Some transformations and physicochemical properties of new S-substituted 5-thiophene-(3-ylmethyl)-4R-1,2,4-triazole-3-thiols

Nitrogen-containing heterocyclic compounds are widely represented in modern organic chemistry. A special place among the huge number of substances presented is occupied by derivatives of 1,2,4-triazole due to their uniqueness and prospects. In recent years, these derivatives have remained the object of close attention of both synthetic chemists and researchers engaged in the search for new biologically active substances of a wide spectrum of action, the synthesis of optical materials, dyes, corrosion inhibitors, veterinary drugs, etc. It is known from the literature that the combination of the aromatic heterocycle of thiophene and 1,2,4-triazole is a promising area for research. The result of such combinations is a number of compounds obtained with fairly high yields and a wide range of activities. Thus, further chemical modification of 1,2,4-triazole derivatives by additional administration of a typical thiophene pharmacophore is an urgent task. The aim of the study is to obtain new compounds in a number of S-substituted 5-thiophene-(3-ylmethyl)-4R-1,2,4-triazole-3-thiols and study their physico-chemical properties using complex analysis methods. The study of the physical and chemical properties of the compounds obtained was carried out according to methods that meet the standards of the state pharmacopoeia of Ukraine. The synthesis of new compounds was carried out according to generally accepted methods. The previously obtained 5-thiophene-(3-ylmethyl)-4h-1,2,4-triazole-3-thiol and 5-thiophene-(3-ylmethyl)-4-phenyl-1,2,4-triazole-3-thiol were used as starting compounds. Preparation of thioethanones was performed in an alkaline medium in the presence of methanol. The acids were obtained in an alcohol-alkaline medium with the addition of monochloroacetic acid. To obtain esters, two conventional methods were used. The first method is based on the alkylation in an alkaline-alcohol medium of the starting thiols with isopropyl ester of monochloroacetic acid. The second method is the classical esterification reaction. To confirm the structure and study of the physical and chemical properties, all the resulting compounds were crystallized. As a result, some new compounds that were not previously described were obtained, namely: thioethanones, thioacetamides, thioacetic acids, thioacetates, and a number of other thiopo-derivatives of these initial compounds. The structure and individuality of the obtained molecules were confirmed by elemental analysis (CHNS), IR and 1H NMR spectra. A number of new S-derivatives of 5-thiophene-(3-ylmethyl)-4R-1,2,4-triazole-3-thiols have been obtained. The physical and chemical properties of the obtained compounds are determined and their structure is confirmed using modern analysis methods. Some compounds will be transferred for further research.

Recombinant Production and One-Pot Purification for Enhancing Activity of Haloacid Dehalogenase from Bacillus cereus IndB1

In recent years we have witnessed the emergence of organohalogen utilization in various chemical-based industries, particularly polymer-based, agricultural, and pharmaceutical sectors. Despite this, organohalogen compounds are actually very dangerous to the environment, as they are difficult to be naturally degraded and generally toxic to organisms. A green and biocompatible method to overcome this issue is by employing enzymes that could convert organohalogens into non-toxic compounds, such as the class of enzymes known as haloacid dehalogenases. To enhance the activity of haloacid dehalogenase isolated from local strains of Bacillus cereus IndB1, we have developed a recombinant expression system using pET-bcfd1 plasmid in E. coli BL21 (DE3) host cells. Following enzyme production, we also demonstrated a one-pot purification system for the expressed dehalogenase, harnessing the presence of His-tag in the recombinant clones. Purification was carried out using Ni-NTA affinity column chromatography, using imidazole eluent with a concentration gradient of 10 mM to 500 mM. The enzyme activity was tested against the monochloroacetic acid (MCA) substrate according to the Bergmann and Sanik method, and the protein content in the solution was measured using the Bradford method. The purity of the enzyme after one-pot purification was confirmed by SDS-PAGE analyses, showing a single band of 40 kDa in size. Remarkably, the purified haloacid dehalogenase specific activity was increased by 12-fold compared to its crude enzyme extract. Therefore, the expression and purification system developed in this study allow further exploration of dehalogenases from local strains as an efficient catalyst for MCA biodegradation.Keywords: recombinant expression, haloacid dehalogenase, monochloroacetic acid, enzyme purification

Optimization of Monochloroacetic Acid Biodegradation by Recombinant E. coli BL21 (DE3)/pET-bcfd1 Carrying Haloacid Dehalogenase Gene from Bacillus cereus IndB1

