scholarly journals Optimization of Reaction Conditions for γ-Glutamylcysteine Production from Glutathione Using a Phytochelatin Synthase-Like Enzyme from Nostoc sp. Pasteur Culture Collection 7120

2021 ◽  
Vol 44 (12) ◽  
pp. 1832-1836
Author(s):  
Misa Muraoka ◽  
Moeka Ohno ◽  
Makoto Tateishi ◽  
Hideyuki Matsuura ◽  
Kazuya Nagano ◽  
...  
2005 ◽  
Vol 45 (3) ◽  
pp. 141 ◽  
Author(s):  
A-M. Vachot-Griffin ◽  
J. E. Thies

Monitoring the success of rhizobial inoculation requires reliable identification of the introduced strains in nodules and when recovered from field soil. The polymerase chain reaction (PCR) coupled with the use of either random or directed primers has increasingly become the molecular method of choice for characterising bacteria at the strain level. We have investigated the use of 5 markers (REP, ERIC, BOXA1R, RPO1 and IGS) commonly used for PCR fingerprinting to characterise rhizobia bacteria used in the manufacture of rhizobial inoculants in Australia. PCR with random primers often yields inconsistent results because most protocols do not specify stringent cycling and non-cycling parameters. We have increased the stringency and improved the specificity of reaction conditions for 4 of the 5 markers tested. Optimised protocols were then used to fingerprint the 39 strains of rhizobia bacteria held in the 1998 mother culture collection of the Australian Legume Inoculant Research Unit (ALIRU). Results for 34 strains using at least one marker are presented. Although the mother cultures of these inoculant strains undergo numerous quality assurance tests annually, it was not until PCR fingerprinting was applied that 2 strains, believed to be unique, were found to be identical. In the subsequent investigation, we determined that the 2 strains were originally unique but that a mix-up in the cultures had occurred at least 3 years before our analysis. Use of serology, plant infection tests and field tests were not sufficient to detect this problem. The use of PCR fingerprinting with optimised protocols has now been incorporated into the annual quality assurance regime used by the ALIRU who monitor strain quality for the Australian rhizobial inoculant industry. Higher quality rhizobial inoculant for use by Australian farmers is a beneficial outcome of this work.


2002 ◽  
Vol 364 (1) ◽  
pp. 129-136 ◽  
Author(s):  
Mahdi HEJAZI ◽  
Kirill PIOTUKH ◽  
Jens MATTOW ◽  
Rainer DEUTZMANN ◽  
Rudolf VOLKMER-ENGERT ◽  
...  

Recombinant plant-type asparaginases from the cyanobacteria Synechocystis sp. PCC (Pasteur culture collection) 6803 and Anabaena sp. PCC 7120, from Escherichia coli and from the plant Arabidopsis thaliana were expressed in E. coli with either an N-terminal or a C-terminal His tag, and purified. Although each of the four enzymes is encoded by a single gene, their mature forms consist of two protein subunits that are generated by autoproteolytic cleavage of the primary translation products at the Gly—Thr bond within the sequence GTI/VG. The enzymes not only deamidated asparagine but also hydrolysed a range of isoaspartyl dipeptides. As various isoaspartyl peptides are known to arise from proteolytic degradation of post-translationally altered proteins containing isoaspartyl residues, and from depolymerization of the cyanobacterial reserve polymer multi-l-arginyl-poly-l-aspartic acid (cyanophycin), plant-type asparaginases may not only function in asparagine catabolism but also in the final steps of protein and cyanophycin degradation. The properties of these enzymes are compared with those of the sequence-related glycosylasparaginases.


2021 ◽  
Vol 118 (40) ◽  
pp. e2106882118
Author(s):  
Jon S. Sauer ◽  
Ryan Simkovsky ◽  
Alexia N. Moore ◽  
Luis Camarda ◽  
Summer L. Sherman ◽  
...  

Algae cultivation in open raceway ponds is considered the most economical method for photosynthetically producing biomass for biofuels, chemical feedstocks, and other high-value products. One of the primary challenges for open ponds is diminished biomass yields due to attack by grazers, competitors, and infectious organisms. Higher-frequency observations are needed for detection of grazer infections, which can rapidly reduce biomass levels. In this study, real-time measurements were performed using chemical ionization mass spectrometry (CIMS) to monitor the impact of grazer infections on cyanobacterial cultures. Numerous volatile gases were produced during healthy growth periods from freshwater Synechococcus elongatus Pasteur Culture Collection (PCC) 7942, with 6-methyl-5-hepten-2-one serving as a unique metabolic indicator of exponential growth. Following the introduction of a Tetrahymena ciliate grazer, the concentrations of multiple volatile species were observed to change after a latent period as short as 18 h. Nitrogenous gases, including ammonia and pyrroline, were found to be reliable indicators of grazing. Detection of grazing by CIMS showed indicators of infections much sooner than traditional methods, microscopy, and continuous fluorescence, which did not detect changes until 37 to 76 h after CIMS detection. CIMS analysis of gases produced by PCC 7942 further shows a complex temporal array of biomass-dependent volatile gas production, which demonstrates the potential for using volatile gas analysis as a diagnostic for grazer infections. Overall, these results show promise for the use of continuous volatile metabolite monitoring for the detection of grazing in algal monocultures, potentially reducing current grazing-induced biomass losses, which could save hundreds of millions of dollars.


