Interaction of hispidin with pepsin:Multi-spectroscopic analyses and docking simulation

Hispidin is a pyranone compound found in edible and medicinal mushrooms of the Phellinus and Inonotus genera.This investigation used fluorescence spectroscopy,UV absorption spectroscopy,and molecular docking to examine the interaction of hispidin with pepsin.The Stern-Volmer method was used to perform the fluorescence quenching measurements at different temperatures(298 K,303 K,and 310 K).According to the findings,hispidin induced a static quenching mechanism in pepsin that resulted in the creation of a hispidin-pepsin complex with binding constants(Ka)ranging from 9.56×10^(4) to 3.45×10^(5) L mol^(-1).The positive values ofΔH(84.6 kJ mol-1)andΔS(337.9 J mol^(-1) K^(-1))demonstrated that hydrophobic forces contributed to forming the hispidin-pepsin complex.The findings of UV-vis absorption,synchronous fluorescence,and 3D fluorescence spectraspectra demonstrated that hispidin altered the conformation and microenvironment of pepsin.According to the analysis of molecular docking,hispidin got into the pepsin's active cavity.The research clarifies the molecular mechanisms by which hispidin binds to pepsin and helps understand its possible biological activity in vivo.

Production of hispidin polyphenols from medicinal mushroom Sanghuangporus vaninii in submerged cultures

Objective: The medicinal mushroom Sanghuangporus vaninii produces pharmaceutically valuable hispidin polyphenols in natural habitats. However, due to the slow growth in nature, S. vaninii grown in the field(sclerotia) is not reliable for pharmaceutical purposes. Although higher biomass of fungal mycelia can be obtained in submerged cultures, the accumulation of hispidin polyphenols is rare.Methods: In this study, the polyunsaturated fatty acids(PUFAs), linoleic acid(LA), linolenic acid(ALA),and methyl jasmonate(Me Ja) were employed as the stimulant agents to coordinate the accumulation of biomass and hispidin polyphenols in its submerged cultures.Results: The addition of LA and ALA promoted the mycelial accumulation, while the addition of Me Ja inhibited the growth of S. vaninii concomitant with reduced total polyphenols. UPLC-Triple-TOF-MS analysis revealed an increased production of hispidin, phellinstatin, pinnilidine, and its derivatives upon the addition of LA and ALA, and hypholomine B and its isomer, 3,14’-bihispidinyl, and phelligridin E upon the addition of Me Ja on day 13. Intriguingly, total polyphenols from the Me Ja-supplementing cultures harbored a high capacity in scavenging free radicals. Chemical structural analysis showed that hispidin polyphenols had higher antioxidant activity due to more hispidin moieties induced by Me Ja.Conclusion: The supplement of PUFAs affects the synthesis and composition of hispidin polyphenols in S.vaninii. Our results provide a possibility to coordinate the production of hispidin polyphenols via submerged cultures of S. vaninii.

裂蹄木层孔菌菌丝体培养物中的β-分泌酶抑制剂hispidin

阿尔茨海默病(AD)的主要特征是淀粉样蛋白沉积(淀粉样斑块)，淀粉斑块主要成分是β-淀粉肽(Ap)。Aβ的形成与β-分泌酶有直接关系，而α-分泌酶则可防止Aβ的形成。因此，抑制β-分泌酶和诱导α-分泌酶可能是减少脑中Aβ形成的标靶。作者测试了256种植物和真菌提取物，从裂蹄木层孔菌Phellinus linteus Teng菌丝体培养液中分离、鉴定了一种小分子β-分泌酶(BACEl)抑制剂hispidin，

茉莉酸和双氧水对斜生褐孔菌一株黑色素缺失突变体产生酚类化合物的影响(英文)

黑色素合成是桦褐孔菌Phaeoporus obliquus次生代谢过程中的一个重要事件。黑色素的过多合成势必影响其他酚类的积累。本文以茉莉酸和双氧水为调控因子,开展了该突变体深层发酵过程酚类化合物积累研究。正常培养条件下,每升发酵液积累酚类化合物最高可达317.45mg/L,其中包括苯甲酸衍生物(45.3mg/L),黄酮苷元(74.4mg/L),黄酮苷(160mg/L)以及硬毛素类化合物(18.5mg/L)。加入茉莉酸后,总酚最大值增加到620.45mg/L,其中苯甲酸衍生物增加到230.26mg/L,黄酮苷元增加到166.86mg/L,而黄酮苷和硬毛素类化合物却分别降低到98.5mg/L和7.1mg,双氧水的加入没有提高总酚的最大积累(315.69mg/L)。因此,与野生型相比,黑色素缺失使总酚的积累有所增加,随茉莉L酸的加入总酚积累最大值增加近一倍。虽然双氧水的加入提高了苯丙氨酸裂解酶活性,但总酚的积累却没有增加,这可能是由于一部分酚类化合物在抗双氧水氧化胁迫过程中被消耗。

深层培养的斜生褐孔菌酚类化合物对铅处理小鼠耐力的增强作用(英文)