In recent years, attention to microbial dehalogenase has continually increased due to its potential application, both in bioremediation and in the biosynthesis of fine chemicals. Many microbial recombinant strains carrying dehalogenase gene have been developed, particularly to increase the dehalogenase production and its quality. In this study, we aimed to find the optimum condition for the production of active haloacid dehalogenase by E. coli BL21 (DE3) harboring recombinant plasmid pET-bcfd1 that carried haloacid dehalogenase gene from Bacillus cereus IndB1 local strain. This would be examined by assessing the ability of whole cell life culture to degrade monochloroacetic acid (MCA) and quantifying the chloride ion released into the medium. Several variables were evaluated to find this optimal condition. We found that the best condition for MCA biodegradation using this recombinant clone was at 0.2 mM MCA, 10 μM of isopropyl β-D-1-thiogalactopyranoside (IPTG), 6 hours of pre-induction incubation at 37ºC with shaking, 2 hours IPTG induction at 30ºC with shaking, at pH 7 in Luria Bertani (LB) liquid medium without NaCl, which produced about 0.056 mM chloride ions. Inducer concentration, pre-induction incubation time and temperature, as well as induction time and temperature were apparent to be associated with the expression of the protein, while the MCA concentration and the pH of the medium influenced the ability of the recombinant E. coli BL21 (DE3)/pET-bcfd1 to grow in toxic environment. Our findings laid the foundation for exploration of dehalogenases from local Bacillus strains through genetic engineering for MCA biodegradation

Effect of Monochloroacetic Acid on Properties of Carboxymethyl Bacterial Cellulose Powder and Film from Nata de Coco

Nata de coco has been used as a raw material for food preparation. In this study, the production of carboxymethyl cellulose (CMC) film from nata de coco and the effect of monochloroacetic acid on carboxymethyl bacterial cellulose (CMCn) and its film were investigated. Bacterial cellulose from nata de coco was modified into CMC form via carboxymethylation using various concentrations of monochloroacetic acid (MCA) at 6, 12, 18, and 24 g per 15 g of cellulose. The results showed that different concentrations of MCA affected the degree of substitution (DS), chemical structure, viscosity, color, crystallinity, and morphology of CMCn. The optimum treatment for carboxymethylation was found using 24 g of MCA per 15 g of cellulose, which provided the highest DS at 0.83. The morphology of CMCn was related to DS value; a higher DS value showed denser and smoother surface than nata de coco cellulose. The various MCA concentrations increased the mechanical properties (tensile strength and percentage of elongation at break) and water vapor permeability of CMCn, which were related to the DS value.

Mixed esters of xanthogenic acids containing carbonyl, dithiocarbamine and thiocyane group and their research as additives improving tribological characteristics

On the basis of environmentally friendly raw materials of glycerol, by its interaction with monochloroacetic acid, 1,2,3-(trichloroacetyl)triglyceride was synthesized, which is used for further syntheses of xanthogenic acid derivatives containing a number of functional groups  dithiocarbamic, thiocyanic. The reaction of 1,2,3-(trichloroacetyl) triglyceride with potassium alkylxanthate in a ratio of 1:1 and 1:2 gave 1-butylxanthogenatoacetoxy-2,3-(dichloroacetoxy) propane and 1,3-di(alkylxanthogenatoacetoxy)-2-chloroacetoxypropane. Mixed esters of xanthogenic acid containing dithiocarbamic and thiocyanic groups were synthesized by the interaction of 1,2,3-(trichloroacetyl) triglyceride in the corresponding sequence, first with sodium diethyl dithiocarbamate, with potassium butylxanthate, as a result 1-butylxanthogenatoacetoxy-2-chloracetoxy-3-(diethyldithiocarbamatoacetoxy) propane. Sequential reaction of 1,2,3-(trichloroacetyl) triglyceride with potassium rhodanide with potassium butylxanthate gave 1-butylxanthogenatoacetoxy-2-chloroacetoxy-3-(thiocyanatoacetoxy)propane. The structure of the synthesized compounds has been proven by IR spectroscopy. The compounds have been tested as additives to improve the tribological characteristics of oils and have been shown to be effective.

Acid-base properties of fibrous polyampholytes with aminodiacetic functional groups

Polyampholytes, containing aminodiacetic functional groups, on the base of polyacrylonitrile fiber Nitron C were obtained by the amination reaction of nitrile groups with polyamines (number of repeating units from 1 to 5) and subsequent alkylation with monochloroacetic acid. The experimental results of potentiometric titration and sorption of supporting electrolyte (kCl) ions by synthesized ion exchangers were obtained, which made it possible to determine the acidity parameters of functional groups: the presence of four types of cation exchange and two types of anion exchange groups was detected in polyampholytes, the sequence of their neutralization in the titration process was established.

Skin burns from monochloroacetic acid leak in a chemical plant: a case report

The patient, a 45-year-old male chemical factory worker, was burned by monochloroacetic acid discharged from a ruptured pipe. The patient was merely flushed with water and did not leave the workplace immediately. As a result, he suffered local burn symptoms, which gradually worsened. Two and a half hours after the accident, he developed symptoms of systemic poisoning, such as lethargy and dyspnoea. After a thorough debridement of the wound surface and subsequent skin grafting combined with early glucocorticoid therapy and haemofiltration, a satisfactory result was achieved, and the patient eventually recovered. With the widespread use of monochloroacetic acid in China, incidents of poisoning with this chemical are becoming increasingly common, with more than 100 cases reported in the past ten years in China alone.