Diethyl Sulfate Research Articles

Revealing contaminants in China's recycled PET: Enabling safe food contact applications

The lack of data on chemical contamination in recycled polyethylene terephthalate (rPET) in food contact applications, impedes the legalization of rPET in food contact applications in China. This study offers a comprehensive analysis of contaminants in hot-washed rPET flakes, identifying 1011 compounds. While most substances were detected infrequently, some i.e., bis(2-ethylhexyl) phthalate and mineral oil aromatic hydrocarbons stood out for their relatively high average concentrations. Limonene emerged as the most frequently detected flavorants. Rare genotoxic compounds like safrole and diethyl sulfate were detected at low levels, emphasizing the importance of adopting a conservative evaluation approach. Molecular descriptor analysis guided the proposal of four new surrogates, i.e., dimethyl pentanedioate, 2-tridecanone, bis(2-ethylhexyl) terephthalate, and 2,2-dimethoxy-1,2-diphenylethanone, for challenge tests, aiming to better capture the chemical diversity of rPET contaminants. This research unveils the contamination profile of Chinese rPET and suggests adjustments to evaluation protocols, potentially facilitating its high-value applications and minimizing potential harms.

Effect of Treatments of Colchicine, Diethyl Sulphate and Triethylamine in Tomato (<I>Lycopersicon esculentum</I> Mill.)

The present investigation was undertaken in one variety of Tomato, Best of All, to compare the cytogenetic effects of treatments of seeds of tomato with colchicine, diethyl sulphate and triethylamine and also to compare the morphological and physiological effects of these chemicals.Lowest number of seeds germinated from diethyl sulphate treatment. Plants originating from colchicine, diethyl sulphate and triethylamine treatments showed many variant types. Interesting among these were those showing spotting on leaves originating from diethyl sulphate treatment, dwarf ones originating from colchicine and diethyl sulphate treatments and those with abnormal leaves originating from treatment with all the three chemicals.Mitotic irregularities included clumped chromosomes and fragments at metaphase and bridges and fragments at anaphase while in meiosis clumped chromosomes, univalents, trivalents and quadrivalents were noted at metaphase I. Anaphase I irregularities included cells with fragments, bridges with or without fragments, lagging chromosomes and unequal separation of chromosomes. In metaphase II cells with aneuploid chromosome numbers and those with fragments were noticed while in anaphase II cells were found with fragments and unequal separation of chromosomes.The importance of detecting interesting mutants in the Ma generation and the importance of finding translocations and inversions have been pointed out.

A Novel Preparation and Purification of Ethyl-β-Cyclodextrins

Ethyl-β-cyclodextrin can be widely used for controlled release of drugs, but its application is greatly limited due to the use of toxic ethylation reagents such as diethyl sulfate, chloroethane, and iodoethane in previous synthesis processes. This article expands the application range of ethyl-β-cyclodextrin by using green ethylation reagent instead of previously toxic ethyl reagents to synthesize ethyl-βcyclodextrin. Ethyl-β-cyclodextrins were studied by reaction with diethyl carbonate and β-cyclodextrin in DMF using anhydrous potassium carbonate as catalyst, During the reaction process, in order to avoid oxidation, the samples were kept in a protective atmosphere of N<sub>2</sub> flow. A new green synthesis process for ethyl-cyclodextrins, using silica gel chromatography to synthesize and purify four substituted 6-O-ethyl-cyclodextrins. Their structures were characterized by IR, MS, <sup>1</sup>H-NMR, and <sup>13</sup>C-NMR. The final product with a yield of 4% was obtained by chromatographic separation using acetonitrile-concd aq NH<sub>3</sub>-H<sub>2</sub>O-EtOAc (6:1:3:1) as the eluent. IR data indicate that the obtained product is the expected product. <sup>13</sup>C-NMR characterization indicates that the substitution position of the product is at position 6, <sup>1</sup>H-NMR, and MS characterization indicate that the degree of substitution of the product is 4. This synthesis method takes full advantage of the spatial selectivity of β-cyclodextrin. The method is green and simple. The target product is synthesized in one step, which is superior to previous reports.