本文研究了不同条件下深层培养的斜生褐孔菌Phaeoporus obliquus与野生桦褐孔菌在酚类化合物组成及其含量上的差异以及对铅处理小鼠常压缺氧和游泳耐力影响。野生桦褐孔菌主要含有水溶性黑色素、硬毛素衍生物,两者各占总酚类化合物的12.1%和42.2%。而深层培养的桦褐孔菌则主要积累黄酮类化合物,其中正常生理条件下积累量达到总酚的35.5%,加入H2O2后积累量达到总酚的92.5%,同时加入H2O2和熊果苷后则为总酚的76.9%。正常培养基中水溶性黑色素的积累量达到总酚的20.2%,而加入H2O2或同时加入H2O2和熊果苷后水溶性黑色素则很少合成。口服野生斜生褐孔菌或不同条件下深层培养的斜生褐孔菌酚类化合物都能使铅处理小鼠忍耐缺氧和游泳的时间延长12–15min。同时,口服不同来源的斜生褐孔菌酚类化合物可提升铅处理小鼠外周淋巴细胞的活力和总超氧化物歧化酶活性,并降低丙二醛的积累。因此,斜生褐孔菌酚类化合物对铅处理小鼠耐缺氧和游泳的提升作用在于抑制了脂质过氧化反应,增强了过氧化物歧化酶的表达,从而保护了铅处理小鼠的细胞和器官免于氧化胁迫的损伤。

液态发酵桑黄菌丝体抗A型流感(H1N1)病毒之效用评估

本研究旨在利用细胞试验评估液态发酵桑黄(Sanghuangporus sanghuang)菌丝体萃取物之抗A型流感病毒(H1N1)的能力,并进而分析活性所在之分层。实验使用MTS法评估桑黄菌丝体萃取物分别在预防、共培养以及治疗三种试验中减轻病毒对于宿主细胞的毒杀性的效果。结果显示酒萃物优于水萃物,再将酒萃物进行层析分离,共获得四种分层,分别为水层(PlH2O)、1-丁醇层(PlBtOH)、二氯甲烷层(PlDCM)以及己烷层(PlHex)。其中,以PlBtOH、PlDCM两组在预防试验、共培养试验以及治疗试验中,皆可见显著提升细胞存活率之效果(p 【0.05)。对比四分层的UPLC图谱后发现,PlBtOH及PlDCM内皆含有较高的hispidin,PlBtOH另含有hypholomine B (HB),抗病毒试验中则以PlBtOH最强,PlDCM次之,因此推估液态发酵桑黄菌丝体之抗H1N1流感病毒功效可能来自于hispidin以及HB。

中药桑黄抗肿瘤作用及其机制研究进展

桑黄是一类珍贵的药用真菌,近年来,随着现代生命科学、分子生物学等技术的发展,桑黄的药用有效成分和药用机制被进一步发现、研究和应用,尤其发现其抗肿瘤作用明显。本文综述了桑黄的各种成分提取物及其抗肿瘤作用机制,为桑黄的开发利用及进一步相关研究提供思路与参考。

桦褐孔菌酚类化合物及其免疫促进作用(英文)

Phenolic compounds from field-grown Inonotus obliquus sclerotia (Chaga) consist mainly of hispidin analogs and melanins, and are thought to be the active constituents to treat several human diseases. In submerged cultures of the fungus, however, no information is currently available on the production of phenolic compounds and their corresponding pharmacological functions. In this study, phenolic compounds from Chaga and submerged cultures of the fungus were assayed for their composition and immune-stimulating effects. Phenolic compounds produced by I. obliquus in submerged cultures mostly consist of flavonoids, together with small amounts of hispidin analogs and melanins. This is quite contrary to the situation in Chaga, where flavonoids are determined as trace elements. Furthermore, phenolic compounds from Chaga show capacity about two-fold higher than those produced in submerged cultures in inhibiting cyclophosphamide-induced reduction of bodyweight, spleen index and viability of peripheral lymphocytes in test mice. Thus less production of hispidin analogs and melanins is likely to be responsible for less immune-stimulating effects in phenolic compounds from submerged cultures, and additional factors should be imposed during submerged cultures of I. obliquus to regulate biosynthesis of phenolic compounds directed to the composition similar to Chaga.

一氧化氮对液体发酵瓦尼桑黄多酚和三萜积累的影响

本文旨在研究外源一氧化氮对瓦尼桑黄多酚和三萜类化合物积累的影响。以发酵罐发酵法培养瓦尼桑黄,并在培养液中添加硝普钠。采用Folin-Ciocalteu法测定瓦尼桑黄菌丝体内多酚类化合物的含量并以高效液相色谱法(HPLC)测定多酚的组成成分。利用香草醛-冰醋酸-高氯酸分光光度计法测定三萜类化合物的含量。结果显示,硝普钠的添加可以促进瓦尼桑黄菌丝体的生长,提高菌丝体内次级代谢产物清除1,1-二苯基-2-三硝基苯肼(DPPH)自由基的能力。硝普钠的添加使瓦尼桑黄中总多酚和三萜类化合物的积累分别增长了14.3%和51.7%。HPLC分析发现,发酵培养3–7d,硝普钠添加组培养物中硬毛素、纤孔菌素A和桑黄素D含量均增加。可见,一氧化氮对液体发酵条件下瓦尼桑黄多酚和三萜类化合物的合成具有明显的调节作用。

鲍氏层孔菌子实体的化学成分研究

采用硅胶和Sephadex LH20柱层析方法,从鲍氏层孔菌子实体提取物中分离得到8个化合物。运用NMR和MS等波谱法分析和鉴定为7(8),22(23)-二烯-3-酮-麦角甾烷、4,6,8(14),22(23)-四烯-3-酮-麦角甾烷、麦角甾醇、过氧化麦角甾醇、三十烷酸对羟基苯乙酯、4-(3,4-二羟苯基)-3-丁烯-2-酮、hispolon、hispidin。