Use of wood vinegar to enhance 5-aminolevulinic acid production by selected Rhodopseudomonas palustris in rubber sheet wastewater for agricultural use

AbstractThis study aimed to produce inexpensive 5-aminolevulinic acid (ALA) in a non-sterile latex rubber sheet wastewater (RSW) by Rhodopseudomonas palustris TN114 and PP803 for the possibility to use in agricultural purposes by investigating the optimum conditions, and applying of wood vinegar (WV) as an economical source of levulinic acid to enhance ALA content. The Box–Behnken Design experiment was conducted under microaerobic-light conditions for 96h with TN114, PP803 and their mixed culture (1:1) by varying initial pH, inoculum size (% v/v) and initial chemical oxygen demand (COD, mg/L). Results showed that the optimal condition (pH, % inoculum size, COD) of each set to produce extracellular ALA was found at 7.50, 6.00, 2000 for TN114; 7.50, 7.00, 3000 for PP803; and 7.50, 6.00, 4000 for a mixed culture; and each set achieved COD reduction as high as 63%, 71% and 75%, respectively. Addition of the optimal concentration of WV at mid log phase at 0.63% for TN114, and 1.25% for PP803 and the mixed culture significantly increased the ALA content by 3.7–4.2times (128, 90 and 131μM, respectively) compared to their controls. ALA production cost could be reduced approximately 31times with WV on the basis of the amount of levulinic acid used. Effluent containing ALA for using in agriculture could be achieved by treating the RSW with the selected ALA producer R. palustris strains under the optimized condition with a little WV additive

Isolation of the cytotoxic constituent deoxypodophyllotoxin from the leaves of Juniperus chinensis

Deoxypodophyllotoxin was isolated as a cytotoxic constituent from ethanol extract of the leaves of Juniperus chinensis by assay-guided fractionation

Stabilization of cyclo (Gly-Tyr)-hydrolyzing Enzyme by Synthetic Glycoside-bearing Polymer

An enrichment technique led us to isolate 13 cyclo (Gly-Tyr)-assimilating bacteria, which had higher cyclo (Gly-Tyr)-hydrolyzing activity that the coryneform rod bacterium strain T-1-3-Y,previously isolated from soil. The cell-free extract of the strain Ol-L-2 prepared by ultrasonication in a buffer containing bovine serum albumin had cyclo (Gly-Tyr)-hydrolyzing activity while that of the strain T-1-3-Y showed no activity. GEMA (Glucosyloxyethyl methacrylate) polymer was found to be a more effective stabilizer for solubilizing cyclo (Gly-Tyr) hydrolase than bovine serum albumin.集積培養法を用いて新たに土壌よりcyclo(Gly-Tyr)資化性菌を分離し、その中から、従来の資化性菌より菌体当たりの活性の高い菌株を13株見いだした。これらの菌株を牛血清アルブミンを添加した緩衝液中に懸濁し、超音波破砕に供したところ、OI-L-2株由来の無細胞抽出液中にcyclo(Gly-Tyr)分解活性が検出された。この活性は安定化剤をアルブミンから配糖化合成高分子GEMAポリマーに代えることによりさらに安定に抽出された

An Analytical Method of lnophyllums A,B,C,D,E,and P,Anti - HIV Constituents of Calophyllum inophyllum by HPLC

In order to optimize cell culture conditions for producing inhibitors of HIV reverse transcriptase isolated from Calophyllum , inophyllum , a rapid and accurate anaalytical method for determining the quantity of the inhibitors, inophyllums A , B , C , D , E , and P was desired . We have established a quantitative analytical method using HPLC ,together with LCMS .最近、抗HIV活性化合物として注目されているCalophyllum inophyllumの葉の成分、inophyllum化合物を細胞培養で生産するためには、培養条件検討時に用いる当各成分の迅速かつ正確な分析法が必要である。本研究では、著者らの一人が以前に単離したinophyllum化合物のHPLC上の溶出位置を決定できることを示した

