Ultrafiltration and a series of chromatographic steps were used to isolate and purify polysaccharides from Tremella aurantialba fruit bodies.Three crude fractions(TAP50w,TAP10-50w,and TAP1-10w),five semi-purified frac...Ultrafiltration and a series of chromatographic steps were used to isolate and purify polysaccharides from Tremella aurantialba fruit bodies.Three crude fractions(TAP50w,TAP10-50w,and TAP1-10w),five semi-purified fractions(TAPA-TAPE),and one purified fraction(TAPA1) were obtained.A sulfated derivative of TAPA1(TAPA1-s) was prepared by chemical modification.The immunostimulating activity of the polysaccharide fractions in vitro was determined using the mouse spleen lymphocyte proliferation assay.Of the three crude fractions tested,cell prolifera-tion rates were increased most by TAP50w.Furthermore,TAPA1-s was markedly more stimulatory than TAPA1,indicating that sulfonation was an effective way to enhance the immunostimulating activity of polysaccharide.展开更多
A yeast-like fungus strain B1 isolated from wild fungus Tremella aurantialba was identified and initially characterized. Two phylogenetic trees were generated based on the sequences of large subunit ribosomal RNA gene...A yeast-like fungus strain B1 isolated from wild fungus Tremella aurantialba was identified and initially characterized. Two phylogenetic trees were generated based on the sequences of large subunit ribosomal RNA gene D1/D2 regions and internal transcribed spacer (ITS) regions of related fungi, respectively. The analysis of D1/D2 regions and ITS sequences showed that fungus B1 was clustered together with T. aurantialba, T. aurantia and T. microspore in the phylogenetic trees. Both the morphological characteristic and phylogenetic analysis established that fungus B1 was one of the anamorph strains of T. aurantialba and belongs to Tremella genus. A fermentation medium for exopolysaccharides (EPS) production by T. aurantialba B1 . Plackett-Burmen design was used to evaluate the effects of different components in the culture medium. Glucose and yeast extract have significant influence on the EPS production. The concentrations of two factors were optimized subsequently using central composite design and response surface analysis. The results showed that 49.2 g/L glucose and 10.4 g/L yeast extract could lead to the maximum production of EPS (4.99 g/L). The optimized medium led to a 1.5-fold enhancement of the production of EPS by T. aurantialba B1 , as compared with that without optimization.展开更多
[Objective] This study aimed to explore the protoplast preparation and re- generation conditions of Tremella aurantialba. [Method] Optimal combination of five factors affecting the protoplast preparation and regenerat...[Objective] This study aimed to explore the protoplast preparation and re- generation conditions of Tremella aurantialba. [Method] Optimal combination of five factors affecting the protoplast preparation and regeneration of Tremella aurantialba was selected by using orthogonal experiment, including enzyme system, culture age, enzymolysis temperature, enzymolysis duration and osmotic stabilizer. [Result] The results showed that there were three release modes of Tremella aurantialba proto- plasts： release from top in the early period of enzymolysis, release from the side and release in situ. The optimal protoplast preparation condition was selecting mycelium cultured in liquid medium for 3 d, for enzymolysis in mixed enzyme solu- tion containing 1% of cellulase ＋1% of snailase ＋1% of lywallzyme at 36 ℃; for 4 h, with 0.6 mol/L sucrose as osmotic stabilizer, and the obtained preparation rate had reached 1.76 ×10^7 protoplasts/ml. The optimal regeneration condition was using mycelium cultured in liquid medium for 5 d, for enzymolysis in enzyme solution con- taining 1.5% of lywallzyme at 33 ℃ for 3 h, with 0.6 mol/L sucrose as osmotic sta- bilizer, and the regeneration rate had reached 0.22%. [Conclusion] This study provid- ed valuable support for protoplast fusion, ultraviolet mutation breeding and genetic engineering research.展开更多
基金Project (No.2006BAD06B08) supported by the National Key Technology R & D Program of China
文摘Ultrafiltration and a series of chromatographic steps were used to isolate and purify polysaccharides from Tremella aurantialba fruit bodies.Three crude fractions(TAP50w,TAP10-50w,and TAP1-10w),five semi-purified fractions(TAPA-TAPE),and one purified fraction(TAPA1) were obtained.A sulfated derivative of TAPA1(TAPA1-s) was prepared by chemical modification.The immunostimulating activity of the polysaccharide fractions in vitro was determined using the mouse spleen lymphocyte proliferation assay.Of the three crude fractions tested,cell prolifera-tion rates were increased most by TAP50w.Furthermore,TAPA1-s was markedly more stimulatory than TAPA1,indicating that sulfonation was an effective way to enhance the immunostimulating activity of polysaccharide.
基金Supported by the Key Project of National 9th Five-Year Plan Program (No.96-C02-03-06)
文摘A yeast-like fungus strain B1 isolated from wild fungus Tremella aurantialba was identified and initially characterized. Two phylogenetic trees were generated based on the sequences of large subunit ribosomal RNA gene D1/D2 regions and internal transcribed spacer (ITS) regions of related fungi, respectively. The analysis of D1/D2 regions and ITS sequences showed that fungus B1 was clustered together with T. aurantialba, T. aurantia and T. microspore in the phylogenetic trees. Both the morphological characteristic and phylogenetic analysis established that fungus B1 was one of the anamorph strains of T. aurantialba and belongs to Tremella genus. A fermentation medium for exopolysaccharides (EPS) production by T. aurantialba B1 . Plackett-Burmen design was used to evaluate the effects of different components in the culture medium. Glucose and yeast extract have significant influence on the EPS production. The concentrations of two factors were optimized subsequently using central composite design and response surface analysis. The results showed that 49.2 g/L glucose and 10.4 g/L yeast extract could lead to the maximum production of EPS (4.99 g/L). The optimized medium led to a 1.5-fold enhancement of the production of EPS by T. aurantialba B1 , as compared with that without optimization.
文摘[Objective] This study aimed to explore the protoplast preparation and re- generation conditions of Tremella aurantialba. [Method] Optimal combination of five factors affecting the protoplast preparation and regeneration of Tremella aurantialba was selected by using orthogonal experiment, including enzyme system, culture age, enzymolysis temperature, enzymolysis duration and osmotic stabilizer. [Result] The results showed that there were three release modes of Tremella aurantialba proto- plasts： release from top in the early period of enzymolysis, release from the side and release in situ. The optimal protoplast preparation condition was selecting mycelium cultured in liquid medium for 3 d, for enzymolysis in mixed enzyme solu- tion containing 1% of cellulase ＋1% of snailase ＋1% of lywallzyme at 36 ℃; for 4 h, with 0.6 mol/L sucrose as osmotic stabilizer, and the obtained preparation rate had reached 1.76 ×10^7 protoplasts/ml. The optimal regeneration condition was using mycelium cultured in liquid medium for 5 d, for enzymolysis in enzyme solution con- taining 1.5% of lywallzyme at 33 ℃ for 3 h, with 0.6 mol/L sucrose as osmotic sta- bilizer, and the regeneration rate had reached 0.22%. [Conclusion] This study provid- ed valuable support for protoplast fusion, ultraviolet mutation breeding and genetic engineering research.
