTA antibody (IC(80)=12ngL(-1) in the assay solution). Wine and cereal samples were analysed using the optimised flow system. No significant matrix effects were observed after simple pre-treatment of wine and OTA extraction from corn-flakes samples. This simple and highly sensitive automated biosensing-system allows OTA quantification in food and beverages. It is envisaged as a powerful tool for rapid and reliable toxin screening.     

Monitoring of acrylamide concentrations in potato chips in Japan between 2006 and 2010

Acrylamide levels in commercially available potato chips in Japan were monitored between August 2006 and June 2010 using the xanthydrol derivative gas chromatography-mass spectrometry (GC-MS) method. Seasonal and annual changes in acrylamide concentrations were determined. Nationwide bimonthly sampling of potato chips was carried out using a four-level design, and seasonal variations were detected in which the minimum acrylamide concentration was observed in August, and the maximum between February and June. Seasonal variations became less apparent after August 2008 as a result of annual effects and/or mitigation measures taken by the potato chip producers. Sampling uncertainties were separated into time-to-time, city-to-city, and lot-to-lot variation, and the largest variation was shown to be lot-to-lot including bag-to-bag.     

Deterministic regression methods for unbiased estimation of time-varying autoregressive parameters from noisy observations

A great deal of interest has been paid to autoregressive parameter estimation in the noise-free case or when the observation data are disturbed by random noise. Tracking time-varying autoregressive (TVAR) parameters has been also discussed, but few papers deal with this issue when there is an additive zero-mean white Gaussian measurement noise. In this paper, one considers deterministic regression methods (or evolutive methods) where the TVAR parameters are assumed to be weighted combinations of basis functions. However, the additive white measurement noise leads to a weight-estimation bias when standard least squares methods are used. Therefore, we propose two alternative blind off-line methods that allow both the variance of the additive noise and the weights to be estimated. The first one is based on the errors-in-variable issue whereas the second consists in viewing the estimation issue as a generalized eigenvalue problem. A comparative study with other existing methods confirms the effectiveness of the proposed methods.     

Metabolomics-based component profiling of hard and semi-hard natural cheeses with gas chromatography/time-of-flight-mass spectrometry, and its application to sensory predictive modeling

Gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS) was used to analyze hydrophilic low molecular weight components, including amino acids, fatty acids, amines, organic acids, and saccharides, in cheese, and the sensometric application for practical metabolomic studies in the food industry is described. Derivatization of target analytes was conducted prior to the GC/TOF-MS analysis. Data on 13 cheeses, six Cheddar cheeses, six Gouda cheeses and one Parmigiano-Reggiano cheese, were analyzed by multivariate analysis. The uniqueness of the Parmigiano-Reggiano cheese metabolome was revealed. Principal component analysis (PCA) showed no grouping of the Cheddar cheeses and Gouda cheeses according to production method or country of origin. The PCA loading plot confirms that many amino acids contribute positively to PC1, suggesting that PC1 is closely related to degradation of proteins, and that lactic acid contributed positively to PC2, whereas glycerol contributed negatively to PC2, suggesting that factors regarding degradation of carbohydrates and fats were expressed in PC2. Partial least squares (PLS) regression models were constructed to predict the relationship between the metabolite profile and two sensory attributes, "Rich flavor" and "Sour flavor", which were related to maturation. The compounds that play an important role in constructing each sensory prediction model were identified as 12 amino acids and lactose for "Rich flavor", and 4-aminobutyric acid, ornithine, succinic acid, lactic acid, proline and lactose for "Sour flavor". The present study revealed that metabolomics-based component profiling, focusing on hydrophilic low molecular weight components, was able to predict the sensory characteristics related to ripening.     

Enzymatic determination of acetoin in wines

An enzymatic micromethod for the determination of acetoin is described. The procedure involves spectrophotometric measurement of the amount of NADH utilised for the reduction of acetoin by 2,3-butanediol dehydrogenase at pH 6.5. The enzyme, partially purified from a strain of Sarcina, is quite stable at — 17 °C. Some other carbonyl compounds, especially acetaldehyde and diacetyl, interfere in the procedure. This interference can be eliminated by the addition of o-phenylenediamine to the sample solution. Acids, sugars and 2,3-butanediol do not interfere. By this method satisfactory results can be obtained in direct determination of acetoin in wines. Some red wines may require a decolorisation treatment before assay, but the treatment has little influence on the determination.     

A nondigestible saccharide,fructooligosaccharide, increases the promotive effect of a flavonoid, α‐glucosyl‐isoquercitrin,on glucagon‐like peptide 1 (GLP‐1) secretion in rat intestine and enteroendocrine cells

This study conducted in vivo and in situ experiments with rats to investigate the glucagon‐like peptide‐1 (GLP‐1) secretion in response to oral or ileal administration of α‐glucosyl‐isoquercitrin (20–40 mmol in 2 mL; Q3G), fructooligosaccharides (200 mmol in 2 mL; FOS) and Q3G + FOS. Direct effects on GLP‐1‐producing l ‐cells were also examined by an in vitro study using a murine enteroendocrine cell line, GLUTag. To evaluate the plasma GLP‐1 level, blood samples from jugular cannula for in vivo and portal cannula for in situ experiments were obtained before and after administration of Q3G, FOS, or Q3G + FOS. We found tendencies for increases but transient stimulation of GLP‐1 secretion by Q3G in in vivo and in situ experiments. Although FOS alone did not have any effects, Q3G + FOS enhanced and prolonged high plasma GLP‐1 level in both experiments. In addition, application of Q3G on GLUTag cells stimulated GLP‐1 secretion while FOS enhanced the effect of Q3G. Our results suggest that Q3G + FOS possess the potential for the management or prevention of Type 2 diabetes mellitus (T2DM) by enhancing and prolonging the GLP‐1 secretion via direct stimulation of GLP‐1 producing l ‐cell.     

Effect of “soft-electron” (low-energy electron) treatment on three stored-product insect pests

Developmental stages of three stored-product insect pests viz. Tribolium castaneum (Herbst), Plodia interpunctella (Hübner) and Callosobruchus chinensis (L.) were exposed to “soft electrons”, low-energy electrons. Soft electrons at an acceleration voltage of 170 kV effectively inactivated eggs, larvae and pupae of T. castaneum and P. interpunctella. The adults of T. castaneum and P. interpunctella were inactivated by treatment for 10 min (4.8 kGy) and 15 min (7.2 kGy), respectively. Soft electrons at 170 kV also inactivated the eggs of C. chinensis effectively. The adults of C. chinensis survived a 15 min exposure (7.2 kGy), but were inactivated having lost the ability to walk after a 5 min exposure (2.4 kGy). Soft electrons at 170 kV could not completely inactivate the larvae of C. chinensis inside beans, because the electrons with low penetration did not reach the larvae due to the shield of beans. The movement of T. castaneum larvae exposed to soft electrons was sluggish for several days, and then the larvae died with their hindguts emerged from their posterior ends 17 days after the treatment. DNA comet assay of cells of P. interpunctella larvae indicated that soft electrons at 170 kV damaged DNA in the larvae.