Adsorption ofCatharanthus roseus cells on various support particles

Summary Adsorption ofC. roseus cells producing indole alkaloids on various support particles were investigated in an attempt to find a suitable support material for surface culture of plant cells. Five different support particles namely gelatin, agar, alginate, polypropylene and glass beads were tested. Gelatin was found to be the most effective adsorbent resulting in nearly 30% adsorption of cells initially present in suspension. Adsorption isotherm of cells on gelatin beads was represented by a three parameter expression due to sigmoidal shape of the isotherm. The constants of the adsorption isotherm were determined using the experimental data.     

Identification of Mycoplasma Binding Proteins Utilizing A 100 Kilodalton Lung Fibroblast Receptor

Abstract

A 100 kilodalton glycoprotein receptor for Mycoplasma pneumoniae has been isolated from MRC-5 human lung fibroblasts. This receptor, as well as antireceptor serum, were both capable of inhibiting the attachment of ¹⁴C-labelled M. pneumoniae to MRC-5 fibroblasts. The receptor was also capable of inhibiting the attachment of C-labelled M. gallisepticum and M. genitalium, but not M. pulmonis, to MRC-5 fibroblasts. This indicates that a common sequence may exist in these binding proteins of M. pneumoniae, M. genitalium, and M. gallisepticum, This receptor and anti-receptor serum were utilized to probe M. pneumoniae, M. genitalium, and M. gallisepticum for their corresponding binding proteins. A 32 kilodalton protein in M. pneumoniae, a 90 kilodalton protein in M. genitalium and a 139 kilodalton protein in M. gallisepticum were recognized.     

Effects of agitation on enzymatic saccharification of cellulose

Summary Crystalline cellulose Avicel has been hydrolyzed byTrichoderma viride cellulase (Meicelase CEPB) under vaned agitation conditions and the effect of agitation on the adsorption of cellulase on cellulose has been studied. Agitation was found to enhance the hydrolysis pf crystalline cellulose; possibly the agitation enhances the adsorption of exoglucanase to shift the adsorption balance of exoglucanase and endoglucanase to a direction favorable for their synergistic action on the surface of cellulose.     

Effect of water on hydrolysis of olive oil by immobilized lipase in reverse phase system

Summary Candida rugosa lipase immobilized by adsorption on the swollen Sephadex LH-20 and Sephadex LH-60 could effectively hydrolyze olive oil at high concentration in a reverse phase system. Initial water content was found to be the most important factor that determines both the hydrolysis rate and the degree of hydrolysis; approximately 54% of water in the gels was readily utilized.     

Could the improvements in the alcoholic fermentation of high glucose concentrations by yeast immobilization be explained by media supplementation?

Summary The maximal concentration of ethanol produced during the fermentation of 320 g/l glucose bySaccharomyces bayanus was higher when the yeast cells were immobilized either by adsorption on celite or by entrapment in k-carrageenan beads (from 10.5% with free cells up to 14.5% and 13.1% (v/v) respectively). This increase was due to medium supplementation with the compounds present in the immobilization supports.     

Continuous high rate production of ethanol by Zymomonas mobilis in an attached film expanded bed fermentor

Summary Experiments were conducted with Zymomonas mobilis in an attached film expanded bed (AFEB) fermentor at different dilution rates, using a feed glucose concentration of 100 gm/l. Vermiculite was used as the bed material for attachment of the bacterial film. Ethanol volumetric productivities were maximum at a dilution rate of 3.6hr^-1. The productivities were 105 gm/l-hr and 210 gm/l-hr based on total fermentor volume and bed volume, respectively.     

Production of mold lactase

A process for production of mold lactase was developed. Tests were carried out in pilot and industrial scale with an Aspergillus niger strain selected after screening a number of molds.A computer coupled autoanalyzer system was used for monitoring enzyme formation in the pilot fermentor. Lactase production was investigated using different pH- and temperature-profiles. A. niger lactase has an acid pH optimum, a high temperature optimum and good stability. It does not require any metal ions. It is suitable for immobilization for hydrolysis of lactose in acid whey.Three-fold enhancement of lactase production was obtained by mutagenizing A. niger using NTG as mutagenic agent.The lactases produced by the mutants have the same pH and temperature optima and stability but the growth properties of the mutants were different from those of the original strain.Sufficient specific activity of the enzyme preparation for immobilization was obtained by purifying the enzyme by selective adsorption on Na-Ca-silicate.     

A rapid method for the purification of D-amino acid oxidase of Trigonopsis variabilis by hydrophobic chromatography

Summary A relatively simple method has been described for the rapid purification of D-amino acid oxidase from Trigonopsis variabilis by hydrophobic chromatography on Phenyl-Sepharose CL-4B and negative adsorption on DEAE-cellulose. The purified enzyme had a specific activity of 22–24 units at 25°C and exhibited three bands on enzymatic staining.     

