Green synthesis of silver nanoparticles using Phlebopus portentosus polysaccharide and their antioxidant, antidiabetic, anticancer, and antimicrobial activities.

Silver nanoparticles (AgNPs) by green synthesis from fungi polysaccharides are attracting increasing attention owing to their distinctive features and special applications in numerous fields. In this study, a cost-effective and environmentally friendly biosynthesizing AgNPs method with no toxic chemicals involved from the fruiting body polysaccharide of Phlebopus portentosus (PPP) was established and optimized by single factor experiment and response surface methodology. The optimum synthesis conditions of polysaccharide-AgNPs (PPP-AgNPs) were identified to be the reaction time of 140 min, reaction temperature of 94 °C, and the PPP: AgNO3 ratio of 1:11.5. Formation of PPP-AgNPs was indicated by visual detection of colour change from yellowish to yellowish brown. PPP-AgNPs were characterized by different methods and further evaluated for biological activities. That the Ultraviolet-visible (UV-Vis.) spectroscopy displayed a sharp absorption peak at 420 nm confirmed the formation of AgNPs. Fourier transform infrared (FTIR) analysis detected the presence of various functional groups. The lattice indices of (111), (200), (220), and (331), which indicated a faced-centered-cubic of the Ag crystal structure of PPP-AgNPs, was confirmed by X-ray diffraction (XRD) and the particles were found to be spherical through high resolution transmission electron microscopy (HRTEM). Energy dispersive X-ray spectroscopy (EDS) determined the presence of silver in PPP-AgNPs. The percentage relative composition of elements was determined as silver (Ag) 82.5 % and oxygen (O) 17.5 % for PPP-AgNPs, and did not exhibit any nitrogen peaks. The specific surface area of PPP-AgNPs was calculated to be 0.5750 m2/g with an average pore size of 24.33 nm by BET analysis. The zeta potential was -4.32 mV, which confirmed the stability and an average particle size of 64.5 nm was calculated through dynamic light scattering (DLS). PPP-AgNPs exhibited significant free radical scavenging activity against DPPH with an IC50 value of 0.1082 mg/mL. The MIC values of PPP-AgNPs for E. coli, S. aureus, C. albicans, C. glabrata, and C. parapsilosis are 0.05 mg/mL. The IC50 value of the inhibition of PPP-AgNPs against α-glucosidase was 11.1 µg/mL, while the IC50 values of PPP-AgNPs against HepG2 and MDA-MB-231 cell lines were calculated to be 14.36 ± 0.43 µg/mL and 40.05 ± 2.71 µg/mL, respectively. According to the evaluation, it can be concluded that these green-synthesized and eco-friendly PPP-AgNPs are helpful to improve therapeutics because of significant antioxidant, antimicrobial, antidiabetic, and anticancer properties to provide new possibilities for clinic applications.

Four New Species of Strobilomyces (Boletaceae, Boletales) from Hainan Island, Tropical China.

Strobilomyces, one of the most noticeable genera of Boletaceae (Boletales), is both ecologically and economically important. Although many studies have focused on Strobilomyces in China, the diversity still remains incompletely understood. In the present study, several collections of Strobilomyces from Hainan Island, tropical China were studied based on morphology and molecular phylogenetic analyses. Four species are described as new, viz. S. baozhengii, S. conicus, S. hainanensis, and S. pachycystidiatus. Detailed descriptions, color photos of fresh basidiomata, and line drawings of microstructures of the four species are presented.

Structural characterization, and in vitro hypoglycemic activity of a polysaccharide from the mushroom Cantharellus yunnanensis.

A polysaccharide CY-2 from C. yunnanensis was obtained through a process of consecutive water extraction, alcohol precipitation, and DEAE-52 fast-flow chromatography. CY-2, with an average molecular weight of 2.69 × 104 Da mainly consisted of glucose and mannose with a molar ratio of 33.5: 56.9. Infrared spectrum (IR), methylation analysis, and nuclear magnetic resonance (NMR) results revealed that CY-2 may have a backbone consisting of →6)-α-D-Manp-(1 â†’ 3)-ß-D-Glcp-(1→, and branch chain ß-D-Glcp-(1→. Meanwhile, CY-2 had a higher inhibition rate on α-glucosidase activity compared with other fractions (CY-0, CY-1, and CY-4) and was a mixed competitive inhibitor. In addition, CY-2 at the concentration of 10 µg/mL presented a superior power to improve glucose consumption and metabolism in HepG2 cells compared with metformin. Overall, these findings highlight the potential value of CY-2 as a hypoglycemic agent.

