Use of Zebrafish Embryo Assay to Evaluate Toxicity and Safety of Bioreactor-Grown Exopolysaccharides and Endopolysaccharides from European Ganoderma applanatum Mycelium for Future Aquaculture Applications

Natural mycelial exopolysaccharide (EPS) and endopolysaccharide (ENS) extracted from bioreactor-cultivated European Ganoderma applanatum mushrooms are of potential high commercial value for both food and adjacent biopharmaceutical industries. In order to evaluate their potential toxicity for aquaculture application, both EPS (0.01–10 mg/mL) and ENS (0.01–10 mg/mL) extracts were tested for Zebrafish Embryo Toxicity (ZFET); early development effects on Zebrafish Embryos (ZE) were also analyzed between 24 and 120 h post-fertilization (HPF). Both EPS and ENS are considered non-toxic with LC₅₀ of 1.41 mg/mL and 0.87 mg/mL respectively. Both EPS and ENS did not delay hatching and teratogenic defect towards ZE with <1.0 mg/mL, respectively. No significant changes in the ZE heart rate were detected following treatment with the two compounds tested (EPS: 0.01–10 mg/mL: 176.44 ± 0.77 beats/min and ENS: 0.01–10 mg/mL: 148.44 ± 17.75 beats/min) compared to normal ZE (120–180 beats/min). These initial findings support future pre-clinical trials in adult fish models with view to safely using EPS and ENS as potential feed supplements for supplements for development of the aquaculture industry.     

灵芝菌丝体多糖的分离纯化及其单糖组成分析与分子量测定

前期杂交优化后赤芝菌种经液体深层发酵后,提取灵芝菌丝体多糖,并过DEAE-Sepharose Fast Flow柱分离纯化,利用高效体积排阻色谱(HPSEC)检测多糖级分的纯度,采用完全酸水解PMP柱前衍生化RP—HPLC测定多糖级分的单糖组成,多角度光散射仪联用装置(SEC—MALLS)测定其绝对重均分子量(Mw),并且根据分子旋转半径与分子摩尔数的关系曲线斜率初步推断其空间构象。结果显示:分离纯化得到3个多糖级分GLMP1、GLMP2和GLMP3,HPSEC检测其峰面积百分比分别为93.58%,97.64%,99.19%,单糖组成分析结果表明GLMP1、GLMP2和GLMP3均含有甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖和岩藻糖,但单糖摩尔比各异。SEC—MALLS测试GLMP1、GLMP2和GLMP3的Mw分别为4.526×105,4.603×104,3.760×103 g/mol,3个多糖级分构象可能均为高度紧缩且具有分支结构的聚合物。     

微波真空干燥高粘度的灵芝浓缩液

热敏性、高粘度物料的干燥仍是现代干燥技术中的一个难题。灵芝水提后，由薄膜蒸发器浓缩至含水量70％左右，然后采用微波真空干燥（真空度3000Pa）至水分含量10％左右，改用传统的电热真空干燥（55～60℃）至含水量6％～7％。对干燥产品的主要生物活性成分——灵芝多糖和三萜酸进行了分析检测并与其它干燥方法进行了比较。结果表明：利用微波真空干燥的灵芝产品其灵芝多糖和三萜酸的保留率与冷冻干燥产品十分接近。而比传统真空干燥（60—65℃）的产品要高得多，此外采用微波真空干燥的干燥时间要短得多。     

大孔树脂分离纯化女贞子中齐墩果酸工艺的研究

本文对大孔吸附树脂分离纯化女贞子中齐墩果酸的方法进行优化研究.以齐墩果酸的吸附率和解吸附率为考察指标,筛选出D101、D201、LSD001、XDA-7、C008、LSA-10、LSA-40、LSA-21等8种树脂中最适合齐墩果酸分离的树脂,并确定其分离纯化女贞子中齐墩果酸的最佳工艺条件.结果最优树脂为LSA-21型,最佳工艺条件为女贞子溶液浓度为4.0mg/mL,吸附流速为3mL/min,上样量2BV,洗脱剂为无水乙醇,解吸流速为3mL/min,洗脱剂用量为1.5BV.洗脱液经浓缩重结晶得结晶,经与齐墩果酸对照品进行TLC检查及熔点测定,确定其为齐墩果酸.     

18个灵芝菌种的酯酶同工酶分析

对18个灵芝菌株进行了菌丝培养观察、拮抗试验和酯酶同工酶聚丙烯酰胺凝胶电泳实验，并对它们作了聚类统计分析。分析结果表明：在相似水平为50％时供试菌株聚合为6类：第1类包括以韩芝为代表的7个菌株；第2类包括以赤芝为代表的5个菌株；第3类包括紫芝和黑芝；第4类为血芝和京大灵芝；第5类为白芝；第6类为日本赤芝。聚类分析结果与形态特征及拮抗试验结果基本一致。实验还说明酯酶同工酶可作为灵芝良种选育的一种有效标记方法，在菌种鉴定、保藏和管理方面具有重要作用。     

低档绿茶对灵芝菌摇瓶发酵的影响

研究了低档绿茶对灵芝菌摇瓶发酵的影响,低档绿茶由于成分多样复杂,一些特殊因子能够促进灵芝菌发酵过程中胞外多糖的合成,但因其多酚类等抗性物质的存在,一定程度上抑制了菌体生长,与对照实验相比,其生物量降低了17.08%,胞外粗多糖增加了42.5%.同时探讨了低档绿茶经过灵芝菌发酵作用后,茶汤中主要呈味成分的变化,实验结果表明:茶多酚降解了64.8%,游离氨基酸总量增加了9.7%、酚氨比降低了67.8%、水溶性灵芝胞外多糖增加,茶汤品质得到很大提高.     

