Glutamate Solution Research Articles

Saltiness Enhancement by the Characteristic Flavor of Dried Bonito Stock

There is a pressing need for the development of ways of preparing palatable salt-reduced foods to reduce the salt intake of the Japanese population. The salt-reducing effect of the characteristic flavors other than umami of dried bonito stock, which is widely used in everyday Japanese food, was examined by sensory evaluation. In the 1st sensory evaluation, the effect was evaluated in a model solution. The saltiness of 0.80% NaCl solution was equivalent to that of 0.12% monosodium glutamate (MSG) solution containing 0.81% NaCl and dried bonito stock containing 0.68% NaCl. Saltiness enhancement could not be found when MSG solution was used, but was found with 6% dried bonito stock. The 2nd evaluation examined whether the effect was valid for 2 everyday Japanese foods--traditional Japanese clear soup (sumashi-jiru) and steamed egg custard (tamagodoufu). Although enhancement of saltiness by dried bonito stock could not be clearly demonstrated in the soup, a change in NaCl concentration within 15% did not affect the palatability of the soup. However, dried bonito stock not only enhanced the saltiness but also improved the palatability of steamed egg custard. These findings are expected to be useful for improving the palatability of salt-reduced food.

Mercaptoacetate inhibition of fatty acid β-oxidation attenuates the oral acceptance of fat in BALB/c mice

We investigated the effect of beta-oxidation inhibition on the fat ingestive behavior of BALB/c mice. Intraperitoneal administration to mice of mercaptoacetate, an inhibitor of fatty acid oxidation, significantly suppressed intake of corn oil but not intake of sucrose solution or laboratory chow. To further examine the effect of mercaptoacetate on the acceptability of corn oil in the oral cavity, we examined short-term licking behavior. Mercaptoacetate significantly and specifically decreased the number of licks of corn oil within a 60-s period but did not affect those of a sucrose solution, a monosodium glutamate solution, or mineral oil. In contrast, the administration of 2-deoxyglucose, an inhibitor of glucose metabolism, did not affect the intake or short-term licking counts of any of the tasted solutions. These findings suggest that fat metabolism is involved in the mechanism underlying the oral acceptance of fat as an energy source.

Kinetics and mechanisms of depolymerization of alginate and chitosan in aqueous solution

The kinetics and mechanisms of depolymerization of aqueous chitosan and alginate solutions at elevated temperatures have been investigated. Chitosan salts of different degree of acetylation (FA), type of counterions (-glutamate, -chloride) and degree of purity were studied. One commercially available highly purified sodium alginate sample with high content of guluronic acid (G) was also studied. Furthermore, the influence of oxygen, H+ and OH− ions on the initial depolymerization rates was investigated. Depolymerization kinetics was followed by measuring the time courses of the apparent viscosity and the intrinsic viscosity. The initial rate constants for depolymerization were determined from the intrinsic viscosity data converted to a quantity proportional to the fraction of bonds broken. The activation energies of the chitosan chloride and chitosan glutamate solutions with pH close to 5 and the same degree of acetylation, FA=0.14, were determined from the initial rate constants to be 76±13kJ/mol and 80±11kJ/mol, respectively.The results reported herein suggest that the stability of aqueous chitosan and alginate solutions at pH values 5–8 will be influenced by oxidative–reductive depolymerization (ORD) as the primary mechanism as long as transition metal ions are presented in the samples. Acid – and alkaline depolymerization will be the primary mechanisms for highly purified samples.

Isoflurane Activates Human Cardiac Mitochondrial Adenosine Triphosphate-Sensitive K+ Channels Reconstituted in Lipid Bilayers