Extraction, Characterization, and In Vitro Hypoglycemic Activity of a Neutral Polysaccharide from the New Medicinal Mushroom Cantharellus yunnanensis (Agaricomycetes).

Polysaccharides serve as promising ingredients for health-beneficial functional foods, while there were no investigations into the structural characterizations and bioactivities of an edible mushroom Cantharellus yunnanensis. In the study, crude polysaccharides from this mushroom were extracted by hot water and isolated by ethanol precipitation. Then, a neutral polysaccharide (named CY-1) was purified from the crude polysaccharide by deproteinization with Sevag reagent, decolorization with a kind of macroporous adsorption resin SP-825, DEAE-52 cellulose column chromatography and dialysis. The physicochemical properties of CY-1 were characterized by UV, IR, SEM, NMR, and HPLC analyses. Structural characterizations revealed that CY-1 is a homogeneous heteropolysaccharide with an average molecular weight of 3.06 × 104 Da. CY-1 exhibited a honeycomb structure with an irregular branching shape, and it was composed of mannose, glucose, fucose, xylose, arabinose, galactose, rhamnose, and glucuronic acid, with molar fractions of 54.83%, 25.11%, 10.28% 4.53%, 2.12%, 1.64%, 0.83%, and 0.64%, respectively. In vitro hypoglycemic tests showed that CY-1 had an inhibitory effect on α-glucosidase. In addition, CY-1 of 160 µg/mL promoted glucose consumption in normal HepG2 cells. These results suggested that CY-1 may be a potential hypoglycemic agent.

Notes on four species of Russula subgenus Heterophyllidiae (Russulaceae, Russulales) from southern China.

Heterophyllidiae, one of the main subgenus of Russula (Russulaceae, Russulales), is both ecologically and economically important. Although many studies have focused on subgenus Heterophyllidiae in China, the diversity, taxonomy, and molecular phylogeny still remained incompletely understood. In the present study, two new species, R. discoidea and R. niveopicta, and two known taxa, R. xanthovirens and R. subatropurpurea, were described based on morphology and molecular phylogenetic analyses of ITS and 28S DNA sequences with new collections of subgenus Heterophyllidiae from southern China. Both morphological and phylogenetic analyses consistently confirmed that R. niveopicta and R. xanthovirens belong to the subsect. Virescentinae, R. discoidea and R. subatropurpurea come under subsect. Heterophyllae, and R. prasina is synonymized with R. xanthovirens.

Updated taxonomy of Chinese Cantharellus subgenera Afrocantharellus and Magni (Hydnaceae, Cantharellales): Three new taxa and amended descriptions of one previous species.

Cantharellus, one of the main genera of Hydnaceae (Cantharellales), is both ecologically and economically important. Although many studies have focused on this genus in China, the taxonomy should be further updated. In the present study, Cantharellus subgenera Afrocantharellus and Magni were investigated based on morphology and molecular phylogenetic analyses with new collections from China. Five phylogenetic species were recognized among the studied collections, three of which were described as new: C. bellus, C. cineraceus, and C. laevigatus; one was previously described taxon: C. hygrophoroides; and the remaining species was not defined due to the paucity of the materials. Among the four described species, both C. bellus and C. laevigatus are members of subgen. Magni, whereas C. cineraceus and C. hygrophoroides belong to subgen. Afrocantharellus.

Purification and Structure Characterization of the Crude Polysaccharide from the Fruiting Bodies of Butyriboletus pseudospeciosus and Its Modulation Effects on Gut Microbiota.

Polysaccharides from the species of Boletaceae (Boletales, Agaricomycetes, Basidiomycota) are economically significant to both functional foods and medicinal industries. The crude polysaccharide from Butyriboletus pseudospeciosus (BPP) was prepared, and its physicochemical properties were characterized through the use of consecutive experimental apparatus, and its impact on the gut microbiota of Kunming mice was evaluated. Analyses of the structure characteristics revealed that BPP was mainly composed of Man, Glc, and Gal, possessing the pyranose ring and ß/α-glycosidic linkages. TG analysis exhibited that BPP had great heat stability. The SEM observation performed demonstrated that BPP appeared with a rough, dense, and porous shape. Through the BPP intervention, the serum and fecal biochemical index in mice can be improved obviously (p < 0.05). The abundance of beneficial microbiota in the BPP-treated group was significantly increased, while the abundance of harmful microbiota was significantly decreased (p < 0.05). Based on the Tax4Fun, we also revealed the relationship between the species of gut microbiota and showed that the high dose of BPP has significantly changed the functional diversities compared with those in other groups (p < 0.05). The results suggest that B. pseudospeciosus could serve as potential functional food or medicine.