In vitro induction and investigation of salinity tolerance in the callus of Vigna spp. (L)

Cowpeas can be employed for rotational grazing or generating high-quality food when paired with crops such as maize. Cowpea seeds provide a substantial amount of protein and other essential elements. The purpose of this work was to use a mutagen and a high salt concentration to extract a salt tolerance gene from the callus. The callus was induced by adding 2,4-Dichlorophenoxyacetic acid 2,4- D at mg/l and was then subjected to various concentrations of a combination of salts consisting of sodium chloride NaCl and Calcium Chloride.CaCl2, in addition to Magnesium chloride MgCl2, with a rate of 2:2:1 and potion 0, 50, 100, 150, 200, and 250 mM of the salt mixture added to the culture medium. Various levels of di ethyl sulfate (DES) mutagenic solution at a concentration of 0.1 mM were used to soak the callus for 30 minutes. The results showed that when using quantitative polymerase chain reaction q-PCR, they showed two genes of vigna, the first gene in this research. Aldehyde dehydrogenase family 3 (ADF3) demonstrated the explanation of the genes at doses of DES+200, 200, and 250 mM; however, the most elevated level of gene expression at DES+200mM is CT 30.47, with an upregulation of 2.479. The second gene, Protein S2, demonstrated that the gene is highly expressed at DES+200mM, cycle threshold (CT) 31.68, and DES+250mM concentrations and is upregulated at DES+150mM was 128.

Enhanced Production of Sisomicin in Micromonospora inyoensis by Protoplast Mutagenesis and Fermentation Optimization.

Sisomicin is a broad-spectrum aminoglycoside antibiotic and is the precursor of netilmicin and plazomicin. However, the fermentation level of sisomicin is still low compared with other antibiotics, which restricts the application of sisomicin and its derivatives. In this study, to improve sisomicin production, breeding of high-yielding sisomicin strains was conducted with chemical mutagenesis using Micromonospora inyoensis OG-1 (titer, 1042 U·mL-1) as the starting strain. Protoplast preparation was conducted under optimal conditions, and protoplast mutagenesis was performed with a suitable concentration of diethyl sulfate. Subsequently, a high-yielding and genetically stable strain (H6-32) was obtained by screening, with a sisomicin titer of 1486 U·mL-1 (an increase of 42.6%). Finally, carbon and nitrogen sources were optimized to further improve sisomicin production, and a sisomicin titer of 1780 U·mL-1 was ultimately obtained by controlling the dissolved oxygen level at 30% in a 5-L fermenter, which to the best of our knowledge is the highest reported titer ever achieved by fermentation. Comparative genome analysis showed that a total of 13 genes in the genome of the mutant strain H6-32 were mutated compared to the original strain. This study not only provides a reference for further breeding of high-yielding strains and fermentation optimization, but also enhances our understanding of sisomicin production.

Compound mutation by ultraviolet and diethyl sulfate of protease producing thermophilic bacteria to hydrolyze excess sludge

Excess sludge (ES), a resource-rich organic waste, can be solubilized by thermophilic enzymes to extract proteins for sludge reduction and resources recovery. To solve the problems of low hydrolysis effect of ES and low enzyme producing ability of wild thermophilic bacteria, ultraviolet and diethyl sulfate (UV-DES) were adopted to mutate thermophilic bacteria in this study. Mutation sites were detected and annotated by whole genome sequencing analysis. The results showed that UV-DES mutagenesis could effectively improve enzyme-producing capacity of thermophilic bacteria and promote the hydrolysis of ES. The protease activity of the mutant strain KT16 was 46.7 % higher than that of the original strain DC8. The protein extraction rate with enzyme produced by KT16 reached 83.3 %. The total content of proteins recycled through KT16 enzyme solution was 3539.6 mg·L-1, 18.4 % higher than that of DC8. This work provided a theoretical idea and technical guidance for the protein recovery from ES.

