Artemisia herba-alba sesquiterpenes: in silico inhibition in the ATP-binding pocket.

To identify antimicrobial leads for medical applications, metabolites from the aerial part of Artemisia herba-alba were extracted and chromatographically purified. Two new sesquiterpenes, 1ß,8α-dihydroxyeudesm-4-en-6ß,7α,11ßH-12,6-olide (1) and 1ß,6α,8α-trihydroxy, 11α-methyl-eudesma-4(15)-en-13-propanoate (2) along with a known eudesmanolide 11-epi-artapshin (3) were identified. Structures were determined by spectroscopic methods including 1D- and 2D-NMR as well as mass spectroscopy. Compound 3 inhibited Gram-positive bacteria Bacillus subtilis, Lactobacillus cereus and Staphylococcus aureus and exhibited antifungal activity against the pathogenic fungus F. solani. The mode-of-action of these antimicrobial sesquiterpenes as bacterial type II DNA topoisomerase and/or DNA gyrase B inhibitors were examined via in silico studies. Such molecular-docking studies were also employed to examine antifungal activity against an N-myristoyl transferase (NMT) target. Compound 3 had the greatest gyrase B binding affinity in the ATP-binding pocket and was found to possess an inhibitory action against non-invasive micro-test technology (NMT).

Statically controlled mycogenic-synthesis of novel biologically active silver-nanoparticles using Hafr Al-Batin desert truffles and its antimicrobial efficacy against pathogens.

In our search for new unconventional green-reducing agent, can be applied for biosynthesis of biologically active silver-nanoparticles, fruiting bodies (Ascocarps) of Truffle Terminia sp. were collected from the sandy desert of Hafr Al-Batin, Eastern Region, Saudi Arabia. The desert truffle showed the ability to reduce AgNO3 to Ag0 depending on their high content of proteins (1.74 mg/ml) in the aqueous extract of 30 mg/ml (w/v). The response surface methodology (RSM) with 13 experiments of 2-Factors-5-Levels central composite design was applied for controlling all possible combinations of AgNO3 concentrations and pH values of reaction mixture, which directly affect the particles morphology, size and biological activity. The antimicrobial effectiveness of all synthesized nanoparticles was evaluated against the pathogenic strains by agar diffusion method. The pathogenic Gram-positive Staphylococcus aureus, Bacillus subtilis, Lactobacillus cereus, and Gram-negative Escherichia coli, Salmonella enterica, yeast strain Candida albicans and the fungus Aspergillus niger were evaluated. The biologically active Truffle-AgNPs were characterized by UV-visible spectrophotometry, transmission electron microscopy (TEM), spectrum and dynamic light scatter (DLS), and Fourier Transformed Infrared (FTIR). Results obtained indicated that, the statistically controlled Truffle-AgNPs have great inhibitory role affecting different pathogenic strains, which gained much attention towards application of Hafr Al-Batin-Truffle as reducing and stabilizing biomaterial for green nano-drugs biosynthesis, to resist harmful pathogens threaten human health.

New antimicrobial metabolites from the medicinal herb Artemisia herba-Alba.

Artemisia herba-alba is widely used in traditional medicines for the treatment of several diseases. From the aerial parts organic extract of A. herba-alba, two new compounds, 1,3,8-trihydroxyeudesm-4-en-7α,11ßH-12,6α-olide (1) and 5-ß-​D-​glucopyranosyloxy​-​7-methoxy-​6H-​benzopyran-​2-​one (2), respectively, together with five known metabolites: 3α,8ß-dihydroxygermacr-4(15),9(10)-dien-7ß,11αH,12,6α-olide (3), 1ß,8α-dihydroxy-11α,13-dihydrobalchanin (4), 11-epiartapshin (5), tomenin (6) and benzoic acid, p-​(ß-​D-​glucopyranosyloxy)​-​methyl ester (7), were isolated and identified. The chemical structures were proven depending upon spectroscopic analysis, including 1 D/2D NMR as and ESI-MS. Compound 1 inhibited Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus. Compounds 2 and 3 exhibited antibacterial activity against both gram-positive and gram-negative bacteria.

Thermodynamics enhancement of Alternaria tenuissima KM651985 laccase by covalent coupling to polysaccharides and its applications.