Preparation, structural characterisation, and antioxidant activities of polysaccharides from eight boletes (Boletales) in tropical China.

Polysaccharides in boletes (Boletales) are economically significant to both function food and medicinal industries. The polysaccharides were extracted from the fruit bodies of eight boletes, namely, Aureoboletus longicollis, Butyriboletus hainanensis, Crocinoboletus rufoaureus, Hemioporus japonicus, Neoboletus infuscatus, Neoboletus obscureumbrinus, Tylopilus otsuensis, Xanthoconium fusciceps, which were collected from tropical China; their physicochemical properties and antioxidant activities were characterised and evaluated, respectively. The results revealed that the polysaccharides among the eight boletes were mainly composed of glucose, mannose, and galactose, with a broad molecular weight range, and contained a pyranose ring revealed by FT-IR and NMR spectral analyses. Many factors such as different species of boletes, geographic conditions, molecular weight, configuration, and monosaccharide content may affect the antioxidant power of polysaccharides, simultaneously, instead of one single factor. The antioxidant activities of the polysaccharides were measured according to in vitro assays of DPPH scavenging, superoxide anion scavenging, and ferrous ion reducing tests. The polysaccharide of C. rufoaureus has greatly superior antioxidant activity and it could serve as potential functional food or medicine.

A Contribution to Knowledge of Craterellus (Hydnaceae, Cantharellales) in China: Three New Taxa and Amended Descriptions of Two Previous Species.

Species of Craterellus (Hydnaceae, Cantharellales) in China are investigated on the basis of morphological and molecular phylogenetic analyses of DNA sequences from nuc 28S rDNA D1-D2 domains (28S) and nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 region. Five species are recognized in China, of which three of them are described as new, viz. C. fulviceps, C. minor, and C. parvopullus, while two of them are previously described taxa, viz. C. aureus, and C. lutescens. A key to the known Chinese taxa of the genus is also provided.

Morphological and Phylogenetic Evidences Reveal Four New Species of Cantharellus Subgenus Cantharellus (Hydnaceae, Cantharellales) From China.

Species of Cantharellus subgenus Cantharellus are interesting and important for their mycorrhizal properties, medicinal values, and edibility. In China, there are many undescribed species of the subgenus. In this study, four new species of subg. Cantharellus, viz. Cantharellus albopileatus, Cantharellus chuiweifanii, Cantharellus pinetorus, and Cantharellus ravus from Hainan and Hunan Provinces, respectively, were described based on morphological and phylogenetic evidence as a contribution to the knowledge of the species diversity in China. Detailed descriptions, color photographs of fresh basidiomata, and line drawings of microstructures of these four new species are presented as well as comparisons with related species.

Rubroboletus flammeus (Boletaceae, Boletales), a novel species unveiled from subtropical China based on morphological and phylogenetic evidences.

Rubroboletus flammeus is described as a new species from subtropical China based on morphological and molecular phylogenetic analyses. It is morphologically characterized by a red to fiery-red basidioma with a dry pileus, a context in pileus white but that in stipe bright yellow, a stipe densely covered with spots, a blue color change of tissues, and a trichoderm-type pileipellis. Detailed descriptions, color photographs of fresh basidiomata, and line drawings of microscopic features of the new species are presented. A key to the known Chinese taxa of Rubroboletus is provided.

Neoboletus infuscatus, a new tropical bolete from Hainan, southern China.

Neoboletus infuscatus (Boletaceae, Boletales) is described as a new species from Yinggeling of Hainan Tropical Rainforest National Park, southern China. It is morphologically characterized by a large basidioma with a nearly glabrous, brownish yellow, yellowish brown to pale brown pileus, pores orangish red when young, yellowish brown to brown when old, context and hymenophore staining blue when injured, a yellow stipe with red punctuations, surfaces of the pileus and the stipe usually covered with a thin layer of white pruina when young. Phylogenetic analyses of DNA sequences from part of the 28S gene, the nuclear rDNA internal transcribed spacer (ITS) region, and part of the translation elongation factor 1-α gene (TEF1) also confirm that N. infuscatus forms an independent lineage within Neoboletus. Detailed morphological description, color photos of fresh basidiomata and line-drawings of microstructures are provided.

New and noteworthy boletes from subtropical and tropical China.