Phytochemical profiling, toxicity studies, wound healing, analgesic and anti-inflammatory activities of Musa paradisiaca L. Musaceae (Plantain) stem extract in rats

Ethnopharmacological relevanceThe stem of Musa paradisiaca (plantain) has found application in traditional medicine for the treatment of diabetes, inflammation, ulcers and wound injuries. Aim of the studyThis study investigated the phytochemical composition, toxicity profile, wound healing, anti-inflammatory and analgesic effects of aqueous Musa paradisiaca stem extract (AMPSE) in rats. MethodsPhytochemical analysis of methanol-MPSE was performed by gas chromatography-mass spectrometry (GC-MS). Acute toxicity testing was carried out through oral administration of a single dose of AMPSE up to 5 g/kg. Four separate groups of rats were used for the subacute toxicity testing (n = 6). Group 1 served as a normal control and did not receive AMPSE, groups 2–4 received AMPSE daily by gavage for 28 days. In the experiments with excision and incision wounds, the rats were treated with 10 w/w AMPS extract. The anti-inflammatory and analgesic effects of AMPSE were assessed using egg albumin-induced paw oedema and acetic acid-induced writhing methods, respectively. For the subacute, anti-inflammatory and analgesic studies, AMPSE was administered to the experimental rats at doses of 300, 600 and 900 mg/kg body weight. ResultsBioactive compounds identified include β-sitisterol, n-hexadecanoic acid, octadecanoic acid, diethyl sulfate, p-hydroxynorephedrine, phenylephrine, nor-pseudoephedrine, metaraminol, pseudoephedrine and vanillic acid. No signs of toxicity and no deaths were observed in all the groups. For the groups treated with AMPSE for 28 days, a significant reduction in alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, urea, sodium, chloride, total cholesterol, triglycerides, and low-density lipoprotein cholesterol were observed while high density lipoprotein cholesterol, glutathione and superoxide dismutase increased compared to control (p < 0.05). In wound healing experiments, AMPSE showed greater percent wound contraction and wound resistance fracture compared to the povidone-iodine (PI) treated and control groups. Treatment with 900 mg/kg AMPSE resulted in significant (p < 0.05) anti-inflammatory and analgesic effects compared to the control. ConclusionThis study shows that AMPSE is not toxic but contains biologically active compounds with hepatoprotective, anti-inflammatory, lipid-lowering and wound-healing effects. Treatment of rats with AMPSE has shown that AMPSE has anti-inflammatory, analgesic, hepatoprotective, lipid-lowering and wound-healing effects, supporting its therapeutic use in ethnomedicine.

Evaluation of mutagenic samples of winter garlic

Relevance. The use of chemical mutagenesis on garlic is relevant for the creation of new forms with stable characteristics, better biochemical qualities, increased yield, for use in breeding practice.Results. The reaction of garlic treated with two chemical mutagens was evaluated: DES (diethyl sulfate) – 0.025; 0.05 and 0.1% (mutants 2, 3 and 4), as well as DMS (dimethyl sulfate) – 0.02; 0.04 and 0.08% solutions (mutants 5, 6 and 7). The aftereffect of chemical mutagens was noted with low and medium variability of signs in the second–fifth generation. According to the low variability of 1.6–9.4%, the height of plants (samples M 3 and 6), the number of leaves (M 5), the height of the false stem (M 3), the diameter of bulbs (M 6), as well as the length of leaves and the weight of bulbs samples M 3, 5 and 6 were distinguished. The effect of mutagens on morphometric signs of garlic were also tracked in the second generation (re-growing from air bulbs), where the parameters of low and medium variability of signs are marked. The majority of samples on 7 signs reflected a low variability of 5.5–10.0% and only 2 mutants showed an average variability of 11.4–16.4%: M 6 in plant height and false stem and M 3 in bulb diameter and yield. The correlation between their value in the 5th generation and in the 2nd generation also revealed strong direct and average connections. High heritability of h2 signs = 0.74–0.99 for DES preparations – 0.05%, VMI – 0.02 and 0.04%, was the most important indicator of the selection of valuable signs.

Transcriptome Analysis Reveals an Eicosapentaenoic Acid Accumulation Mechanism in a Schizochytrium sp. Mutant.