This study full filed in enhancement of catalytic, thermodynamics and storage stability of Alternaria tenuissima KM651985 laccase by conjugation to sodium periodate oxidized starch. The starch conjugated A. tenuissima KM651985 laccase was active over a wide range of temperatures and pHs with the highest activity at 60 °C and 4, respectively. The thermal stability of conjugated A. tenuissima KM651985 laccase was indicated by, high T1/2 values (half life) 1076.16, 382.42 and 191.23 min at 50, 60 and 70 °C, respectively, low Kd (denaturation rate constant) 6.44 × 10-4, 18.13 × 10-4 and 36.25 × 10-4 min-1 at the same temperatures, high D-values (decimal reduction time) 3575.56, 1270.61 and 635.38 min at the same temperatures. Also, the thermal stability of conjugated A. tenuissima KM651985 laccase was emphasized by high ΔHd (enthalpy), high ΔGd (free energy) and low ΔSd (entropy). The conjugated A. tenuissima KM651985 laccase showed high effectiveness in dyes decolorization of Remazol Brilliant Blue R (RBBR) and Malachite Green (MG). Moreover, the addition of conjugated A. tenuissima KM651985 laccase with hemicellulolytic enzymes cocktail improved the saccharification of corn cobs, rice straw, corn cobs leaves and water hyacinth with the highest reducing sugar production 847.44 ±â€¯19.17 mg from corn cops.

In-situ green myco-synthesis of silver nanoparticles onto cotton fabrics for broad spectrum antimicrobial activity.

In the realm of green synthesis of metals nanoparticles for medical textile application, silver nanoparticles (AgNPs) were biosynthesized in situ cotton fabrics for the first time by using fungi for rendering cotton fabrics antimicrobial activity with abroad range towards different pathogenic organisms. Herein, five different isolated fungi from medicinal plants were identified and optimized their growth media prior examined their ability to reduce Ag+ ions to AgNPs in-situ cotton fabrics along with ex-situ method. Synthesis of AgNPs were characterized by making use of instruments e.g. UV-vis spectroscopy, Transmission Electron Microscopy (TEM), Selected Area Electron Diffraction (SAED), Scanning Electron Microscopy (SEM), and Fourier Transform Infrared (FTIR). Whereas antimicrobial activities of the resultant cotton fabrics were investigated against Gram positive (S. aureus ATCC29213), Gram negative (E. coli ATCC 25922), Yeast (C. albicans ATCC10321) and, fungi (A. niger NRC 53). Results revealed the successful biosynthesis of AgNPs using different fungus strains whether in-situ cotton fabrics or ex-situ manner. The size of the resultant AgNPs by ex-situ method were varied (5-20 nm). The antimicrobial activity of the in-situ treated cotton samples exhibited different behaviors towards both pathogenic bacteria and fungi. This manner opens up a new way to discover the ability of nanobiotechnology to provide world with substitutional aids mimic to synthetic materials.

Improvement of catalytic, thermodynamics and antifungal activity of constitutive Trichoderma longibrachiatum KT693225 exochitinase by covalent coupling to oxidized polysaccharides.

Our study full filled in two main goals preparation of constitutive exochitinase with low cost, utilizing non-chitin containing agricultural wastes, and improving the thermodynamics of purified Trichoderma longibrachiatum KT693225 exochitinase by covalent coupling to sodium periodate activated agar. Central composite design (CCD) was used to improve the chemical modification of Trichoderma longibrachiatum KT693225 exochitinase. Optimum temperature for conjugated exochitinase 60 °C was higher than native form 40 °C. Covalent coupling to oxidized agar caused 4.32, 2.75 and 2.44-fold increase in half-life values at 50, 55 and 60 °C, respectively. Also, conjugated exochitinase showed higher D-values (decimal reduction time) 1790.49 compared to 733.08 min for native form at 60 °C. Moreover, conjugated form had lower deactivation constant rate (kd) 0.39 × 10-3 min-1at 60 °C than native form 1.7 × 10-3 min-1. Native exochitinase exhibited higher activation energy (Ea) 3.39 Kcal·mol-1 and lower energy for denaturation (Ed) 6.88 Kcal·mol-1 compared to 3.21 and 13.05 Kcal·mol-1, respectively for conjugated form. The values of thermodynamic parameters for inactivation of native and conjugated exochitinase indicated that conjugation significantly decreased entropy (ΔS°) and increased enthalpy (ΔH°) and free energy (ΔG°) of deactivation. Conjugated exochitinase exhibited higher antifungal effect against Alternaria alternata, Fusarium oxysporium and Aspergillus niger than native form.

Purification, physicochemical and thermodynamic studies of antifungal chitinase with production of bioactive chitosan-oligosaccharide from newly isolated Aspergillus griseoaurantiacus KX010988.

