Chemo-enzymatic hybrid process for production of monatin, a high intensity sweetener

Monatin, 4-hydroxy-4-(3-indolylmethyl)-glutamic acid, is a naturally occurring sweet amino acid isolated from the plant Sclerochiton ilicifolius, found in South Africa [1]. Monatin has two asymmetric centers at C2 and C4, the (2R,4R)-monatin isomer has been found to be the sweetest among its four stereoisomers. It is 2700 times sweeter than sugar and has a clean taste like sugar. Because of these properties, (2R,4R)-monatin has been expected as an new high-intensity sweetener [2]. However, industrial production process of (2R,4R)-monatin using inexpensive raw materials has not been established owing to the difficulty for optically specific synthesis. Here, we report a chemo-enzymatic hybrid process for production of (2R,4R)-monatin from l-tryptophan. In the steps of enzymatic reaction from l-tryptophan, l-amino acid deaminase and aldolase were used for production of 4-(Indole-3-ylmethyl)-4-hydroxy-2-oxoglutaric acid (IHOG) with pyruvic acid as co-substrate. The keto-form of (2R,4R)-monatin, (R)-IHOG, was specifically synthesized by using R-specific aldolase from Shingomonas sp. in the second reaction. In the next chemical reaction steps, (R)-IHOG was converted to the oxime form, reduced to (2R,4R) and (2S,4R)-monatin, and (2R,4R)-monatin salt was obtained from optical resolution by crystallization. By the combination of epimerization and crystallization, (2R,4R)-monatin was obtained specifically from the mixture of diastereomers. In this study, we established an efficient production process for (2R,4R)-monatin using both chemical and enzymatic reactions, and a large amount of (2R,4R)-monatin was prepared by the bench-scale production. Please click Additional Files below to see the full abstract

鉛直下向き揚力を発生する没水翼の造波現象に関する研究

研究期間:平成6-7年度 ; 研究種目:一般研究B ; 課題番号: 0645235

Radioimmunoassay of Plasma 17α-Hydroxyprogesterone

A radioimmunoassay for the measurement of 17α-hydroxyprogesterone (17α-OHP) in plasma was investigated utilizing antiserum produced by the introduction of 17α-hydroxyproges- terone-3-oxim BSA. Accuracy was such that within-assay variance was 16.2% and between-assay variance 18.3%. The 17α-OHP mean plasma levels were 930±201 pg/ml(n=8)for normal adult males and 402±186 pg/ml(n=11)in the follicular phase and 1,190±662 pg/ml(n=12)in the luteal phase of females with a normal menstrual cycle. The 17α-OHP levels in some tissues and tissue fluid also were measured, being 18.06±12.91 ng/ml(n=6) in the normal trophoblastic tissue, 1.7±0.5 ng/ml(n=17) in the trophoblastic tissue of a hydatidiform mole and 855.5±507.4 ng/ml(n=5)in lutein cyst fluid

Radioimmunoassay of Estradiolusing Estradiol-6-oxim-BSA Anti Serum

A radioimmunoassay (RIA) of estradiol was performed with the use of antiserum produced by estradiol-6-oxim-BSA. The precision of the assay was such as to result in a within-assay variance of 10.1% and a between-assay variance of 31.8%. The concentrations of estradiol thus determined in the serum of women with normal menstrual cycles were 69.0±40.6 pg/ml inthe menstrual phase, 127.8±70.3 pg/ml in the follicular phase, 281.9±113.5 pg/ml in the ovulatory phase, and 165.4±79.5 pg/ml in the luteal phase. The blood estradiol level durinh pregnancy ranged from 0.28 to 13.17 ng/ml in the first trimester, 2.6 to 48.1 ng/ml in the second trimester, and 13.5 to 130.0 nh/ml in the third trimester

Elevated expression of CD30 in adult T-cell leukemia cell lines: possible role in constitutive NF-κB activation

BACKGROUND: Human T-cell leukemia virus type 1 (HTLV-1) is associated with the development of adult T-cell leukemia (ATL). HTLV-1 encoded Tax1 oncoprotein activates the transcription of genes involved in cell growth and anti-apoptosis through the NF-κB pathway, and is thought to play a critical role in the pathogenesis of ATL. While Tax1 expression is usually lost or minimal in ATL cells, these cells still show high constitutive NF-κB activity, indicating that genetic or epigenetic changes in ATL cells induce activation independent of Tax1. The aim of this study was to identify the molecules responsible for the constitutive activation of NF-κB in ATL cells using a retroviral functional cloning strategy. RESULTS: Using enhanced green fluorescent protein (EGFP) expression and blasticidin-resistance as selection markers, several retroviral cDNA clones exhibiting constitutive NF-κB activity in Rat-1 cells, including full-length CD30, were obtained from an ATL cell line. Exogenous stable expression of CD30 in Rat-1 cells constitutively activated NF-κB. Elevated expression of CD30 was identified in all ATL lines examined, and primary ATL cells from a small number of patients (8 out of 66 cases). CONCLUSION: Elevated CD30 expression is considered one of the causes of constitutive NF-κB activation in ATL cells, and may be involved in ATL development