土壌から単離した真菌の培養物についての殺線虫活性のスクリーニングと殺線虫活性のある1菌株培養上清の予備分画

Cultures of 99 strains of fungi in potato-sucrose-malt extract medium were screened for nematicidal activity. Strong activity was found in the supernatant of the culture of fungus 18. Activity-guided preliminary fractionation of the supernatant showed that the active compound was neutral and water soluble.土壌から単離した99菌株の真菌の培養物について、以前の著者の1人が開発した線虫増殖阻害活性を検出するバイオアッセイを用いてスクリーニングを行った。1菌株の遠心分離上清に活性を認めた。予備的に行った各種クロマトグラフィーにおける挙動から、本活性化合物は、水溶性の中性化合物であると推定した

The use of rice straw broth as an appropriate medium to isolate purple nonsulfur bacteria from paddy fields

Abstract The aims were to explore an appropriate isolating medium for obtaining purple nonsulfur bacteria (PNSB) for use as biofertilizers in saline paddy fields and to obtain pure cultures. We therefore chose a defined isolating medium containing 0.25% NaCl, (Glutamate-Acetate broth, GA) and a rice straw broth to compare them for numbers of PNSB obtained, time to obtain pure cultures, diversity and costs. A total of 30 water and 30 sediment samples were collected from saline paddy fields in southern Thailand and used to isolate PNSB in both the isolating media. Based on 60 samples and a period of 13 days incubation under anaerobic light conditions, a greater number of samples produced PNSB growth in GA broth after only day 3; however, after that the rice straw broth provided about a 2 fold increase in the number of samples that produced PNSB growth. Colonies isolated from GA broth required a significantly higher number of repeated streaking to obtain a pure culture (average 3.5) than those from rice straw broth (average 2.7) and the latter medium also produced significantly (P < 0.05) more isolates per sample. Sixty samples of water and sediment, from rice paddies with salinity (average, 3.43 \ub1 0.67 mS/cm) and slight acidity (average, pH 5.84 \ub1 0.42) provided 62 PNSB isolates by GA broth and 210 isolates by rice straw broth, and rice straw broth also produced a greater prevalence of PNSB. Estimates of the costs based on current prices of media, Gas Pak and electricity to obtain PNSB with the use of GA broth was roughly 6 times higher than for the rice straw broth

マツノザイセンチュウ，Bursaphelenchus xylophilusに随伴する松萎凋性細菌の単離とその毒性代謝物質

Based on the observation that ray parenchyma cell death took place prior to nematode population increase in pine wood,the authors suspected that any microorganisms carried by pathogenic nematodes would be involvedin the pathogenic process.The authors screened pathogenic nematode-accompanying microbes for their toxicity against cultured cells of Pinus thunbergiis and isolated and identified 3 toxic strains,Bacillus cereus HY-3,B.subtilis HY-16,and Bmegaterium HY-17 , whose toxic products were identified as phenylacertic acid.Inoculation of pine seedlings with the toxic bacterium alone did not cause the seedling to wilt , but inoculation of pine seedlings with the toxic bacterium carried by weakly pathogenic nematodes wilted the seedling as much as strongly pathogenic nematodes . These results suggested that phenyllacetic acid-producing bacteria could invade pine trees by accompanying pine wood nematodes , and produce phenylacetic acid , a toxic metabolite , in the wood.マツ材線虫病の初期症状である樹脂道エピセリウム細胞や周辺柔細胞の変性は、マツノザイセンチュウのその場への到着および増殖に先行して起こることから、マツノザイセンチュウに随伴する微生物が発病に関与しているのだはないかと推論し、本研究では、クロマツ培養細胞に対する毒性を指標にして強病原生マツノザイセンチュウから毒物質生産細菌を数株を単離したえ。さらに、この毒性代謝物質を単離し、フェニル酢酸と同定した。フェニル酢酸生産細菌は、普遍的に存在するが、フェニル酢酸生産性には、大きい差があることを確認した。このフェニル酢酸生産細菌のみを接種しても、アカマツ実生は萎凋しなかったが、弱病原性マツノザイセンチュウに保持されて接種すると、ザイセンチュウの病原性が強くなった。このかとから、フェニル酢酸生産細菌は、マツノザイセンチュウに運搬されて松樹体内を移動し病原毒素フェニル酢酸を生産するものと推論した

