A novel bionic in vitro bioelectronic tongue based on cardiomyocytes and microelectrode array for bitter and umami detection.

Electronic tongues (ETs) have been developed and widely used in food, beverage and pharmaceutical fields, but limited in sensitivity and specificity. In recent years, bioelectronic tongues (BioETs) integrating biological materials and various types of transducers are proposed to bridge the gap between ET system and biological taste. In this work, a bionic in vitro cell-based BioET is developed for bitter and umami detection, utilizing rat cardiomyocytes as a primary taste sensing element and microelectrode arrays (MEAs) as a secondary transducer for the first time. The primary cardiomyocytes of Sprague Dawley (SD) rats, which endogenously express bitter and umami taste receptors, were cultured on MEAs. Cells attached and grew well on the sensor surface, and syncytium was formed for potential conduction and mechanical beating, indicating the good biocompatibility of surface coating. The specificity of this BioET was verified by testing different tastants and bitter compounds. The results show that the BioET responds to bitter and umami compounds specifically among five basic tastants. For bitter recognition, only those can activate receptors in cardiomyocytes can be recognized by the BioET, and different bitter substances could be discriminated by principal component analysis (PCA). Moreover, the specific detections of two bitters (Denatonium Benzoate, Diphenidol) and an umami compound (Monosodium Glutamate) were realized with a detection limit of 10-6 M. The cardiomyocytes-based BioET proposed in this work provides a new approach for the construction of BioETs and has promising applications in taste detection and pharmaceutical study.

Isolation and identification of the umami enhancing compounds in Japanese soy sauce.

To clarify the key compounds that account for the umami taste of soy sauce, a typical Japanese soy sauce, Koikuchi Shoyu, was separated by preparative chromatography, and the umami enhancing fractions were screened on the basis of an umami intensity of a 6.0 mM monosodium L-glutamate (MSG) solution. Liquid chromatography-time of flight mass spectrometry (LC-TOFMS), 1D/2D nuclear magnetic resonance spectroscopy (NMR) studies of the umami enhancing fractions led to the identification of N-(1-deoxy-D-fructos-1-yl)pyroglutamic acid (Fru-pGlu), N-(1-deoxy-D-fructos-1-yl)valine (Fru-Val), N-(1-deoxy-D-fructos-1-yl)methionine (Fru-Met), pyroglutamylglutamine (pGlu-Gln), and pyroglutamylglycine (pGlu-Gly). Although all the compounds identified were at sub-threshold concentrations in the soy sauce, a taste reconstitution experiment revealed that they contributed part of the umami taste of the soy sauce.

History of glutamate production.

In 1907 Kikunae Ikeda, a professor at the Tokyo Imperial University, began his research to identify the umami component in kelp. Within a year, he had succeeded in isolating, purifying, and identifying the principal component of umami and quickly obtained a production patent. In 1909 Saburosuke Suzuki, an entrepreneur, and Ikeda began the industrial production of monosodium l-glutamate (MSG). The first industrial production process was an extraction method in which vegetable proteins were treated with hydrochloric acid to disrupt peptide bonds. l-Glutamic acid hydrochloride was then isolated from this material and purified as MSG. Initial production of MSG was limited because of the technical drawbacks of this method. Better methods did not emerge until the 1950s. One of these was direct chemical synthesis, which was used from 1962 to 1973. In this procedure, acrylonitrile was the starting material, and optical resolution of dl-glutamic acid was achieved by preferential crystallization. In 1956 a direct fermentation method to produce glutamate was introduced. The advantages of the fermentation method (eg, reduction of production costs and environmental load) were large enough to cause all glutamate manufacturers to shift to fermentation. Today, total world production of MSG by fermentation is estimated to be 2 million tons/y (2 billion kg/y). However, future production growth will likely require further innovation.

Intramolecular carbon isotopic composition of monosodium glutamate: biochemical pathways and product source identification.

Monosodium glutamate (MSG) obtained as trade samples from several manufacturers was studied to determine the range of its intramolecular 13C/12C composition. Although the carbon isotopic composition of the total MSG molecule did not differ among manufacturers in most instances, significant differences were observed in the isotopic composition of the alpha-carboxyl carbon, suggesting that different proprietary strains of industrial microorganisms or MSG purification methods may impart unique isotopic fingerprints upon their products. The 13C depletion of the alpha-carboxyl carbon relative to the rest of the molecule helps constrain the identity of the potential anapleurotic carboxylating enzymes responsible for its fixation.

Study on the column process of adsorption decolorization for monosodium glutamate solution in fluidized beds.

The adsorption decolorization of monosodium glutamate solution employed in multiple column fluidized beds in a series is studied. The equilibrium data, the mass transfer kinetics, and fluid flow parameters are determined. A mathematics model taking into account the effects of particle size classification, particle size distribution, external and internal diffusions, and axial mixing in the liquid and solid phases is proposed, and the simulated results are in good agreement with the experimental data.

Improved cleanup and derivatization for gas chromatographic determination of monosodium glutamate in foods.

A gas chromatographic procedure is described for determining monosodium glutamate (MSG) in several types of food. A sample is extracted with acetone-water (1 + 1). Acetone is evaporated and an aliquot of the extract is buffered with 1M NH4OH-1M NH4Cl pH 9 solution, and chromatographed directly on a column of QAE Sephadex A-25 that has been pretreated with the same buffer. MSG is eluted with 0.1N HCl, and a portion of the eluate is evaporated to dryness and reacted with dimethylformamide(DMF)-dimethylacetal to form the glutamic acid derivative, which is injected into a gas chromatograph and measured by flame ionization detection. Recoveries of MSG from sample fortified at 5-500 mg ranged from 92.8 to 100%.