[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
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
Extracellular polysaccharide from Tremella aurantialba (TEP) significantly inhibited H2O2-induced haemolysis of RBCs, significantly decreased the amount of malondialdehyde (MDA) produced in mice liver homogenates as a...Extracellular polysaccharide from Tremella aurantialba (TEP) significantly inhibited H2O2-induced haemolysis of RBCs, significantly decreased the amount of malondialdehyde (MDA) produced in mice liver homogenates as a result of autoxidation, and inhibited VitC-Fe2+-induced swelling of liver mitochondria.展开更多
以甘肃省兰州地区培育的金耳子实体为原料,经分离纯化得到分子质量(M_(w)=1.54×10^(6)Da)分布均一的金耳纯多糖(Tremella aurantialba pure polysaccharide,TAPP),其主要单糖组成为甘露糖∶葡萄糖醛酸∶葡萄糖∶木糖=55.20∶9.76∶...以甘肃省兰州地区培育的金耳子实体为原料,经分离纯化得到分子质量(M_(w)=1.54×10^(6)Da)分布均一的金耳纯多糖(Tremella aurantialba pure polysaccharide,TAPP),其主要单糖组成为甘露糖∶葡萄糖醛酸∶葡萄糖∶木糖=55.20∶9.76∶14.81∶20.23,含有少量半乳糖和半乳糖醛酸。傅里叶变换红外光谱、刚果红实验、扫描电子显微镜分析和热重分析结果表明,TAPP含有β-糖苷键和螺旋结构,表面微观形貌呈片状无规则聚集状态,170℃以下热稳定性良好。活性分析结果显示,TAPP对α-淀粉酶的和α-葡萄糖苷酶的抑制与浓度呈正相关,其对α-葡萄糖苷酶的抑制效果尤为突出,TAPP浓度为2.0 mg/mL时,对α-葡萄糖苷酶的抑制率为(93.32±1.09)%,与阿卡波糖效果相近(P<0.05);TAPP给药浓度达到75μg/mL时,可以显著改善棕榈酸诱导的HepG2-IR模型细胞的胰岛素抵抗症状(P<0.05)。该研究结果为兰州地区金耳资源的充分开发和高值化利用提供了一定理论支持和开发基础。展开更多
为了探究金耳子实体胶质(Naematelia aurantialba fruit body glial,简称NAG)对小鼠血清细胞因子及肠道菌群的调节作用,分析NAG的成分和特征,并将其对正常小鼠进行为期21 d的灌胃处理,收集血清样本和粪便样本等,采用酶联免疫吸附法(ELI...为了探究金耳子实体胶质(Naematelia aurantialba fruit body glial,简称NAG)对小鼠血清细胞因子及肠道菌群的调节作用,分析NAG的成分和特征,并将其对正常小鼠进行为期21 d的灌胃处理,收集血清样本和粪便样本等,采用酶联免疫吸附法(ELISA)测定血清细胞因子含量,气-质联用(GC-MS)技术测定盲肠内容物短链脂肪酸(SCFAs)含量,16S rRNA测序技术分析小鼠粪便菌群并进行基因功能预测。结果显示:NAG主要营养成分为膳食纤维,其摄入使小鼠血清细胞因子TNF-α含量增加了1.21倍,IL-1β含量降低100%,IL-10含量增加了4.42倍;盲肠内容物中短链脂肪酸丁酸的含量显著增加(P<0.05);粪便中毛螺菌科(Lachnospiraceae)、瘤胃球菌科(Ruminococcaceae)相对丰度显著增加(P<0.05);肠道菌群基因功能中分泌系统、硫中继系统和谷氨酸能突触相对丰度显著增加(P<0.05),传染疾病、其他聚糖降解和脂多糖生物合成蛋白相对丰度显著降低(P<0.05)。表明NAG能够通过促进肠道益生菌群的增殖和短链脂肪酸丁酸的分泌,调节肠道菌群功能,使肠道微生态趋于健康,有助于提升免疫力、改善代谢和降低慢性病风险。展开更多
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.
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Extracellular polysaccharide from Tremella aurantialba (TEP) significantly inhibited H2O2-induced haemolysis of RBCs, significantly decreased the amount of malondialdehyde (MDA) produced in mice liver homogenates as a result of autoxidation, and inhibited VitC-Fe2+-induced swelling of liver mitochondria.
文摘以甘肃省兰州地区培育的金耳子实体为原料,经分离纯化得到分子质量(M_(w)=1.54×10^(6)Da)分布均一的金耳纯多糖(Tremella aurantialba pure polysaccharide,TAPP),其主要单糖组成为甘露糖∶葡萄糖醛酸∶葡萄糖∶木糖=55.20∶9.76∶14.81∶20.23,含有少量半乳糖和半乳糖醛酸。傅里叶变换红外光谱、刚果红实验、扫描电子显微镜分析和热重分析结果表明,TAPP含有β-糖苷键和螺旋结构,表面微观形貌呈片状无规则聚集状态,170℃以下热稳定性良好。活性分析结果显示,TAPP对α-淀粉酶的和α-葡萄糖苷酶的抑制与浓度呈正相关,其对α-葡萄糖苷酶的抑制效果尤为突出,TAPP浓度为2.0 mg/mL时,对α-葡萄糖苷酶的抑制率为(93.32±1.09)%,与阿卡波糖效果相近(P<0.05);TAPP给药浓度达到75μg/mL时,可以显著改善棕榈酸诱导的HepG2-IR模型细胞的胰岛素抵抗症状(P<0.05)。该研究结果为兰州地区金耳资源的充分开发和高值化利用提供了一定理论支持和开发基础。
文摘为了探究金耳子实体胶质(Naematelia aurantialba fruit body glial,简称NAG)对小鼠血清细胞因子及肠道菌群的调节作用,分析NAG的成分和特征,并将其对正常小鼠进行为期21 d的灌胃处理,收集血清样本和粪便样本等,采用酶联免疫吸附法(ELISA)测定血清细胞因子含量,气-质联用(GC-MS)技术测定盲肠内容物短链脂肪酸(SCFAs)含量,16S rRNA测序技术分析小鼠粪便菌群并进行基因功能预测。结果显示:NAG主要营养成分为膳食纤维,其摄入使小鼠血清细胞因子TNF-α含量增加了1.21倍,IL-1β含量降低100%,IL-10含量增加了4.42倍;盲肠内容物中短链脂肪酸丁酸的含量显著增加(P<0.05);粪便中毛螺菌科(Lachnospiraceae)、瘤胃球菌科(Ruminococcaceae)相对丰度显著增加(P<0.05);肠道菌群基因功能中分泌系统、硫中继系统和谷氨酸能突触相对丰度显著增加(P<0.05),传染疾病、其他聚糖降解和脂多糖生物合成蛋白相对丰度显著降低(P<0.05)。表明NAG能够通过促进肠道益生菌群的增殖和短链脂肪酸丁酸的分泌,调节肠道菌群功能,使肠道微生态趋于健康,有助于提升免疫力、改善代谢和降低慢性病风险。