Eicosapentaenoic acid (EPA) is an omega-3 long-chain polyunsaturated fatty acid (PUFA) essential for human health. Schizochytrium is a marine eukaryote that has been widely utilized for the synthesis of PUFAs. The current low potency and performance of EPA production by fermentation of Schizochytrium spp. limits its prospect in commercial production of EPA. Since the synthesis pathway of EPA in Schizochytrium spp. is still unclear, mutagenesis combined with efficient screening methods are still desirable. In this study, a novel screening strategy was developed based on a two-step progressive mutagenesis method based on atmospheric and room temperature plasma (ARTP) and diethyl sulfate (DES) after multiple stresses (sethoxydim, triclosan and 2,2'-bipyridine) compound screening. Finally, the mutant strain DBT-64 with increased lipid (1.57-fold, 31.71 g/L) and EPA (5.64-fold, 1.86 g/L) production was screened from wild-type (W) strains; the docosahexaenoic acid (DHA) content of mutant DBT-64 (M) was 11.41% lower than that of wild-type strains. Comparative transcriptomic analysis showed that the expression of genes related to the polyketide synthase, fatty acid prolongation, and triglyceride synthesis pathways was significantly upregulated in the mutant strain, while the expression of genes involved in the β-oxidation pathway and fatty acid degradation pathway was downregulated in favor of EPA biosynthesis in Schizochytrium. This study provides an effective strain improvement method to enhance EPA accumulation in Schizochytrium spp. IMPORTANCE Schizochytrium, a marine eukaryotic microorganism, has emerged as a candidate for the commercial production of PUFAs. EPA is an omega-3 PUFA with preventive and therapeutic effects against cardiovascular diseases, schizophrenia, and other disorders. Currently, the low potency and performance of EPA production by Schizochytrium spp. limits its commercialization. In this study, we performed two-step progressive mutagenesis based on ARTP and DES and screened multiple stresses (sethoxydim, triclosan, and 2,2'-bipyridine) to obtain the EPA-high-yielding Schizochytrium mutant. In addition, high expression of the polyketide synthase pathway, fatty acid elongation pathway, and triglyceride synthesis pathway in the mutants was confirmed by transcriptomic analysis. Therefore, the multistress screening platform established in this study is important for breeding EPA-producing Schizochytrium spp. and provides valuable information for regulating the proportion of EPA in microalgal lipids by means of genetic engineering.

Mutagenesis of high-efficiency heterotrophic nitrifying-aerobic denitrifying bacterium Rhodococcus sp. strain CPZ 24

Breeding high-efficiency heterotrophic nitrifying–aerobic denitrifying (SND) bacteria is important for the removal of biological nitrogen in wastewater treatment. In this study, a high-efficiency SND mutant strain, ΔRhodococcus sp. CPZ 24, was obtained by ultraviolet–diethyl sulfate compound mutagenesis. The maximum nitrification and denitrification rates were 3.77 and 1.37 mg·L−1·h−1, respectively 30.30 % and 17.10 % higher than those of wild bacteria. Biolog technology and network model analysis revealed that ΔCPZ 24 significantly improved the utilisation ability and metabolic activity of organic carbon sources. Furthermore, the expression levels of the nitrogen removal function genes nxrA, nosZ, amoA, and norB in strain ΔCPZ 24 increased significantly. In actual sewage, mutant bacteria ΔCPZ 24 have a 95.05 % ammonia–nitrogen degradation rate and a 96.67 % nitrate-nitrogen degradation rate. These results suggested that UV-DES compound mutation was a successful strategy to improve the nitrogen removal performance of SND bacteria in wastewater treatment.

Comparative study on the effect of chemical mutagens of sodium azideand di-ethyl sulfate on improving morphological traits and yield components of Borgo officinalis L, plant

Borage seeds were soaked in different doses of sodium azide (SA) and di-ethyl sulphate (DES), at 0.1,0.2,0.3 and 0.4% for 6 hrs., M2 seeds were harvested from M1 plants.In MG1 it was found that most SA doses increased plant height and stem thickness. SA at 0.3% &amp; 0.4%, increased the ramification of main branches and enhanced leaves formation, leaf area, FW and DW of leaves, DW /FW ratios and root length. SA 0.3% was more effective than 0.4% and SA 0.2% increased the root number. Whereas, in MG2, SA 0.3 % significantly increased most morphological traits, but stem thickness responded well to SA at 0.1% and No. of branches was increased with SA 0.4 %. A great improvement in the formation of leaves obtained with SA 0.3%.The leaves DW/FW ratios, showed a positive response to SA 0.1 and 0.3% Concerning DES effects in MG1 and 2, the results in MG1revealed that all doses increased plant height and stem diameter and 0.4% DES produced the tallest and highest No. of branches and at 0.1% gave thickest stems . Applying DES 0.3% greatly improved the formation of leaves and the largest leaf area was recorded with 2000 ppm DES.