Activation of the mitochondrial adenosine triphosphate (ATP)-sensitive K+ channel (mitoK(ATP)) has been proposed as a critical step in myocardial protection by isoflurane-induced preconditioning in humans and animals. Recent evidence suggests that reactive oxygen species (ROS) may mediate isoflurane-mediated myocardial protection. In this study, we examined the direct effect of isoflurane and ROS on human cardiac mitoK(ATP) channels reconstituted into the lipid bilayers. Inner mitochondrial membranes were isolated from explanted human left ventricles not suitable for heart transplantation and fused into lipid bilayers in symmetrical potassium glutamate solution (150 mM). ATP-sensitive K+ currents were recorded before and after exposure to isoflurane and H2O2 under voltage clamp. The human mitoK(ATP) was identified by its sensitivity to inhibition by ATP and 5-hydroxydecanoate. Addition of isoflurane (0.8 mM) increased the open probability of the mitoK(ATP) channels, either in the presence or absence of ATP inhibition (0.5 mM). The isoflurane-mediated increase in K+ currents was completely inhibited by 5-hydroxydecanoate. Similarly, H2O2 (200 microM) was able to activate the mitoK(ATP) previously inhibited by ATP. These data confirm that isoflurane, as well as ROS, directly activates reconstituted human cardiac mitoK(ATP) channel in vitro, without apparent involvement of cytosolic protein kinases, as commonly proposed. Activation of the mitoK(ATP) channel may contribute to the myocardial protective effect of isoflurane in the human heart.

A cell-physiological description of GE, a GTP-binding protein that mediates exocytosis.

Introduction of GTP gamma S or other non-metabolic analogues of GTP into permeabilized myeloid granulocytes (mast cells, eosinophils, neutrophils) constitutes a sufficient stimulus to induce exocytosis. We concentrate on mast cells. Exocytosis from cells permeabilized in isotonic glutamate solution proceeds in the absence of ATP and at exceedingly low levels (< 10(-9) M) of Ca2+. Mg2+ strongly promotes GTP gamma S-induced exocytosis but this requirement can be spared and then obliterated by lifting Ca2+ through 10(-7) to 10(-6) M. GTP provides only a modest support to exocytosis but becomes almost equipotent with GTP gamma S when Mg2+ is excluded. Ca2+ alone is unable to induce exocytosis. We envisage that the terminal stage of exocytosis (membrane fusion) requires activation of GE, a putative GTPase so far undefined as a molecular entity. Ca2+, presumed to act through a Ca(2+)-binding protein (CE, also undefined) supports exocytosis by promoting the exchange of guanine nucleotides on GE. In the absence of Mg2+ the onset of exocytosis is characterized by delays that have concentration-dependent (binding) and independent components. The latter are sensitive to the identity of the stimulating nucleotide (GTP < GTP gamma S < Gpp [NH]p) and may reflect activation of GE. The activation by Ca2+ and Mg2+ and the delays preceding onset of GTP gamma S-triggered exocytosis are reminiscent of the action of glucagon and Mg2+ in the activation of adenylate cyclase in hepatocyte membranes. The cell-physiological description predicts GE to be an alpha beta gamma heterotrimeric GTP-binding protein with functional similarity to GS.

The Maxi-Chloride Channel in Human Syncytiotrophoblast: A Pathway for Taurine Efflux in Placental Volume Regulation?

Taurine (Tau), the most abundant amino acid in fetal blood, is highly concentrated in human placenta. During pregnancy, Tau is involved in the neurological development of the fetus, and in volume regulation of the placenta. The placenta may release taurine in parallel with K + and Cl − in response to an increase in cell volume. However, the pathway for the volume-activated taurine efflux is unknown. One candidate is a voltage-dependent Maxi-chloride channel from apical syncytiotrophoblast membrane (MVM), with a conductance over 200 pS and multiple subconductance states. Our aim was to study whether this channel could be a Tau conductive pathway in the MVM. Purified human placental MVM were reconstituted into giant liposomes suitable for patch clamp recordings. Typical Maxi-chloride channel activity was detected in symmetrical chloride (Cl −) solutions, and then taurine (Tau), Aspartate (Asp), and glutamate (Glu) solutions were used in the bath of excised patches to detect single channel currents carried by these anions. The relative permeabilities (P), estimated from the shift in reversal potential of current-voltage curves after anion replacement, were as follows: Chloride > Taurine = Glutamate = Aspartate. In Tau symmetric conditions using equivalent Cl − concentrations, the slope conductance was 62.4 ± 7.3 pS. The data shows that Tau and other amino acids diffuse through the Maxi-chloride channel, which could be of great importance as part of the mechanism involved in the volume regulation process in human placenta.

