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Properties of glutamic acid decarboxylase from edible mushroom
Takahiro Yoshida,Takao Terashita,Norifumi Shirasaka 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4
[Object] Gamma-Amino Butyric Acid (GABA), a four-carbon non protein amino acid, is widely distributed in nature and acts as the major suppressive neurotransmitter in the mammalian central nervous system. Recently, GABA has been reported to have several physiological functions such as antihypertensive, diuretic, relaxing and antidiabetic effects. Due to these unique biological functions, GABA has been used as functional ingredients for functional foods. Glutamic Acid Decarboxylase (GAD), which catalyzes decarboxylation of L-glutamic acid to GABA, has been purified from mammals, higher plants, and some microorganisms and their properties have already been reported. On the other hand, mushrooms are commonly appreciated as healthy food due to their nutritional properties, such as low calorie, rich in fiber, vitamin and mineral. L-Glutamic acid and GABA are also commonly distributed in edible mushrooms. The fact that mushrooms accumulate GABA suggests the existence of GAD. However, the enzyme property of mushroom GAD has not been reported. In the present study, we tried to evaluate the property of Flammulina veltipes GAD, and the purification and characterization of enzyme is also investigated. [Methods and Results] Mushroom (fruit-body of Flammulina veltipes) used in this study was purchased in local market. GAD activity was determined by formation of GABA from L-glutamic acid in the presence of pyridoxal-5’- phosphate (PLP). The activity of enzyme was determined semi-quantitatively by color intensity of GABA on TLC analysis. Fruit-body was crushed and then centrifuged to separate supernatant and precipitate. To investigate the location of GAD, both supernatant and precipitate were subjected to enzyme reaction after dialysis. The enzyme activity of precipitate is stronger than that of supernatant. The formation of GABA was observed between pH 4 and 6 and the maximum color intensity of GABA was observed at pH 6. However, GAD activity was lost after dialysis for overnight against buffer of pH 6-11. These results suggest that GAD from Flammulina veltipes is stable at pH 4-5 in spite of its optimum pH for GABA production is around 6.
Yoshida Haruno,Kim Jung-Min,Maeda Takahiro,Goto Mieko,Tsuyuki Yuzo,Shibata Sachiko,Shizuno Kenichi,Okuzumi Katsuko,Kim Jae-Seok,Takahashi Takashi 대한진단검사의학회 2023 Annals of Laboratory Medicine Vol.43 No.3
Background: Comparative analysis of virulence factors (VFs) between Pasteurella canis and Pasteurella multocida are lacking, although both cause zoonotic infections. We determined the virulence-associated genome sequence characteristics of P. canis and assessed the toxin gene prevalence unique to P. canis among clinical isolates of P. canis and P. multocida. Methods: We selected 10 P. canis and 16 P. multocida whole-genome sequences (WGSs) from the National Center for Biotechnology database. The VFanalyzer tool was used to estimate P. canis-characteristic VFs. Amino acid sequences of VFs were compared with multiple-aligned sequences. The genome structure containing P. canis-characteristic and adjacent loci was compared to the corresponding P. multocida genome structure. After designing primer sequences and assessing their accuracy, we examined the gene prevalence of the P. canis-characteristic VFs using PCR among clinical isolates of P. multocida and P. canis. Results: Using VFanalyzer, we found virulence-associated cytolethal distending toxin (cdt)A–cdtB–cdtC loci common to all P. canis WGSs that were not found in P. multocida WGSs. Similarities in the multiple alignments of CdtA–CdtB–CdtC amino acid sequences were found among the 10 P. canis WGSs. Shared or similar loci around cdtA–cdtB–cdtC were identified between the P. canis and P. multocida genome structures. The PCR-based cdtA–cdtB–cdtC prevalence differed for P. canis and P. multocida clinical isolates. Conclusions: P. canis-specific cdtA–cdtB–cdtC prevalence was identified among clinical isolates. These three loci may be unique toxin genes and promising targets for the rapid identification of P. canis in clinical settings.
Mechanical Analysis of Pion Capture Superconducting Solenoid System for COMET Experiment at J-PARC
Iio, Masami,Yoshida, Makoto,Ye Yang,Taekyung Ki,Okamura, Takahiro,Sasaki, Ken-ichi,Makida, Yasuhiro,Ogitsu, Toru,Mihara, Satoshi,Nakamoto, Tatsushi,Sugano, Michinaka,Terashima, Akio,Kawamata, Hiroshi IEEE 2017 IEEE transactions on applied superconductivity Vol.27 No.4
<P>KEK is currently constructing a long series of superconducting solenoid beamline for the COMET Phase-I experiment at the Hadron facility of J-PARC. The magnet system consists of a 5-T pion capture solenoid, a curved muon transport solenoid, and a large bore detector solenoid. The pion capture solenoid system consists of a large cold mass including four coils with an inner diameter of 1340 mm and a small cold mass including six small coils with an inner diameter of 500 mm. The peak magnetic field on the conductor reaches to 5.4 T at an operation current of 2700 A. This paper presents the design and structural details of a magnet, radiation-resistant coil, cold mass and vacuum vessel, apart from the results of the electromagnetic analysis in two-dimensional (2-D) and 3-D.</P>