The morphology, ecology, and phylogenetic relationships of specimens of the family Boletaceae from subtropical and tropical China were investigated. Four species, Butyriboletushuangnianlaii, Lanmaoamacrocarpa, Neoboletusmultipunctatus, and Sutoriussubrufus, are new to science. Chalciporusradiatus and Caloboletusxiangtoushanensis are redescribed. Caloboletusguanyui is proposed to replace Boletusquercinus Hongo, an illegitimate later homonym. The recently described Tylopiluscallainus is synonymized with the Japanese Boletusvirescens, and the new combination T.virescens (Har. Takah. & Taneyama) N.K. Zeng et al. is proposed. Moreover, Neoboletus is treated as an independent genus based on evidence from morphology and molecular phylogenetic data in the present study, and many previously described taxa of Sutorius are recombined into Neoboletus: N.ferrugineus (G. Wu et al.) N.K. Zeng et al., N.flavidus (G. Wu & Zhu L. Yang) N.K. Zeng et al., N.hainanensis (T.H. Li & M. Zang) N.K. Zeng et al., N.obscureumbrinus (Hongo) N.K. Zeng et al., N.rubriporus (G. Wu & Zhu L. Yang) N.K. Zeng et al., N.sanguineoides (G. Wu & Zhu L. Yang) N.K. Zeng et al. , N.sanguineus (G. Wu & Zhu L. Yang) N.K. Zeng et al., and N.tomentulosus (M. Zang et al.) N.K. Zeng et al.

The genus Heimioporus in China.

Species of Heimioporus (Boletaceae, Boletales) in China are investigated on the basis of morphology and molecular phylogenetic analyses of DNA sequences from the nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 region and nuc 28S rDNA D1-D2 domains (28S) and translation elongation factor 1-α gene (TEF1). Three species are recognized in China: H. conicus, sp. nov., H. gaojiaocong, sp. nov., and the previously described H. japonicus. Heimioporus xerampelinus, originally described from southwestern China, is synonymized with H. japonicus; specimens from China previously regarded as H. retisporus and H. subretisporus are confirmed to represent H. japonicus and H. gaojiaocong, respectively. A key to the Chinese taxa of the genus is provided. A preliminary biogeographical analysis shows that Heimioporus in East Asia and Southeast Asia/Australia are closely related. Heimioporus japonicus is a geographically widespread species occurring in East Asia and Southeast Asia; however, no taxa common to East Asia and Australia were uncovered.

The genus Pulveroboletus (Boletaceae, Boletales) in China.

Species of Pulveroboletus (Boletaceae, Boletales) in China are investigated on the basis of morphology and molecular phylogenetic analyses of DNA sequences from nuc rDNA region encompassing the internal transcribed spacers 1 and 2, along with the 5.8S rDNA (ITS) and nuc 28S rDNA D1-D2 domains (28S) and sequences from the translation elongation factor 1-α gene (TEF1). Nine species are recognized in the country, three of them are described as new: P. flaviscabrosus, P. rubroscabrosus, and P. subrufus, five of them are previous described taxa: P. brunneopunctatus, P. brunneoscabrosus, P. macrosporus, P. reticulopileus, and P. sinensis, the remaining one is tentatively named P. cf. ridleyi, which will be further studied in the future. A key to the Chinese taxa of the genus is provided. Unexpectedly, the current study did not identify disjunct populations of Pulveroboletus species in East Asia and North/Central America.

A high throughput method for rapid screening of chitosanase-producing fungal strain under acidic conditions.

A novel high-throughput method was established for rapid screening of a large numbers of Aspergillus fumigatus (A. fumigatus) mutants with high chitosanase production under acidic culture condition by exploiting the fact that iodine can be used as the indicator to stain chitosan but is ineffective for chitooligosaccharides. The mutant population was generated by irradiating A. fumigatus CICC 2434 with Co(60)-γ rays. Mutants were cultured on acidic plates containing colloidal chitosan and preliminary screened according to diameter of haloes formed around colonies. Then, chitosanase production of the isolates were verified by dinitrosalicylic acid assay. Lastly, molecular masses on enzymolysis products of isolated mutants were rapidly compared by aniline blue plate assay. Using this method, the mutant strain Co-8 was selected, which had chitosanase activity of 24.87 U/mL (increased by 369.2 % as compared to that of its parental strain).Taking together, the method is easy, efficient and particularly suited to rapid screen acidophilic fungal strains with high chitosanase-production.

The genus Retiboletus in China.