Endogenous Erythropoietin Signaling Is Required for Normal Neural Progenitor Cell Proliferation

Erythropoietin (Epo) and its receptor (EpoR), critical for erythropoiesis, are expressed in the nervous system. Prior to death in utero because of severe anemia EpoR-null mice have fewer neural progenitor cells, and differentiated neurons are markedly sensitive to hypoxia, suggesting that during development Epo stimulates neural cell proliferation and prevents neuron apoptosis by promoting oxygen delivery to brain or by direct interaction with neural cells. Here we present evidence that neural progenitor cells express EpoR at higher levels compared with mature neurons; that Epo stimulates proliferation of embryonic neural progenitor cells; and that endogenous Epo contributes to neural progenitor cell proliferation and maintenance. EpoR-null mice were rescued with selective EpoR expression driven by the endogenous EpoR promoter in hematopoietic tissue but not in brain. Although these mice exhibited normal hematopoiesis and erythrocyte production and survived to adulthood, neural cell proliferation and viability were affected. Embryonic brain exhibited increased neural cell apoptosis, and neural cell proliferation was reduced in the adult hippocampus and subventricular zone. Neural cells from these animals were more sensitive to hypoxia/glutamate neurotoxicity than normal neurons in culture and in vivo. These observations demonstrate that endogenous Epo/EpoR signaling promotes cell survival in embryonic brain and contributes to neural cell proliferation in adult brain in regions associated with neurogenesis. Therefore, Epo exerts extra-hematopoietic function and contributes directly to brain development, maintenance, and repair by promoting cell survival and proliferation independent of insult, injury, or ischemia.

Molecular and Sensory Characterization of γ-Glutamyl Peptides as Key Contributors to the Kokumi Taste of Edible Beans (Phaseolus vulgaris L.)

Addition of a nearly tasteless aqueous extract isolated from beans (Phaseolus vulgaris L.) to a model chicken broth enhanced its mouthfulness and complexity and induced a much more long-lasting savory taste sensation on the tongue. Gel permeation chromatography and hydrophilic interaction liquid chromatography/comparative taste dilution analysis (HILIC/cTDA), followed by LC-MS/MS and 1D/2D-NMR experiments, led to the identification of gamma-L-glutamyl-L-leucine, gamma-L-glutamyl-L-valine, and gamma-L-glutamyl-L-cysteinyl-beta-alanine as key molecules inducing this taste-modifying effect. Sensory analysis of aqueous solutions of these peptides showed threshold concentrations between 3.3 and 9.4 mmol/L for an unspecific, slightly astringent sensation. More interestingly, when added to a savory matrix such as sodium chloride and monosodium glutamate solutions or chicken broth, the detection thresholds of these gamma-glutamyl peptides decreased significantly and remarkably enhanced mouthfulness, complexity, and long-lastingness of the savory taste were observed; for example, the threshold of gamma-glutamyl-cysteinyl-beta-alanine decreased by a factor of 32 in a binary mixture of glutamic acid and sodium chloride. As tasteless molecules inducing mouthfulness, thickness, and increasing continuity of savory foods were coined about 10 years ago as "kokumi" flavor compounds, the peptides identified in raw as well as thermally treated beans have to be considered as kokumi compounds.

Transverse relaxation in rat optic nerve

In vitro investigations were performed to study the proton T(2) relaxation spectrum of rat optic nerve. Studies in which the nerve was incubated in a D(2)O-based solution revealed that >98% of the spectrum originated from the water protons. The spectrum was found to consist of three components having relaxation times and sizes similar to those reported in the literature for peripheral nerve. Procedures were taken to confirm the existence of a third optic nerve component and that it was not an artifact of the long-lived water protons of the in vitro incubation solution. Evidence using paramagnetic agents in the incubation solution, which removed the two longest-lived nerve components from the spectrum, revealed the existence of a small fourth component (<10% of total) having a T(2) relaxation time similar to that of the intermediate-lived nerve component. Bathing the nerves in a 10 mM glutamate solution, glutamate known to result in cellular swelling in mammalian central nervous system (CNS), was found to increase the component size of the longest-lived nerve component, suggestive that this component may result from cellular water.

