glutaminase and theanine

L-Glutaminase is considered to be an important industrial enzyme in both the pharmaceutical and food industries, especially for producing functional glutamyl compounds, such as l-theanine. Pseudomonas nitroreducens SP.001 with intracellular l-glutaminase activity has been screened previously. In the present study, three physical permeabilization methods were used to improve l-glutaminase activity. Then, the whole-cell immobilization conditions of permeabilized cells using sodium alginate as an embedding agent were optimized to enhance the enzyme's stability and reusability. The characteristics of the immobilized cells were investigated in comparison with those of permeabilized cells.. The results obtained showed that cell permeabilization using osmotic shock with 155 g L. The stability and reusability of the permeabilized cells were improved through the immobilization. These findings indicated that immobilized whole-cell l-glutaminase from P. nitroreducens SP.001 possesses more potential for various industrial biotechnological applications than free cells. © 2020 Society of Chemical Industry.

Topics: Alginates; Bacterial Proteins; Biocatalysis; Cells, Immobilized; Glutamates; Glutaminase; Pseudomonas

A flow-type microreactor containing glutaminase-mesoporous silica composites with 10.6 nm pore diameter (TMPS10.6) was developed for the continuous synthesis of theanine, a unique amino acid. High enzymatic activity was exhibited by the local control of the reaction temperature.

Topics: Biocatalysis; Glutamates; Glutaminase; Microfluidic Analytical Techniques; Particle Size; Porosity; Silicon Dioxide; Surface Properties

Theanine increases the antitumor effect of doxorubicin (DOX) with decreasing adverse reaction. We clarified the mechanism by which theanine decreases the adverse reaction of DOX on any metabolizing enzymes of theanine. There was no change in the activity of any CYPs and the cytochrome P450 content by theanine treatment. Namely, it was considered that the decrease of DOX adverse reactions by theanine was not connected with CYP activity. In other words, it is shown that theanine has no effect on the metabolism of other medicines and is safe as a food (tea) or supplement. Glutathione S-transferase activity did not change in the theanine-alone group whereas increased in the theanine and DOX-combined group. These results suggested that theanine combination increased the conjugate with DOX and GSH, promoted the efflux of GS-DOX conjugates from the liver, and decreased DOX concentration in the liver. In medium containing theanine with glutaminase in vitro, glutamate gradually generated, showing that glutaminase reacted with theanine. Furthermore, the generation of glutamate increased by reaction of theanine and gamma-glutamyltranspeptidase (gamma-GTP), showed that gamma-GTP converted theanine to glutamate. It is expected that theanine metabolism occurred by hydrolysis and rearrangement reaction by gamma-GTP in the liver. Namely, it is suggested that the metabolism of theanine mediated by glutaminase and gamma-GTP and the increase of glutamate mediated GSH is important for theanine-induced action. In conclusion, it appeared that theanine does not change the biodistribution of combined drugs but it modulates biodistribution or damage to the relative site of GSH, and shows preventive effects in tissue.

Topics: Animals; Antibiotics, Antineoplastic; Cytochromes; Cytosol; Doxorubicin; Glutamates; Glutaminase; Guanosine Triphosphate; Liver; Male; Mice; Microsomes, Liver; Mixed Function Oxygenases; Pharmaceutical Preparations

The distribution of theanine-degrading activity in Wistar rats was examined and this activity was detected only in the kidney. Judging from polyacrylamide gel electrophoresis, theanine-degrading enzyme from rat kidney was purified almost to homogeneity. Theanine-degrading activity was co-purified with glutaminase activity, and the relative activity for theanine was about 85% of that for L-glutamine throughout purification. Substrate specificity of purified enzyme preparation coincided well with the data of phosphate-independent glutaminase [EC 3.5.1.2], which had been previously reported. It was very curious that gamma-glutamyl methyl and ethyl esters were more effectively hydrolyzed than theanine and L-glutamine, in view of relative activity and K(m) value. It was suggested that gamma-glutamyl moiety in theanine molecule was transferred to form gamma-glutamylglycylglycine with relative ease in the presence of glycylglycine. On the other hand, purified phosphate-dependent glutaminase did not show theanine-degrading activity at all. Thus, it was concluded that theanine was hydrolyzed by phosphate-independent glutaminase in kidney and suggested that, as for the metabolic fate of theanine, its glutamyl moiety might be transferred by means of gamma-glutamyl transpeptidase reaction to other peptides in vivo.

Topics: Animals; Chromatography, Thin Layer; Electrophoresis, Polyacrylamide Gel; Ethylamines; gamma-Glutamyltransferase; Glutamates; Glutamic Acid; Glutaminase; Glycylglycine; Isoenzymes; Kidney; Phosphates; Rats; Rats, Wistar; Substrate Specificity