Deproteinization and structural characterization of bioactive exopolysaccharides from Ganoderma sinense mycelium

Deproteinization was comparatively employed for the purification of crude exopolysaccharide (CEP) of Ganoderma sinense mycelium. Results showed that ammonium sulfate (70% saturation) salting out could remove 95% proteins of CEP, whereas 20% trichloroacetic acid precipitation was best with a polysaccharide loss less than 20%. Protein-bound polysaccharides could be removed by both methods, and polysaccharides with high molecular weight dominated in the lost polysaccharides. Through anion-exchange chromatography, a highly branched α-(1→6)-D-mannan and a protein-bound heteropolysaccharide were prepared from CEP. Both fractions could inhibit the in vitro proliferation of tumor cells BEL-7402, and induce TNF-α secretion of murine splenocytes.     

Maximizing the Retention of Ganoderic Acids and Water-Soluble Polysaccharides Content of Ganoderma lucidum Using Two-Stage Dehydration Method

Ganoderic acids and water-soluble polysaccharides are the main bioactive ingredients of Ganoderma lucidum and are heat sensitive. Hence, it is crucial to apply a suitable drying method for the drying of G. lucidum to minimize the loss of these bioactive ingredients. In this study, a two-stage drying method was applied to enhance the drying kinetics, as well as the retention of both ganoderic acids and water-soluble polysaccharides in dried G. lucidum fruiting bodies and slices compared to other single-stage-dried products. The results showed that two-stage drying enhanced the overall drying rate, which consequently shortened the total drying time up to 61 and 330% for fruiting bodies and slices, respectively, compared to single-stage drying methods. Furthermore, two-stage drying of Ganoderma fruiting bodies, which involves 18-h vacuum drying followed by 5-h heat pump drying, maximized the retention of ganoderic acids and water-soluble polysaccharides, with the retention of 97.90% of water-soluble polysaccharides and 4.2% more ganoderic acids than freeze-dried products. On the other hand, drying of Ganoderma slices within the shortest total drying time, which is 10-min heat pump drying followed by 50-min oven drying, also retained most of the ganoderic acids and water-soluble polysaccharides in the dried products; that is, about 80% of water-soluble polysaccharides and 11% more ganoderic acids than freeze-dried slices.     

灵芝孢子油对四氯化碳联合高脂饮食诱导大鼠肝纤维化的作用研究

探究灵芝孢子油(GSO)对四氯化碳(CCl₄)联合高脂饮食诱导的大鼠肝纤维化的改善作用及机制。将雄性SPF级SD大鼠分为空白对照组、模型对照组、阳性药物组和灵芝孢子油低、中、高剂量组(人体推荐量的5、15、30倍)。采用CCl₄联合高脂饲料诱导6周建立大鼠肝纤维化模型,造模过程中灌胃GSO干预8周。通过检测血清生理生化指标和大鼠肝脏组织形态、氧化应激水平、纤维化相关蛋白,评价GSO对肝纤维化的保护作用。研究结果表明,中、高剂量GSO能够显著提升肝纤维化大鼠的存活率,使得大鼠存活率恢复至空白对照组水平。病理染色结果表明,GSO处理的大鼠肝组织纤维化得到不同程度的改善,GSO中、高剂量处理显著降低大鼠肝脏胶原面积占比及面密度。GSO高剂量干预可显著降低大鼠血浆肝功能相关指标(谷丙转氨酶、谷草转氨酶、总胆红素、碱性磷酸酶)水平,减少血浆中肝纤维化指标(透明质酸、层黏蛋白、Ⅳ型胶原、羟脯氨酸)水平。同时,高剂量GSO干预还显著降低肝组织中丙二醛含量,提高还原型谷胱甘肽含量与超氧化物歧化酶活性,显著升高基质金属蛋白酶-9的表达。研究结果证实,中、高剂量...     

我国灵芝精深加工研究进展

灵芝是我国传统中医中药的瑰宝,具有极高的药用价值,有抗肿瘤、增强免疫功能、降血糖、保肝、抗衰老、抗炎、抗凝血等作用。灵芝产业发展迅速,产品形式多样,有药品、保健食品、化妆品和其他生物敷料等,全球范围内灵芝产品年产值已超过25亿美元。灵芝从中国古代神话进入普通人的生活,从中华大地走向世界各地,在保护人类健康,造福人类的同时,也创造了巨大财富。本文就灵芝精深加工现状、存在问题及发展趋势进行综述,为灵芝产业持续稳定快速发展提供基础。