Novel Cyclic Dipeptide Dehydrogenase and Assay Method for Activity

The cell-free extract prepared from cells of an albonoursin-producing actinomycete Streptomyces sp. KO2388 was found to catalyze the dehydrogenation of cyclo (L-Phe-L-Leu) (CFL) to albonoursin. This is the first report for the dehydrogenation at the α,β-positions of amino acid residues. The simple method for determining the dehydrogenation activity was devised by measuring the increase in UV absorption of the reaction mixture at 317nm, λmax(ε25,400) of albonoursin, where CFL had no absorption. Phenazine methosulfate was the most active cofactor for the dehydrogenation among several hydrogen acceptors.アルボノルシン生産菌、Streptomyces sp. KO2388株の無細胞抽出液が、cyclo(L-Phe-L-Leu)からアルボノルシンへの脱水素反応を触媒することを明らかにした。アミノ酸残基のα,β-位での脱水素反応を酵素レベルで明らかにしたのはこの報告が初めてである。本反応の簡便な測定法として、基質cyclo(L-Phe-L-Leu)には認められない。アルボノルシンの特異的な317nmにおける紫外吸収の増加を測定する方法を考案した。本方法を用いて数種の水素受容体を試験したところ、フェナジンメトサルフェートが最も有効であると判った

A Compound Which Can Be Used for Selecting Transfomed Plant Cells

Streptomyces sp,KBFP-2025株が生産するOxazolomycinは,Agrobacterium tumefaciensに対する特異的な抗菌活性により,ジャガイモのクラウンゴール形成を阻害する.さらに,本化合物はアルファルファの発芽を強く阻害し,ジャガイモ塊茎細胞を壊死させたが,ジャガイモのタラウンゴール細胞に対する生育阻害を示さなかった.すなわち,本化合物は非形質転換植物細胞に対する選択毒性を有しており,形質転換細胞の選抜に有用である

KCNJ13 Gene Deletion Impairs Cell Alignment and Phagocytosis in Retinal Pigment Epithelium Derived from Human-Induced Pluripotent Stem Cells

Purpose: The purpose of this study was to establish and analyze a cell model of Leber congenital amaurosis type 16 (LCA16), which is caused by mutations in the KCNJ13 gene encoding Kir7.1, an inward-rectifying potassium ion channel. Methods: The two guide RNAs specific to the target sites in the KCNJ13 gene were designed and KCNJ13 knock-out (KO) human-induced pluripotent stem cells (hiPSCs) were generated using the CRISPR/Cas9 system. The KCNJ13-KO hiPSCs were differentiated into retinal pigment epithelial cells (hiPSC-RPEs). The KCNJ13-KO in hiPSC-RPEs was confirmed by immunostaining. Phagocytic activity of hiPSC-RPEs was assessed using the uptake of fluorescently labeled porcine photoreceptor outer segments (POSs). Phagocytosis-related genes in RPE cells were assessed by quantitative polymerase chain reaction. Results: Most of the translated region of the KCNJ13 gene was deleted in the KCNJ13-KO hiPSCs by the CRISPR/Cas9 system, and this confirmed that the Kir7.1 protein was not present in RPE cells induced from the hiPSCs. Expression of RPE marker genes such as BEST1 and CRALBP was retained in the wild-type (WT) and in the KCNJ13-KO hiPSC-RPE cells. However, phagocytic activity and expression of phagocytosis-related genes in the KCNJ13-null hiPSC-RPE cells were significantly reduced compared to those of WT. Conclusions: We succeeded in generating an RPE model of LCA16 using hiPSCs. We suggest that Kir7.1 is required for phagocytosis of POSs by RPE cells and that impaired phagocytosis in the absence of Kir7.1 would be involved in the retinal degeneration found in LCA16