Ipsita K. Sen

Green synthesis of gold nanoparticles using a glucan of an edible mushroom and study of catalytic activity

Gold nanoparticles were synthesized by reducing chloroauric acid with a glucan, isolated from an edible mushroom Pleurotus florida, cultivar Assam Florida. Here, glucan acts as reducing as well as stabilizing agent. The synthesized gold nanoparticles were characterized by UV-visible spectroscopy, HR-TEM, XRD, SEM, and FT-IR analysis. The results indicated that the size distribution of gold nanoparticles (Au NPs) changed with the change in concentration of chloroauric acid (HAuCl(4)). The resulting Au NPs-glucan bioconjugates function as an efficient heterogeneous catalyst in the reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP), in the presence of sodium borohydride. The reduction of 4-nitrophenol with Au NPs-glucan bioconjugates followed pseudo-first-order kinetics. The effect of particle size and gold loading on reduction rate of 4-NP was studied with Au NPs-glucan bioconjugates prepared with different concentrations of HAuCl(4). The synthesis of catalytically active Au NPs using a pure mushroom polysaccharide of known structure is reported for the first time.

Green synthesis of silver nanoparticles using glucan from mushroom and study of antibacterial activity

This work demonstrates synthesis of silver nanoparticles (AgNPs) using glucan isolated from a mushroom Pleurotus florida blue variant. UV-vis spectroscopy showed maximum absorbance at 425 nm due to surface plasmon resonance of AgNPs. Average diameter of the synthesized AgNPs was 2.445 ± 1.08 nm as revealed from TEM analysis. XRD analysis confirmed the face-centered cubic (fcc) crystalline structure of metallic silver. The synthesized AgNPs-glucan conjugates exhibited antibacterial activity against multiple antibiotic resistant (MAR) bacterium Klebsiella pneumoniae YSI6A and the activity was possibly due to damage of cellular macromolecules by the generation of reactive oxygen species (ROS) which was supported by observed degradation of bacterial DNA. Decrease of bactericidal effect of AgNPs-glucan conjugates in dose-dependent manner in presence of a ROS scavenger histidine further ascertained the involvement of ROS in antibacterial activity. AgNPs-glucan conjugates at LD50 dose caused least damage (0.68% hemolysis) to human RBCs. This particular dose of AgNPs-glucan conjugates in combination with each of the four antibiotics (ampicillin, azithromycin, cefepime and tetracycline) to which K. pneumoniae YSI6A was resistant, showed synergistic effect to inhibit almost 100% bacterial growth. It thus opens an avenue to use antibiotics in combination with minimum dosages of AgNPs-glucan conjugates to control MAR bacteria.

Antioxidant and immunostimulant β-glucan from edible mushroom Russula albonigra (Krombh.) Fr.

A water soluble β-glucan (PS) with an average molecular weight ∼1.95 × 10(5)Da was isolated from the alkaline extract of ectomycorrhizal edible mushroom, Russula albonigra (Krombh.) Fr. and found to consist of terminal, (1 → 3)-, (1 → 6)-, and (1 → 3,6)-linked β-D-glucopyranosyl moieties in a ratio of nearly 1:2:2:1. The structure of this PS was elucidated on the basis of total hydrolysis, methylation analysis, Smith degradation, partial hydrolysis, and 1D/2D NMR experiments. On the basis of these experiments, the repeating unit of the PS was found to contain a backbone of three (1 → 6)-β-D-glucopyranosyl residues, one of which was branched at O-3 position with the side chain consisting of two (1 → 3)-β-D-glucopyranosyl and a terminal β-D-glucopyranosyl residue. This PS showed in vitro macrophage activation by NO production as well as splenocytes and thymocytes proliferation. Moreover, it also exhibited potent antioxidant activities.

Structure elucidation and antioxidant properties of a soluble β-d-glucan from mushroom Entoloma lividoalbum

A water soluble branched β-D-glucan (PS-I) with an average molecular weight ~2.1×10(5) Da was isolated from alkaline extract of the fruit bodies of the edible mushroom Entoloma lividoalbum (Kühner & Romagn) Kubička which consists of terminal β-D-glucopyranosyl, (1→3)-β-D-glucopyranosyl, (1→6)-β-D-glucopyranosyl, and (1→3,6)-β-D-glucopyranosyl moieties in a molar ratio of nearly 1:3:2:1. The structure of PS-I was elucidated using acid hydrolysis, methylation analysis, periodate oxidation study, partial hydrolysis, and 1D/2D NMR experiments. The repeating unit of the polysaccharide (PS-I) contains a backbone chain of three (1→6)-β-D-glucopyranosyl residues, one of which was branched at O-3 position with the side chain consisting of three (1→3)-β-D-glucopyranosyl and a terminal β-D-glucopyranosyl residues. Total antioxidant capacity of 1mg PS-I was measured and found equivalent to 70±15 μg of ascorbic acid. The PS-I was found to possess hydroxyl and superoxide radical-scavenging activities with EC50 values of 480 and 150 μg/mL, respectively. The reducing power of PS-I was determined 0.5 at 480 μg/mL.