Arthrospira platensis Mutagenesis for Protein and C-Phycocyanin Improvement and Proteomics Approaches.

Arthrospira (Spirulina) platensis is known for its use as a food supplement, with reported therapeutic properties including antiviral, anti-inflammatory and antioxidant activity. Arthrospira is also an excellent source of proteins and C-phycocyanin. The latter is a light-harvesting pigment-protein complex in cyanobacteria, located on the outer surface of the thylakoid membrane and comprising 40 to 60% of the total soluble protein in cells. Random mutagenesis is a useful tool as a non-genetically modified mutation method that has been widely used to generate mutants of different microorganisms. Exposure of microalgae or cyanobacteria to chemical stimuli affects their growth and many biological processes. Chemicals influence several proteins, including those involved in carbohydrate and energy metabolisms, photosynthesis and stress-related proteins (oxidative stress-reactive oxygen species (ROS) scavenging enzymes). Signal transduction pathways and ion transportation mechanisms are also impacted by chemical treatment, with changes causing the production of numerous biomolecules and stimulation of defence responses. This study compared the protein contents of A. platensis control and after mutagenesis using diethyl sulphate (DES) under various treatment concentrations for effective mutation of A. platensis. Results identified 1152 peptides using proteomics approaches. The proteins were classified into 23 functional categories. Random mutagenesis of A. platensis by DES was found to be highly effective for C-phycocyanin and protein production.

Morphologically favorable mutant of Trichoderma reesei for low viscosity cellulase production.

Metabolite production by filamentous fungi hampered because of high viscosity generated during growth. Low viscosity fermentation by mold is one of the preferred ways of large scale enzymes production. Cellulolytic enzymes play a key role during the process of lignocellulosic biomass conversion. In this study, a mutant RC-23-1 was isolated through mutagenesis (diethyl sulfate followed by UV) of Trichoderma reesei RUT-C30. RCRC-23-1 not only gave higher cellulase production but also generated lower viscosity during enzyme production. Viscosity of mutant growth was more than three times lower than parent strain. RC-23-1 shows unique, yeast-like colony morphology on solid media and small pellet-like growth in liquid media. This mutant did not spread like mold on solid media. This mutant produces cellulases constitutively when grown in sugars. Using only glucose, the cellulase production was 4.1 FPU/ml. Among polysaccharides (avicel, xylan, and pectin), avicel gave maximum of 6.2 FPU/ml and pretreated biomass (rice straw, wheat straw and sugarcane bagasse) produced 5.1-5.8 FPU/ml. At 7 L scale reactor, fed-batch process was designed for cellulase production using different carbon and nitrogen sources. Maximum yield of cellulases was 182 FPU/g of lactose consumed was observed in fed-batch process. The produced enzyme used for hydrolysis of acid pretreated rice straw (20% solid loading) and maximum of 60% glucan conversion was observed. RC-23-1 mutant is good candidate for large scale cellulase production and could be a model strain to study mold to yeast-like transformation.

EFFECT OF DIETHYL SULPHATE ON M1 ATTRIBUTES AND SDS-PAGE ANALYSIS IN GROUNDNUT (Arachis hypogaea L.)

The present study was carried out to know the effect of diethyl sulphate on groundnut cultivar TMV-7 particularly in M1 generation. The healthy and viable seeds of groundnut cultivar TMV-7 were exposed to different concentrations of diethyl sulphate (5 to 25 mM) and sown in the field. The various growth and morphological parameters like percentage of seed germination, survival percentage plant height, days to flowering, total height at time of harvest, root length and number of lateral roots, number of pods per plant and pod weight were decreased with increasing concentrations of diethyl sulphate. All the characteristics studied were inhibited when compared to control. The lower concentrations such as 5, 10 and 15 mM concentrations enhanced all the parameters. But there was a sudden decrease in 20 and 25 mM. The various concentrations of DES played a different role in the field condition itself. The shoots developed from different concentrations of DES showed variation in their number and height. The protein content also showed decreasing trend with increasing concentrations of DES. There was a steady increase of protein content up to 15 mM DES. After that, there was a drastic reduction in the protein content in 20 and 25 mM. From the diethyl sulphate treated seeds, the embryo axes were collected and subjected to SDS-PAGE analysis. The protein banding pattern also differentiated among various concentrations of DES. The protein band with 77.5 kDa was observed in 5 and 10mM treatments. Like that the 13.5 kDa was present in 10 and 25mM treatments. The 48.25 kDa recorded in 15 and 20 mM treatments.