In situ adsorption for enhanced alkaloid production byCatharanthus roseus

Summary A significant fraction (10–40%) of the indole alkaloids produced byCatharanthus roseus was observed to be secreted into the medium. When a neutral polymeric resin, known to adsorb these alkaloids, was added to the cultivation medium, the accumulation of total indole alkaloids and the specific alkaloids, ajmalicine and serpentine were stimulated. Sorbent addition was also observed to result in increased ratios of ajmalicine to serpentine, which suggests the potential of using in situ adsorption to direct metabolism toward a specific product or intermediate within a given pathway.     

VKCDB: Voltage-gated potassium channel database

Background  

The family of voltage-gated potassium channels comprises a functionally diverse group of membrane proteins. They help maintain and regulate the potassium ion-based component of the membrane potential and are thus central to many critical physiological processes. VKCDB (Voltage-gated potassium [K] Channel DataBase) is a database of structural and functional data on these channels. It is designed as a resource for research on the molecular basis of voltage-gated potassium channel function.     

Engineering Corynebacterium glutamicum with a comprehensive genomic library and phage-based vectors

The Gram-positive bacterium Corynebacterium glutamicum sustains the industrial production of chiral molecules such as L-amino acids. Through heterologous gene expression, C. glutamicum is becoming a sustainable source of small organic molecules and added-value chemicals. The current methods to implement heterologous genes in C. glutamicum rely on replicative vectors requiring lasting selection or chromosomal integration using homologous recombination. Here, we present a set of dedicated and transversal tools for genome editing and gene delivery into C. glutamicum. We generated a cosmid-based library suitable for efficient double allelic exchange, covering more than 94% of the chromosome with an average 5.1x coverage. We employed the library and an iterative marker excision system to generate the carotenoid-free C. glutamicum BT1-C31-Albino (BCA) host, featuring the attachment sites for actinophages ϕC31 and ϕBT1 for one-step chromosomal integration. As a proof-of-principle, we employed a ϕC31-based integration and a Cre system for the markerless expression of the type III polyketide synthase RppA, and a ϕBT1-based integration system for the expression of the phosphopantetheinylation-dependent non-ribosomal peptide synthetase BpsA in the C. glutamicum BCA host. The developed genomic library and microbial host, and the characterized molecular tools will contribute to the study of the physiology and the rise of C. glutamicum as a leading host for drug discovery.     

Biological oxidation of thianthrene,thioxanthene and dibenzothiophene by the thermophilic organismSulfolobus acidocaldarius

Summary Microbial oxidation of some model aromatic organic sulfur compounds such as thianthrene, thioxanthene and dibenzothiophene by the thermophilic organismSulfolobus acidocaldarius has been studied. Sulfate ions released as an oxidation product were measured to quantify the oxidations. The oxidation of the aforementioned refractory aromatic sulfur compounds byS. acidocaldarius may have applications in organic sulfur removal from hydrocarbon fuels such as coal and oil.     

Rapid screening of peptides for heavy metal binding

Summary A simple method that allows testing and characterisation of cadmium binding motifs by large set of immobilized peptides was developed. Hepta- and octapeptides containing cysteine and histidine residues were synthesized on cotton carrier. They were tested for cadmium binding. pH values of half metal dissociation were measured. Test for Cd binding was based on its precipitation with ferrous dipyridyl iodide. Subsequent dissolving of precipitate provided semiquantitative data on relative amount of Cd bound. Observed data for particular peptides corresponded with prediction.Abbreviations Boc tercial butyloxycarboyl - Fmoc fluorenylmethyloxycarboyl - tBu tercial butyl - DMF N,N-dimethylformamide - DCM dichlormethane - Me methyl     

PheMaDB: A solution for storage,retrieval, and analysis of high throughput phenotype data

Background  

OmniLog™ phenotype microarrays (PMs) have the capability to measure and compare the growth responses of biological samples upon exposure to hundreds of growth conditions such as different metabolites and antibiotics over a time course of hours to days. In order to manage the large amount of data produced from the OmniLog™ instrument, PheMaDB (Phenotype Microarray DataBase), a web-based relational database, was designed. PheMaDB enables efficient storage, retrieval and rapid analysis of the OmniLog™ PM data.     

Leghemoglobin: Different roles for different components?

The ratio of leghemoglobin c content to leghemoglobin a content in soybean (Glycine max) root nodules drops in the early stages of plant growth and becomes constant during flowering and fruiting. These data bring into question the assumption that leghemoglobin components have a single biochemical role in nitrogen fixation.     

An airlift-recycle fermenter for microbial desulfurization of coal

Summary An airlift-external recycle fermenter has been constructed and used for the removal of pyritic sulfur from coal samples (4% initial total sulfur) by the thermophilic, sulfur oxidizing organism Sulfolobus acidocaldarius. The airlift fermenter behaved as a well mixed reactor. Approximately, 30% of initial pyritic sulfur has been removed from a 5% coal slurry of ~125 particle size, at a maximum rate of 1.8 mg S/l.h.     