2007 ◽  
Vol 73 (23) ◽  
pp. 7605-7614 ◽  
Author(s):  
Sabrina Cadel-Six ◽  
Caroline Peyraud-Thomas ◽  
Luc Brient ◽  
Nicole Tandeau de Marsac ◽  
Rosmarie Rippka ◽  
...  

ABSTRACT Repeated dog deaths occurred in 2002, 2003, and 2005 after the animals drank water from the shoreline of the Tarn River in southern France. Signs of intoxication indicated acute poisoning due to a neurotoxin. Floating scum and biofilms covering pebbles were collected in the summers of 2005 and 2006 from six different sites along 30 km from the border of this river. The cyanobacterial neurotoxic alkaloid anatoxin-a and/or its methyl homolog, homoanatoxin-a, was detected in the extracts of most samples examined by gas chromatography-mass spectrometry. Fifteen filamentous cyanobacteria of the order Oscillatoriales were isolated and displayed four distinct phenotypes based on morphological characteristics and pigmentation. Three of the phenotypes can be assigned to the genus Oscillatoria or Phormidium, depending on the taxonomic treatises (bacteriological/botanical) employed for identification. The fourth phenotype is typical of the genus Geitlerinema Anagnostidis 1989. Eight strains rendered axenic were analyzed for production of anatoxin-a and homoanatoxin-a, and all strains of Oscillatoria/Phormidium proved to be neurotoxic. The genetic relatedness of the new isolates was evaluated by comparison of the intergenic transcribed spacer sequences with those of six oscillatorian strains from the Pasteur Culture Collection of Cyanobacteria. These analyses showed that the neurotoxic representatives are composed of five different genotypes, three of which correspond to phenotypes isolated in this study. Our findings prove that neurotoxic oscillatorian cyanobacteria exist in the Tarn River and thus were most likely implicated in the reported dog poisonings. Furthermore, they reemphasize the importance of monitoring benthic cyanobacteria in aquatic environments to fully assess the health risks associated with these organisms.


Author(s):  
James F. Hainfeld ◽  
Kyra M. Alford ◽  
Mathias Sprinzl ◽  
Valsan Mandiyan ◽  
Santa J. Tumminia ◽  
...  

The undecagold (Au11) cluster was used to covalently label tRNA molecules at two specific ribonucleotides, one at position 75, and one at position 32 near the anticodon loop. Two different Au11 derivatives were used, one with a monomaleimide and one with a monoiodacetamide to effect efficient reactions.The first tRNA labeled was yeast tRNAphe which had a 2-thiocytidine (s2C) enzymatically introduced at position 75. This was found to react with the iodoacetamide-Aun derivative (Fig. 1) but not the maleimide-Aun (Fig. 2). Reaction conditions were 37° for 16 hours. Addition of dimethylformamide (DMF) up to 70% made no improvement in the labeling yield. A high resolution scanning transmission electron micrograph (STEM) taken using the darkfield elastically scattered electrons is shown in Fig. 3.


2010 ◽  
Vol 1247 ◽  
Author(s):  
Rocío Calderón-Villajos ◽  
Carlos Zaldo ◽  
Concepción Cascales

AbstractControlled reaction conditions in simple, template-free hydrothermal processes yield Tm-Lu2O3 and Tm-GdVO4 nanocrystals with well-defined specific morphologies and sizes. In both oxide families, nanocrystals prepared at pH 7 reaction media exhibit photoluminescence in ∼1.95 μm similar to bulk single crystals. For the lowest Tm3+ concentration (0.2 % mol) in GdVO4 measured 3H4 and 3F4 fluorescence lifetimes τ are very near to τrad.


Author(s):  
Alex L. Bagdasarian ◽  
Stasik Popov ◽  
Benjamin Wigman ◽  
Wenjing Wei ◽  
woojin lee ◽  
...  

Herein we report the 3,5bistrifluoromethylphenyl urea-catalyzed functionalization of unactivated C–H bonds. In this system, the urea catalyst mediates the formation of high-energy vinyl carbocations that undergo facile C–H insertion and Friedel–Crafts reactions. We introduce a new paradigm for these privileged scaffolds where the combination of hydrogen bonding motifs and strong bases affords highly active Lewis acid catalysts capable of ionizing strong C–O bonds. Despite the highly Lewis acidic nature of these catalysts that enables triflate abstraction from sp<sup>2</sup> carbons, these newly found reaction conditions allow for the formation of heterocycles and tolerate highly Lewis basic heteroaromatic substrates. This strategy showcases the potential utility of dicoordinated vinyl carbocations in organic synthesis.<br>


2020 ◽  
Author(s):  
Alex L. Bagdasarian ◽  
Stasik Popov ◽  
Benjamin Wigman ◽  
Wenjing Wei ◽  
woojin lee ◽  
...  

Herein we report the 3,5bistrifluoromethylphenyl urea-catalyzed functionalization of unactivated C–H bonds. In this system, the urea catalyst mediates the formation of high-energy vinyl carbocations that undergo facile C–H insertion and Friedel–Crafts reactions. We introduce a new paradigm for these privileged scaffolds where the combination of hydrogen bonding motifs and strong bases affords highly active Lewis acid catalysts capable of ionizing strong C–O bonds. Despite the highly Lewis acidic nature of these catalysts that enables triflate abstraction from sp<sup>2</sup> carbons, these newly found reaction conditions allow for the formation of heterocycles and tolerate highly Lewis basic heteroaromatic substrates. This strategy showcases the potential utility of dicoordinated vinyl carbocations in organic synthesis.<br>


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