Enhancing Ethanol Tolerance via the Mutational Breeding of Pichia terricola H5 to Improve the Flavor Profiles of Wine

Although using non-Saccharomyces yeasts during alcoholic fermentation can improve the wine aroma, most of them are not ethanol tolerant; therefore, in 2017, this study screened 85 non-Saccharomyces yeasts isolated and identified from 24 vineyards in seven Chinese wine-producing regions, obtaining Pichia terricola strain H5, which displayed 8% ethanol tolerance. Strain H5 was subjected to ultraviolet (UV) irradiation and diethyl sulfate (DES) mutagenesis treatment to obtain mutant strains with different fermentation characteristics from the parental H5. Compared with strain H5, the UV-irradiated strains, UV5 and UV8, showed significantly higher ethanol tolerance and fermentation capacity. Modified aroma profiles were also evident in the fermentation samples exposed to the mutants. Increased ethyl caprate, ethyl caprylate, and ethyl dodecanoate content were apparent in the UV5 samples, providing the wine with a distinctly floral, fruity, and spicy profile. Fermentation with strain UV8 produced a high ethyl acetate concentration, causing the wine to present a highly unpleasant odor. To a certain extent, UV irradiation improved the ethanol tolerance and fermentation ability of strain H5, changing the wine aroma profile. This study provides a theoretical basis for the industrial application of non-Saccharomyces yeasts that can improve wine flavor.

Enhanced antimicrobial activity against Alicyclobacillus acidoterrestris in apple juice by genome shuffling of Lactobacillus acidophilusNX2‐6

AbstractFood safety and quality are seriously compromised by the growth of spoilage bacteria in food. Various lactic acid bacteria (LAB) may inhibit the growth of spoilage bacteria in food and animal feed. In our study, the antibacterial activity of Lactobacillus acidophilus NX2‐6 was improved by genome shuffling. The starting populations were obtained via 60Coγ‐ray, diethyl sulfate mutagenesis, and UV mutagenesis of protoplasts. The optimal conditions for protoplast formation and regeneration were found to be 30 mg/ml of lysozyme at 37°C for 40 min. The protoplasts were inactivated by UV irradiation for 110 s or by heating at 58°C for 30 min. After two rounds of genome shuffling, the inhibitory activity of strain F50 was improved by 5.6‐fold compared to the original strain. The shuffled strain has broad‐spectrum antibacterial activity and is a promising candidate for bio‐preservative. The antibacterial substances produced by L. acidophilus NX2‐6 can be applied to the preservation of apple juice. The results showed that the growth of Alicyclobacillus acidoterrestris in apple juice was significantly inhibited by the addition of 0.1% culture supernatant with acidocin NX2‐6 at 28°C and 0.2% culture supernatant with acidocin NX2‐6 at 45°C for 30 days. The sugar−acid ratio, pH value, clarity, and fruit flavor of apple juice were well maintained during storage. This study showed that genome shuffling was an effective strategy to improve the bacterial inhibitory activity of LAB and that the shuffled LAB and their metabolites are expected to be used as bio‐based food protectors for food safety.

Screening of active sites and study on immobilization of Bacillus cereus phospholipase C

Bacillus cereus phospholipase C (PLCBC) was obtained by treating Bacillus cereus with ultraviolet light and diethyl sulfate. The active centers of PLCBC were determined, and its inactive center groups were combined with the carboxyl modified carrier to obtain immobilized PLCBC. The enzyme activity was 289 U/mg. The active group of the enzyme was –COOH, while the binding site was –NH2. The –COOH and –NH2 of PLCBC were chemically modified. The immobilization sites –COOH and –NH2 were used to covalently bind to the carrier, and the enzyme activity recovery ratio was 59.0% and 95.0%, respectively, with Vmax and Michaelis constant of the latter being closer to the free PLCBC. The carrier and immobilized enzyme were characterized. The optimal pHs for the free and immobilized PLCBC were 7.0 and 7.5, respectively, and the optimal temperatures were 50 °C and 55 °C, respectively. After 7 cycles, the relative activity of PLCBC–Fe3O4/SiO2/HPG–COOH was 87%. In this work, a new type of immobilized PLCBC was prepared through active site screening and oriented immobilization. It provided a theoretical basis for the application of immobilized phospholipase in industrial oil degumming.