Effect of glutamate ingestion on whole-body glutamate turnover in healthy elderly and patients with chronic obstructive pulmonary disease

Objective Decreased whole-body glutamate turnover is found in healthy elderly and in patients with chronic obstructive pulmonary disease (COPD). Glutamate supplementation as an option to increase whole-body glutamate turnover and, hence, glutamate availability has never been investigated. In the present study, we developed a protocol based on repeated glutamate ingestion to increase plasma glutamate concentration to a steady-state level without inducing toxic side effects and to evaluate the effect of repeated glutamate ingestion on whole-body glutamate turnover in patients with COPD and healthy elderly. Methods In part 1, the response of plasma glutamate concentration was determined in young healthy volunteers who repeatedly ingested a glutamate solution. The tolerance of the glutamate drink was evaluated in 26 healthy volunteers by a food tolerance questionnaire. In part 2, eight male patients with COPD and eight healthy elderly ingested the glutamate drink, an isomolar amount of a glutamine drink, or only water to test the effect on plasma glutamate concentration and whole-body glutamate turnover. Results In part 1, repeated ingestion of 30 mg of glutamate per kilogram of body weight every 20 min increased plasma glutamate concentration five-fold to steady-state level within 80 min and without any side effects. In part 2, repeated ingestion of glutamate significantly increased whole-body glutamate turnover in healthy controls and patients with COPD, although the increase was smaller in patients with COPD than in controls. Conclusion We found that repeated ingestion of 30 mg of glutamate per kilogram of body weight every 20 min can increase glutamate availability in healthy elderly and patients with COPD, who are likely more dependent on external glutamate ingestion than are young adults.

Precipitation of L‐Glutamic Acid: Determination of Nucleation Kinetics

AbstractThis work presents a new procedure to determine the nucleation kinetics during the reactive precipitation of an organic substance. The new method has been applied to the pH‐shift precipitation of L‐glutamic acid upon mixing of an aqueous monosodium glutamate solution with hydrochloric acid. The induction time has been measured precisely in a stirred batch reactor by the combination of two in‐situ measurement techniques, namely ATR‐FTIR spectroscopy (attenuated total reflection Fourier Transform technique) and Focused Beam Reflectance Measurement (FBRM). It is shown that ATR‐FTIR is a suitable tool to measure the concentrations of the different L‐glutamic acid ions, and can be used to determine the starting time of the process when the desired supersaturation is established in the reactor. The onset of particle formation is detected through FBRM. The precipitated polymorph is identified using in‐situ Raman spectroscopy and Particle Vision &amp; Measurement (PVM). Finally, the induction time is used together with the independently measured growth kinetics to determine the nucleation rate.

Real-time monitoring of L-glutamate release from mouse brain slices under ischemia with a glass capillary-based enzyme electrode

Real-time monitoring of L-glutamate release from various neuronal regions of mouse hippocampal slices under ischemia (a glucose-free hypoxia condition) is described. A glass capillary microelectrode with a tip size of approximately 10 microm containing a very small volume ( approximately 2 microL) of a solution of glutamate oxidase (GluOx) and ascorbate oxidase was used. First, the amperometric response behavior of the electrode at 0 V versus Ag/AgCl was characterized with a standard glutamate solution in terms of continuous measurements, effect of oxygen, viscosity of solution and concentration dependence. The electrode was applied to the real-time monitoring of L-glutamate released from different neuronal regions of acute hippocampal slices submerged in a hypoxia solution. The time-resolved amounts of L-glutamate released at various neuronal regions (CA1, CA3 and DG) of mouse hippocampal slices were quantified and compared with the reported L-glutamate fluxes using difference-image analysis during ischemia.

Effect of electroacupuncture on metabolism of lipids in rats of obesity induced by sodium glutamate (Abstract)

To explore mechanisms of acupuncture for reducing of body weight. The rat obesity model was developed by subcutaneous injection of sodium glutamate solution, and the experimental rats were divided into a blank control group, a model group, an electroacupuncture group and a sibutramine group; blood TC, TG, HDL-C, LDL-C contents and lipoprotein lipase (LPL) activity, and serum leptin and insulin levels were determined. The body weight and Lee's index, and TG, TC and LDL-C contents in the electroacupuncture group decreased significantly as compared with those in the model group (all P<0.01), the actions in the electroacupuncture group being superior to those in the sibutramine group; the HDL-C contents in both the electroacupuncture and the sibutramine groups were higher than that of the model group (P<0.01) with no significant difference between the electroacupuncture group and the sibutramine group; LPL activity in the electroacupuncture group increased significantly as compared with that in the model group (P<0.01); serum leptin and insulin levels in both the electroacupuncture group and the sibutramine group were lower than that in the model group (P<0.05, P<0.01), with no significant difference between the electroacupuncture group and the sibutramine group in the effect on insulin content, and the decrease of leptin level in the electroacupuncture group was more as compared with the sibutramine group (P<0.01). Electroacupuncture can improve the status of high blood lipids, increase lipoprotein lipase activity, and regulate serum leptin and insulin levels in the obestic rat.