In our search for chitinase and chitosanase producer from unconventional sources, the marine-derived fungus Aspergillus griseoaurantiacus KX010988 was obviously the best producer of the highest chitinase and chitosanase activities by solid state fermentation of potato shells. Chitinase was purified in three steps involving ammonium sulphate precipitation, DEAE-cellulose ion-exchange chromatography and Sephacryl S-300 gel chromatography. 12.55 fold increase in purity with a recovery of 17.6 was obtained. The molecular mass of the purified chitinase was found to be 130kDa. It was optimally active at pH 4.5 and 40°C. Km and Vmax values were 0.22mgmL-1 and 19.6µmolemin-1mg-1 respectively. Mn2+ and Zn2+ ions lead to increased chitinase activity. While Fe2+and Cu2+ions strongly inhibited the chitinase activity. The thermodynamics of pure chitinase including activation energy for thermal denaturation (Ea,d), change of free energy (ΔGd), enthalpy(ΔHd), entropy(ΔSd) and half life values (T1/2) at 40, 50 and 60°C were determined. Chitinase showed antifungal activity against pathogenic fungus Fusarium solani. Chitosanase was partially purified by acetone precipitation (50-75%) v/v concentration. The hydrolytic products of moderate molecular weight of chitosan by chitosanase were analyzed by thin layer chromatography (TLC) after 12 and 24h respectively. Chitosan-oligosaccharides showed good antibacterial and antioxidant activities.

3-Oxo-γ-costic acid fungal-transformation generates eudesmane sesquiterpenes with in vitro tumor-inhibitory activity.

While select eudesmane sesquiterpenes exhibit anti-neoplastic activity, tumor-inhibition for costic-acids has not been established. Here biological activity of 3-oxo-γ-costic acid (1), previously isolated from Chiliadenus montanus, as well as new sesquiterpenes (2-5) and the known derivative, 3-oxoeudesma-1,4,11(13)-trien-7-1061αH-l2-oic acid (6), all produced from 1 by the fungus Athelia rolfsii, are reported. Structures were elucidated using MS and NMR spectroscopy with activity-screening utilizing human colon- and lung-tumor lines, Caco-2 and A549 respectively. Compound 1 exhibited anti-proliferative activity against Caco-2 (IC50 39µM) and 2 was active against A549 (IC50 74µM) suggesting therapeutic potential for the original substrate and a bio-transformed product.

Antimicrobial sesquiterpene lactones from Artemisia sieberi.

Two new sesquiterpene lactones 3R, 8R-dihydroxygermacr-4(15),9(10)-dien-6S,7S,11RH,12,6-olide (1) and 1R, 8S-dihydroxy-11R,13-dihydrobalchanin(2), together with two known compounds 11-epiartapshin (3) and 3'-hydroxygenkwanin (4), were isolated from Artemisia sieberi. Their structures were elucidated by 1D, 2D NMR, MS, and X-ray diffraction. Compound 4 inhibited Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus with Minimal inhibitory concentration values of 50 and 25 µg/disk, respectively. All the isolated compounds exhibited moderate antifungal activities.

Covalent immobilization of microbial naringinase using novel thermally stable biopolymer for hydrolysis of naringin.

Naringinase induced from the fermented broth of marine-derived fungus Aspergillus niger was immobilized into grafted gel beads, to obtain biocatalytically active beads. The support for enzyme immobilization was characterized by ART-FTIR and TGA techniques. TGA revealed a significant improvement in the grafted gel's thermal stability from 200 to 300 °C. Optimization of the enzyme loading capacity increased gradually by 28-fold from 32 U/g gel to 899 U/g gel beads, retaining 99 % of the enzyme immobilization efficiency and 88 % of the immobilization yield. The immobilization process highly improved the enzyme's thermal stability from 50 to 70 °C, which is favored in food industries, and reusability test retained 100 % of the immobilized enzyme activity after 20 cycles. These results are very useful on the marketing and industrial levels.

Optimization of inulinase production from low cost substrates using Plackett-Burman and Taguchi methods.

Four marine-derived fungal isolates were screened for the production of inulinase enzyme from low cost substrates under solid state fermentation (SSF), one of them identified as Aspergillus terreus showed the highest inulinase activity using artichoke leaves as a solid substrate. Sequential optimization strategy, based on statistical experimental designs was employed to optimize the composition of the medium, including Plackett-Burman and Taguchi's (L9 3(4)) orthogonal array designs. Under the optimized conditions, inulinase activity (21.058 U/gds) reached the predicted maximum activity derived from the taguchi methodology, which increased about 4.79-folds the initial production medium. Fructose was produced, as an end product of inulin hydrolysis proving that the enzyme produced was exoinulinase. The marine-derived A. terreus is suggested as a new potential candidate for industrial enzymatic production of fructose from low cost substrate containing inulin as an economic source.