Species of the genus Retiboletus (Boletaceae, Boletales) in China are investigated based on morphology and phylogenetic analyses of DNA sequences from nuc rDNA internal transcribed spacer (ITS) and partial 28S regions and sequences from the translation elongation factor 1-a gene (tef1a). Six lineages are recovered among the collections studied. Five of these are documented and presented in the present paper, including three new species and two new combinations. The remaining species is not described due to the paucity of material. The specimens from China identified as "R. ornatipes" or "R. retipes" are in fact R. sinensis or R. kauffmanii, those labeled "R. griseus" are either R. fuscus or R. pseudogriseus A key to all known taxa of the genus is provided. Phylogenetic relationships of taxa within Retiboletus are partially resolved. A preliminary biogeographical analysis shows that allied species of Retiboletus between eastern Asia and North/Central America are common but there are no Retiboletus species common to both continents. Species of Retiboletus in Japan and southern China are conspecific or closely related.

Purification and characterization of an intracellular α-glucosidase with high transglycosylation activity from A. niger M-1.

An intracellular α-glucosidase with high transglycosylation activity was purified from a mutant strain of Aspergillus niger M-1 by sequential chromatography using a DEAE-cellulose 52 column, a DEAE-Sepharose CL-6B column, and a Sephadex G-100 column. The molecular mass of the purified enzyme was determined to be 116 kD with no subunits and a pI of 5.23. Maximal α-glucosidase activity occurred at pH 6.0 and 50°C. The N-terminal amino acid sequences were identified as N-SVPGTEYVV-. The presence of Ca(2+) enhanced the enzyme activity by 20%, while the α-glucosidase activity was strongly inhibited by p-chloromercuribenzoate, N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride, monochloroacetic acid, and 2-mercaptoethanol. In addition, Ag(+), n-bromosuccinimide, and acetylacetone inhibited enzyme activity by 70%, 50%, and 22%, respectively. K(m) values of 4.32 m mol L(-1) and V(max) of 3.10 × 10(-2) mol L(-1) min(-1) were found for methyl-α-D-glucopyranoside (α-MG). Maltose was identified as the preferred substrate. The high-performance liquid chromatography (HPLC) analysis indicated that the oligosaccharide products contained 10.54% of isomaltose, 8.08% of panose, and 9.29% of isomaltotriose, and the amount of glucose, maltose, maltotriose, and maltotetrose was dropped from 22.21% to 15.80% using the purified enzyme in the solution of 25% maltose and 3% glucose. This intracellular α-glucosidase has potential applications in the synthesis of sugar derivatives and the investigation of associated mechanisms.

A high-throughput method for screening of Aspergillus niger mutants with high transglycosylation activity by detecting non-fermentable reducing sugar.

A novel high-throughput method was established for rapid screening of large numbers of Aspergillus niger mutants with high transglucosylation activity by exploiting that yeast can hardly hydrolyze isomaltooligosaccharides (IMO). Supernatants of A. niger fermentation were incubated with Saccharomyces cerevisiae to remove glucose and maltose, and the remaining non-reducing sugars, which is positively correlated with the amount of IMO, the products of transglucosylation reaction, were used as indicator of transglucosidase activity of A. niger and examined by dinitrosalicylic acid assay. Using this method, 15 stains that could convert liquefied cassava starch to IMO more efficiently were selected from 8721 A. niger mutants. Among them, mutant C-6181 strain had transglycosidase activity of 4.61 U/ml (increased by 122% compared to its parental strain) and IMO yield of 83.7%. Taking together, the method is easy, simple, efficient and cheap, and has great application potential in selection of transglucosidase-producing strains used in industrial IMO fermentation.

[Screening of microorganism producing polyunsaturated fatty acids with sugarcane molasses].

A strain name LB1 producing polyunsaturated fatty acids with sugarcane molasses was screened form soil samples by the methods of Sudan Black B stain and the determination of PUFAs' iodine value. Based on the grass morphological and physiological characteristics and the sequence similarity of ribosomal DNA-ITS, the strain was identified as Mucor recurvus sp.. The results of L64 (4(3)) orthogonal experiments indicated when Mucor recurvus was cultivated at 28 degrees C for 5 days with 160 r/min, the yields of PUFAs with 10(0) BX sugarcane molasses (pH 6.0) were up to 80.49% content of total fatty acids. The total lipids content was 57.08% of fermentation biomass, which were composed of 15.42% Oleic acid, 14.38% Linolenic, 23.55% gamma-Linolenic, 3.06% alpha-Linolenic, 9.87% Arachidonic acid, 8.14% Eicosapentaenoic acid, 6.07% Docosahexaenoic acid, etc.