Monitoring of intercellular messengers released from neuron networks cultured in a microchip

A cellular biochemistry analysis system was integrated on a quartz glass microchip with a microchamber for cell culture followed by a microchannel for detecting with a thermal lens microscope (TLM). Nerve cells from rat hippocampus were successfully cultured to form neural networks in the microchip. An aqueous solution of glutamate, which is known as a neurotransmitter, was introduced to stimulate the cultured neuron to release a retrograde messenger, arachidonate which is considered to be critical for neuronal plasticity, especially for long-term potentiation (LTP). After the introduction, the solution that flowed through the culture chamber was analyzed using the UV-TLM (excitation wavelength, 244 nm). The measured signal intensity was dependent on glutamate solution concentration, and the neurons were considered to release the retrograde messenger according to the glutamate concentration. This system is suitable for time-course monitoring of ultra trace amounts of chemicals released from very small amount of cultured cells.

Two Regions in the N-terminal Domain of Ionotropic Glutamate Receptor 3 Form the Subunit Oligomerization Interfaces That Control Subtype-specific Receptor Assembly

The N-terminal domain (NTD) of alpha-amino-3-hydroxy-5-methylisoxazolepropionate (AMPA) and kainate glutamate receptors plays an important role in controlling subtype specific receptor assembly. To identify NTD subdomains involved in this process we generated AMPA glutamate receptor 3 (GluR3) mutants having intra-NTD substitutions with the corresponding regions of the kainate receptor GluR6 and tested their ability to form functional heteromers with wild-type subunits. The chimeric design was based on the homology of the NTD to the NTD of the metabotropic GluR1, shown to form two globular lobes and to assemble in dimers. Accordingly, the NTD was divided into four regions, termed here N1-N4, of which N1 and N3 correspond to the regions forming lobe-1 and N2 and N4 to those forming lobe-2. Substituting N1 or N3 impaired functional heteromerization but allowed protein-protein interactions. Conversely, exchanging N2 or N4 preserved functional heteromerization, although it significantly decreased homomeric activity, indicating a role in subunit folding. Moreover, a deletion in GluR3 corresponding to the hotfoot mouse mutation of the glutamate receptor delta2, covering part of N2, N3, and N4, impaired both homomeric and heteromeric oligomerization, thus explaining the null-like mouse phenotype. Finally, computer modeling suggested that the dimer interface, largely formed by N1, is highly hydrophobic in GluR3, whereas in GluR6 it contains electrostatic interactions, hence offering an explanation for the subtype assembly specificity conferred by this region. N3, however, is positioned perpendicular to the dimer interface and therefore may be involved in secondary interactions between dimers in the assembled tetrameric receptor.

Analysis of Purified Tubulin in High Concentration of Glutamate for Application in High Throughput Screening for Microtubule-Stabilizing Agents

High-throughput screening (HTS) is a powerful approach for the discovery of potential and effective drugs from a number of candidates. The dynamics of tubulin assembly and disassembly in vitro are an important target for the screening of anti-tumor agents. However, previously described methods are not amenable for HTS. In this paper, we compared preparation methods of tubulin and suggest a combination method, i.e., one cycle of assembly and disassembly following ion-exchange chromatography using high concentrations of glutamate to increase the recovery of tubulin with high purity. The resultant highly purified microtubule-associated proteins-free tubulin in high glutamate solution was directly employed in tubulin polymerization assays. Our results indicate that the system is feasible and practicable as a model for HTS for microtubule-stabilizing agents.

Characterisation of weak magnetic field effects in an aqueous glutamic acid solution by nonlinear dielectric spectroscopy and voltammetry