Rationale and design of a prospective study: Cervical Dystonia Patient Registry for Observation of OnaBotulinumtoxinA Efficacy (CD PROBE)

Background  

A registry of patients with cervical dystonia (Cervical Dystonia Patient Registry for Observation of onaBotulinumtoxinA Efficacy [CD PROBE]) was initiated to capture data regarding physician practices and patient outcomes with onabotulinumtoxinA (BOTOX^?, Allergan, Inc., Irvine, CA, USA). Methods and baseline demographics from an interim analysis are provided.     

MET-IDEA version 2.06; improved efficiency and additional functions for mass spectrometry-based metabolomics data processing

Metabolomics Ion-based Data Extraction Algorithm (MET-IDEA) is a computer program for processing large-scale metabolomics data. MET-IDEA utilizes network Common Data Form (netCDF) data files available from a diversity of chromatographically coupled mass spectrometry (MS) systems, utilizes the sensitivity and selectivity associated with selected ion quantification, and greatly reduces the time and effort necessary to obtain large-scale organized data. This article reports on recent improvements to MET-IDEA which include new visualization of peak integrations, display of mass spectra associated with integrated peaks, and optional manual peak integration. The computational performance of MET-IDEA has also been improved to avoid memory overflow during the processing of large data sets and the software made compatible with 64 bit CPUs and operating systems. These new functions improve the performance of MET-IDEA, and they allow users to visualize peak integrations and curate the results through manual integration if desired. The improved version of MET-IDEA better facilitates the quantitative analysis of complex MS-based metabolomics data. MET-IDEA is freely available for academic and non commercial use at (http://bioinfo.noble.org/gateway/index.php?option=com_wrapper&;Itemid=57). Commercial use is available via licensing agreement.

     

Synthesis and Biological Evaluation of Some Acyclic Pyridine C-Nucleosides. Part Two

Abstract

3-Bromo-5-(2-hydroxyethylthiomethyl)pyridine (7) was synthesized by reaction of 3-bromo-5-chloromethylpyridine hydrochloride (6) with the mono sodium salt of 2-mercaptoethanol. 3-Bromo-5-hydroxymethylpyridine (10) was, after protection as a silyl ether, converted to the 3-carboxy analogue using BuLi and CO₂. After deprotection with NH₄F, the alcohol function was chlorinated using SOCl₂. Finally, attachment of the acyclic chain and ammonolysis gave the acyclic nicotinamide nucleosides. Treatment of the latter compounds with Lawesson's reagent gave the thioamide analogues. All compounds were identified by NMR and DCI-MS. The acyclic pyridine C-nucleosides were evaluated against a series of tumor-cell lines and a variety of viruses. No marked biological activity was found.     

Stereochemistry of naturally-occurring 25-hydroxyvitamin D3-26,23 lactone as determined by radioligand binding analysis and high-performance liquid chromatography

The four stereoisomers of 25-hydroxyvitamin D₃-26,23 lactone (25-OHD₃-26,23 lactone) were tested against in vivo 25-OHD₃-26,23 lactone to determine their relative competition in the radioligand binding assays for 25-OHD₃ and 1,25-(OH)₂D₃. The 25R-OHD₃-26,23S lactone and in vivo 25-OHD₃-26,23 lactone behaved identically in the radioligand binding assay for 25-OHD₃ and were ～5-fold more potent than 25-OHD₃ at displacing 25-OH[³H]D₃. The 25S-OHD₃-26,23S lactone was the poorest competitor in this assay, requiring a 10-fold excess relative to 25-OHD₃ to displace 50% of the 25-OH[³H]D₃. The order of competition in the 25-OHD₃ radioligand binding assay was 25R-OHD₃-26,23S lactone = in vivo 25-OHD₃-26,23 lactone ? 25S-OHD₃-26,23R lactone > 25-OHD₃ ? 25R-OHD₃-26,23R lactone > 25S-OHD₃-26,23S lactone. The order of competition in the 1,25-(OH)₂D₃ cytosol receptor assay was essentially reversed from the competition in the 25-OHD₃ assay and was 25S-OHD₃-26,23S lactone > 25-OHD₃ ? 25S-OHD₃-26,23R lactone > 25R-OHD₃-26,23S lactone = in vivo 25-OHD₃-26,23 lactone. When tested in a high-performance liquid chromatographic system which separates all four stereoisomers, the in vivo 25-OHD₃-26,23 lactone comigrated with synthetic 25R-OHD₃-26,23S lactone. These data firmly establish that the naturally-occurring 25-OHD₃-26,23 lactone has the 25R, 23S stereochemistry. In addition, these data are the first to demonstrate that the four stereoisomers of 25-OHD₃-26,23 lactone have different affinities for the plasma vitamin D binding protein and the 1,25-(OH)₂D cytosol receptor.