Crystal structure and Hirshfeld surface analysis of 1-[(benzyl-dimethyl-sil-yl)meth-yl]-1-ethyl-piperidin-1-ium ethane-sulfonate.

The title mol-ecular salt, C17H30NSi+·C2H5O4S-, belongs to the class of a-amino-silanes and was synthesized by the alkyl-ation of 1-[(benzyl-dimethyl-sil-yl)meth-yl]piperidine using diethyl sulfate. This achiral salt crystallizes in the chiral space group P21. One of the Si-C bonds in the cation is unusually long [1.9075 (12) Å], which correlates with the adjacent quaternary N+ atom and was verified by quantum chemical calculations. In the crystal, the components are linked by weak C-H⋯O hydrogen bonds: a Hirshfeld surface analysis was performed to further investigate these inter-molecular inter-actions and their effects on the crystal packing.

↵​Genetic Diversity and Population Structure of Two Faba Bean Mutant Populations Based on AFLP Markers

Background: Although induced mutagenesis can rapidly generate genetic diversity, every genotype responds differently to different mutagen treatments in induceing genetic diversity. This study assesses and compares the genetic diversity of two mutant faba bean populations. Methods: Two genotypes representing landrace (Hassawi 2) and inbred variety (ILB4347) were exposed to gamma radiation and diethyl sulfate (DES). Two hundred eighty-two samples derived from individual 140 M2 mutant plants of Hassawi 2 and ILB 4347 and two parental lines were characterized using 11 Amplified Fragment Length Polymorphism (AFLP) primer combinations. Result: 89,820 bands within 2,083 polymorphic alleles were generated from the samples. Genetic diversity comparison of the mutant populations revealed that each genotype had varying responses to different treatments. The two genotypes had a relatively similar response to gamma radiation, while different responses were recorded in DES-derived mutant plants. Based on the Nei’s genetic distance, the populations were separated based on the genotypic origin. The population structure analyses showed that the Hassawi 2 and ILB4347 mutant populations were clustered into three and two groups, respectively. The difference in the number of clusters between the two mutant populations explains their breeding history. The present diversity considered as a valuable material used for breeding and conservation purposes.

Mutation, Chemoprofiling, Dereplication, and Isolation of Natural Products from Penicillium oxalicum.

Diethyl sulfate (DES)-based chemical mutagenesis was applied on different fungal strains with the aim of diversifying the secondary metabolites. The mutant strain (VRE-MT1) of Penicillium oxalicum was subjected to dereplication (LCMS-based) and isolation of natural products, resulting in obtaining 10 molecules of bioactive potential. Metabolites, viz. tuckolide, methylpenicinoline, 2-acetyl-3,5-dihydroxy-4,6-dimethylbenzeneacetic acid, penicillixanthone A, brefeldin A 7-ketone, and antibiotic FD 549, were observed for the first time from P. oxalicum. The results of antimicrobial activity reveal that the compounds N-[2-(4-hydroxyphenyl)ethenyl]formamide, methylpenicinoline, and penipanoid A have potent antibacterial activity against Bacillus subtilis (ATCC 6633) with minimum inhibitory concentration (MIC) values of 16, 64, and 16 μM, respectively, and the compounds N-[2-(4-hydroxyphenyl)ethenyl]formamide, methylpenicinoline, and penipanoid A were found active against Escherichia coli (ATCC 25922), with MIC values of 16, 64, and 16 μM, respectively. Also, the metabolites N-[2-(4-hydroxyphenyl)ethenyl]formamide and tuckolide showed effective antioxidant activity in 2,2-diphenyl-1-picrylhydrazyl and 2,2′-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid scavenging assays. The mutant VRE-MT1 was found to have 8.34 times higher quantity of N-[2-(4-hydroxyphenyl)ethenyl]formamide as compared to the mother strain. The DES-based mutagenesis strategy has been found to be a potent tool to diversify the secondary metabolites in fungi.