BackgroundPrevious reports indicate altered metabolism and enzyme kinetics for various organisms, as well as changes of neuronal functions and behaviour of higher animals, when they were exposed to specific combinations of weak static and alternating low frequency electromagnetic fields. Field strengths and frequencies, as well as properties of involved ions were related by a linear equation, known as the formula of ion cyclotron resonance (ICR, abbreviation mentioned first by Liboff). Under certain conditions already a aqueous solution of the amino acid and neurotransmitter glutamate shows this effect.MethodsAn aqueous solution of glutamate was exposed to a combination of a static magnetic field of 40 μT and a sinusoidal electromagnetic magnetic field (EMF) with variable frequency (2–7 Hz) and an amplitude of 50 nT. The electric conductivity and dielectric properties of the solution were investigated by voltammetric techniques in combination with non linear dielectric spectroscopy (NLDS), which allow the examination of the dielectric properties of macromolecules and molecular aggregates in water. The experiments target to elucidate the biological relevance of the observed EMF effect on molecular level.ResultsAn ion cyclotron resonance (ICR) effect of glutamate previously reported by the Fesenko laboratory 1998 could be confirmed. Frequency resolution of the sample currents was possible by NLDS techniques. The spectrum peaks when the conditions for ion cyclotron resonance (ICR) of glutamate are matched. Furthermore, the NLDS spectra are different under ICR- and non-ICR conditions: NLDS measurements with rising control voltages from 100–1100 mV show different courses of the intensities of the low order harmonics, which could possibly indicate "intensity windows". Furthermore, the observed magnetic field effects are pH dependent with a narrow optimum around pH 2.85.ConclusionsData will be discussed in the context with recent published models for the interaction of weak EMF with biological matter including ICR. A medical and health relevant aspect of such sensitive effects might be given insofar, because electromagnetic conditions for it occur at many occasions in our electromagnetic all day environment, concerning ion involvement of different biochemical pathways.

Mild hypothermia inhibits exogenous glutamate-induced increases in nitric oxide synthesis.

The purpose of this study was to investigate changes in nitric oxide (NO) synthesis induced by exogenous glutamate perfusion into the cerebral cortex, and the effects of mild hypothermia on this glutamate-induced NO synthesis. Glutamate-induced cortical lesions were produced by perfusion of 0.5 M glutamate solution via a microdialysis probe, and the extracellular concentrations of NO end-products (nitrite and nitrate) were measured by microdialysis in normothermic (37 degrees C) and hypothermic (32 degrees C) rats. The levels of NO end-products in the normothermia group were elevated markedly by glutamate perfusion, and this change was completely attenuated by the induction of hypothermia. The glutamate-induced increases were also attenuated markedly by both Nomega-nitro-L-arginine methyl ester (L-NAME) and 7-nitroindazole (7-NI). These results suggest that the perfusion of exogenous glutamate into the cortex induces NO synthesis, that is derived primarily from the activity of neuronal NO synthase. These results also demonstrate that hypothermia prevents this glutamate-induced increase in NO, suggesting that the protection afforded by the hypothermic condition is most likely linked to its inhibition of the glutamate-induced NO synthesis.

Sensitivity of particle sizing by ultrasonic attenuation spectroscopy to material properties

Ultrasonic attenuation spectroscopy is a recently developed technique for rapid characterisation of particulate suspensions at high concentrations. Implementation of the full Epstein–Carhart–Allegra–Hawley model, which is used to transform ultrasound attenuation measurements into particle size and concentration information, requires knowledge of seven physical properties of the particulate phase and of a further seven properties of the continuous phase. Reliable data are not always available for all these properties. In this study, an assessment is made of the influence of inaccuracy in the physical properties on the recovered values of particle size and concentration. Two systems of organic crystals, glutamic acid crystals in aqueous solution of glutamic acid and monosodium glutamate crystals in aqueous solution of monosodium glutamate, are investigated over sizes from 1 to 100 μm. It is found that the same properties are significant for both material systems generally. These are the densities of both phases, shear rigidity of the particles, and sound speed and attenuation of the continuous phase. For these material systems, the size and concentration returned by analysis of ultrasound attenuation are insensitive to several properties which are required for the full Epstein–Carhart–Allegra–Hawley model: thermal dilation, thermal conductivity and heat capacity of both phases, and sound attenuation of the particulate phase.

A combined solid-state and submerged cultivation integrated with adsorptive product extraction for production of Monascus red pigments.

A process taking advantages of combined solid-state and submerged cultivation of Monascus for red pigment production and integration of a product removal unit was developed. The solid-state cultivation was carried out in a 5 l fermentor, with rice being used not only as the substrate but also the support for Monascus. The inclusion of rice submergence and integration of product separation were achieved by intermittently rinsing the rice with monosodium glutamate (MSG) solutions every 12 h followed by an adsorptive extraction of the red pigment dissolved in the rinsing solution. With this new process, the Monascus red pigment production was increased by 24% as compared with that by the plain fixed-bed cultivation.