Dual-Matrix Domain-Wall: A Novel Technique for Generating Permutations by QUBO and Ising Models with Quadratic Sizes

The Ising model is defined by an objective function using a quadratic formula of qubit variables. The problem of an Ising model aims to determine the qubit values of the variables that minimize the objective function, and many optimization problems can be reduced to this problem. In this paper, we focus on optimization problems related to permutations, where the goal is to find the optimal permutation out of the $n!$ possible permutations of $n$ elements. To represent these problems as Ising models, a commonly employed approach is to use a kernel that utilizes one-hot encoding to find any one of the $n!$ permutations as the optimal solution. However, this kernel contains a large number of quadratic terms and high absolute coefficient values. The main contribution of this paper is the introduction of a novel permutation encoding technique called dual-matrix domain-wall, which significantly reduces the number of quadratic terms and the maximum absolute coefficient values in the kernel. Surprisingly, our dual-matrix domain-wall encoding reduces the quadratic term count and maximum absolute coefficient values from $n^3-n^2$ and $2n-4$ to $6n^2-12n+4$ and $2$, respectively. We also demonstrate the applicability of our encoding technique to partial permutations and Quadratic Unconstrained Binary Optimization (QUBO) models. Furthermore, we discuss a family of permutation problems that can be efficiently implemented using Ising/QUBO models with our dual-matrix domain-wall encoding.Comment: 26 pages, 9 figure

Acoustic stiffness and change in plug cartilage over time after autologous osteochondral grafting: correlation between ultrasound signal intensity and histological score in a rabbit model

We investigated quantitative changes over time in ultrasound signal intensity (an index of stiffness), signal duration (an index of surface irregularity), and interval between signals (an index of thickness) of plug cartilage in an animal model of autologous osteochondral grafting. A full-thickness osteochondral plug was surgically removed and replaced in male Japanese white rabbits (n = 22). Specimens obtained at day 0 and weeks 2, 4, 8, 12 and 24 postoperatively were assessed using an ultrasound system and by macroscopic and histological evaluation (modified Mankin's score). Histology revealed that the plug sank until 2 weeks postoperatively, and that newly formed cartilage-like tissue covered the plug, but at 24 weeks the tissue detached. The plug itself survived well throughout the period of observation. Although the signal intensity at the plug site was same as that in the sham operated contralateral knee at day 0, from 2 to 24 weeks postoperatively it was less than that in the sham knee. At 8 weeks, this difference was significant (P < 0.05). Modified Mankin's score revealed early degenerative changes at the site, but macroscopic examination did not. Signal intensity correlated significantly with score (both at day 0 and at the five postoperative time points [P < 0.05, r = -0.91] and as a whole [P < 0.05, r = -0.36]). Signal intensity also significantly correlated with the individual subscores for 'cartilage structure' (P < 0.05, r = -0.32) and 'cartilage cells' (P < 0.05, r = -0.30) from the modified Mankin's score, but not significantly with subscores for 'staining' and 'tidemark'. Signal duration correlated significantly with total score (as a whole [P < 0.05, r = 0.34]), but not significantly with the score for cartilage structure (P = 0.0557, r = 0.29). The interval between signals reflected well the actual thickness of the plug site. The significant relationships between ultrasound signal intensity and scores suggest that early degenerative changes in plug cartilage and cartilage-like tissue, especially in the superficial layer, are detectable by high-frequency ultrasound assessment

Single-photon generation from a nitrogen impurity center in GaAs

We have demonstrated single-photon emission from a nitrogen luminescence center in GaAs. An inhomogeneously broadened luminescence band formed by localized centers was observed in the spectral range from 1480 meV to 1510 meV at 5 K in nitrogen delta-doped GaAs. Optical properties of the individual centers were investigated by steady-state and time-resolved micro photoluminescence. We have found that a bright luminescence center emits single photons with a radiative lifetime of 650 ps, which is much shorter than the lifetime of NN pairs in previous reports