Glucan of a somatic hybrid mushroom, pfls1h: structural characterization and study of immunological activities

A water-soluble glucan (PS-I) was isolated from the aqueous extract of the fruit bodies of a hybrid mushroom, pfls1h of Pleurotus florida and Lentinus squarrosulus (Mont.) Singer. Structural characterization of PS-I was carried out using total hydrolysis, methylation analysis, periodate oxidation, and NMR experiments ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HSQC, and HMBC). Methylation analysis revealed that PS-I was composed of (1→3, 6), (1→3), (1→6)-linked and terminal β-d-glucopyranosyl residues in a relative proportion of approximately 1:1:1:1. The repeating unit of the glucan consists of a backbone chain of two (1→6)-β-d-glucopyranosyl residues, one of which is branched at O-3 position with (1→3)-β-d-glucopyranosyl and terminated with a β-d-glucopyranosyl residue. Study of immunological activity revealed that PS-I stimulates the splenocytes, thymocytes and macrophages.

Structural characterization and study of immunoenhancing properties of a glucan isolated from a hybrid mushroom of Pleurotus florida and Lentinula edodes

A water soluble glucan isolated from hot aqueous extract of fruit bodies of an edible hybrid mushroom Pfle1r of Pleurotus florida and Lentinula edodes showed macrophages, splenocytes, and thymocytes activation. The glucan consists of terminal, (1→3,6)-linked, and (1→6)-linked β-D-glucopyranosyl moieties in a molar ratio of nearly 1:1:3. On the basis of acid hydrolysis, methylation, periodate oxidation study, and NMR studies ((1)H, (13)C, DEPT-135, TOCSY, DQF-COSY, NOESY, ROESY, HSQC, and HMBC), the structure of the repeating unit of the glucan was established as: [structure: see text].

Glucan from hot aqueous extract of an ectomycorrhizal edible mushroom, Russula albonigra (Krombh.) Fr.: structural characterization and study of immunoenhancing properties

A water soluble glucan (PS-I) was isolated from the hot aqueous extract of the fruit bodies of an ectomycorrhizal edible mushroom, Russula albonigra (Krombh.) Fr. The total hydrolysis, methylation analysis, periodate oxidation, and NMR ((1)H, (13)C, DEPT-135, TOCSY, DQF-COSY, NOESY, ROESY, HSQC, and HMBC) studies revealed the presence of the following repeating unit in the polysaccharide: This glucan showed excellent activation of macrophages as well as splenocytes and thymocytes in vitro.

Partial characterization and flocculating behavior of an exopolysaccharide produced in nutrient-poor medium by a facultative oligotroph Klebsiella sp. PB12.

A facultative oligotrophic strain from the water sample of River Mahananda, Siliguri India was selected for its property to produce exopolysaccharide (EPS) in nutrient-poor (oligotrophic) medium. Viability assay of the strain was performed in sterile liquid LB, R2A, river water and diluted (10(-3)) LB at 30°C and pH 7 to understand oligotrophy. The selected strain was identified by 16S rRNA gene sequencing and designated as Klebsiella sp. PB12. Phylogenetic analysis showed its closest relationship with Klebsiella variicola ATCC BAA-830(T). Purification of EPS was performed by ethanol precipitation, dialysis and freeze-drying. Chemical analysis revealed that purified EPS was mainly composed of 72.32% (w/w) neutral sugar and 14.12% (w/w) uronic acids. Fourier transform infrared (FT-IR) spectroscopy indicated the presence of hydroxyl, carboxylic and methoxyl functional groups. The optimal dosages for flocculation of activated carbon suspension were 17 mg/l EPS and 4 mM CaCl(2). EPS showed flocculating rate of above 80% over a wide range of pH (pH 3-10) whereas, more than 90% rate was noted in the temperature range (10-50°C) tested in presence of CaCl(2). Moreover, EPS showed characteristic emulsifying activity with toluene (66.6%), n-hexadecane (65%), olive oil (63.3%) and kerosene (50%). The apparent molecular weight of the EPS was ~2 × 10(5) Da.

Structural and immunological studies of hetero polysaccharide isolated from the alkaline extract of Tricholoma crassum (Berk.) Sacc.

An immunoenhancing water-soluble hetero polysaccharide was isolated from an alkaline extract of the fruit bodies of an ectomycorrhizal edible mushroom, Tricholoma crassum (Berk.) Sacc. The structure of the molecule was investigated using acid hydrolysis, methylation analysis; periodate oxidation study, and NMR spectroscopy techniques ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HSQC, and HMBC). On the basis of the above-mentioned experiments the structure of the repeating unit of the polysaccharide was established as: This polysaccharide exhibited splenocyte, thymocyte as well as macrophage activations.

A glucan from an ectomycorrhizal edible mushroom Tricholoma crassum (Berk.) Sacc.: isolation, characterization, and biological studies

A water soluble glucan of average molecular weight ∼1.74×10(5)Da was isolated from hot aqueous extract of the fruiting bodies of an ectomycorrhizal edible mushroom Tricholoma crassum (Berk.) Sacc. The structure of this glucan was elucidated on the basis of total hydrolysis, methylation analysis, Smith degradation, and 1D/2D NMR studies. Based on the above experiments the repeating unit of the glucan was established as: [formula see text]. This glucan showed macrophage activation in vitro by NO production in a dose dependent manner and strong splenocyte and thymocyte immunostimulation in mouse cell culture medium. It also exhibited good inhibition activity toward lipid peroxidation.