CN114957496B - 一种冠突曲霉菌孢子多糖及其制备方法和应用 - Google Patents
一种冠突曲霉菌孢子多糖及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种冠突曲霉菌孢子多糖及其制备方法和应用,制备方法包括:将冠突曲霉菌孢子破壁后进行干燥,研磨成粉末,按料液比为1g∶20‑50mL的比例与低共熔溶剂混匀,在70℃条件下反应30‑55min,随后抽滤取滤液;将滤液加入到乙醇溶液中,静置醇沉后离心,收集沉淀物;将沉淀物用蒸馏水溶解,再用Sevag法脱蛋白,所得溶液经过透析后真空冻干,即得冠突曲霉菌孢子多糖。试验结果表明,本发明所提供的冠突曲霉菌孢子多糖具有良好的改善代谢综合征的效果,包括:改善代谢综合征导致的体重增加、脂肪积累、肝脏脂质积累和肝功能障碍、胰岛素抵抗、血脂紊乱、机体炎症,保护结肠屏障功能,恢复由代谢综合征导致的肠道菌群失衡状态,调节肠道菌群。
Description
技术领域
本发明属于微生物发酵、食品及医药技术领域,更具体地,涉及一种冠突曲霉菌孢子多糖及其制备方法和应用。
背景技术
代谢综合征(Metabolic syndrome,MS)是一组以脂质代谢紊乱、血脂异常、肝脏脂质堆积和胰岛素抵抗为特征的代谢紊乱疾病,同时会增加患肥胖症、高血脂症、非酒精性脂肪肝、2型糖尿病、癌症和心血管疾病的风险。
近年来,随着人们生活水平的提高、饮食结构中油脂和碳水化合物比例的增加、缺乏锻炼、吸烟以及过量饮酒等不良行为都导致了国内代谢综合征患病率的显著增加,俨然已经成为了严重的公共卫生问题。因此,寻求一种合适的能够缓解代谢综合征的手段是十分必要的。
目前,针对代谢综合征的治疗手段包括加强运动、改善饮食和药物治疗;所服用的药物包括减肥药物奥利司他、双胍类降糖药物、降血脂药物辛伐他丁片等等,这些药物对代谢综合征单一的指标具有良好的效果,但是,长期服用会出现一定的副作用。具体而言,奥利司他有腹泻的副作用,还对肾脏有负面影响,加之在全球4000万奥利司他使用者中,已报告13例严重肝损伤;双胍类降糖药物副作用有乏力、恶心、呕吐及腹泻,可因乳酸过多引起乳酸中毒。因此人们渴望一种安全、轻松、有效地缓解代谢综合征手段。
据研究报道,机体内肠道微生物的紊乱与代谢综合征有关,并且已经证实高脂饮食破坏肠道屏障功能和肠道菌群紊乱,增加机体内循环的内毒素水平,诱发全身性炎症,最终导致代谢紊乱。因此,肠道菌群被认为是代谢综合征治疗的新靶点。
近些年来,真菌多糖(如冬虫夏草多糖、灵芝多糖和茯苓多糖)已被证明能够通过调节肠道菌群来改善代谢综合征。冠突曲霉菌(Aspergillus cristatus)是茯砖茶发花过程中唯一优势真菌,最终在茯砖茶内部生成大量金黄色颗粒,俗称“金花”。由于冠突曲霉菌子囊孢子有一层较厚的孢子壁,其质地坚韧、不易分解,阻碍了人们对其中物质的有效利用。目前对冠突曲霉菌孢子多糖在改善代谢综合征以及如何保护肠道屏障、调节肠道菌群方面少有研究,因此具有潜在地研究开发价值。
鉴于此,特提出本发明。
发明内容
针对现有技术所存在的不足,本发明提供了一种冠突曲霉菌孢子多糖及其制备方法和应用。本发明为了评估冠突曲霉菌孢子多糖改善代谢综合征的作用,通过对代谢综合征模型的动物的干预作用来评价冠突曲霉菌孢子多糖改善代谢综合征的效果。
为实现上述目的,本发明的技术方案如下:
一种冠突曲霉菌孢子多糖的制备方法,包括以下步骤:
S1、将冠突曲霉菌孢子破壁后进行干燥,研磨成粉末,按料液比为1g:20-50mL的比例与低共熔溶剂混匀,在70℃条件下反应30-55min,随后抽滤取滤液;
S2、将滤液加入到乙醇溶液中,静置醇沉后离心,收集沉淀物;
S3、将沉淀物用蒸馏水溶解,再用Sevag法脱蛋白,最后将所得溶液经过透析后真空冻干,即得冠突曲霉菌孢子多糖;
其中,所述低共熔溶剂为氯化胆碱与草酸按摩尔比1∶1混合后加热制得。
在上述技术方案中,步骤S2中,所述乙醇溶液为85-98v%的乙醇水溶液。
在本发明的一个优选实施方式中,步骤S2中,所述乙醇水溶液的加入体积为滤液的3-5倍。
进一步地,在上述技术方案中,步骤S2中,所述静置醇提具体为,置于2-6℃的冰箱中静置24-48h。
进一步地,在上述技术方案中,步骤S2中,所述离心的转速为5000-8000rpm。
在上述技术方案中,步骤S1中,所述反应的时间为40min。
本发明另一方面还提供了上述制备方法制备得到的冠突曲霉菌孢子多糖。
具体地,在上述技术方案中,上述冠突曲霉菌孢子多糖的单糖组成为核糖、葡萄糖、半乳糖和甘露糖,且核糖、葡萄糖、半乳糖和甘露糖的摩尔比为1∶1.7∶4.4∶5.2。
本发明又一方面还提供了上述冠突曲霉菌孢子多糖在制备改善代谢综合征的药物和功能性食品中的应用。
具体地,在上述技术方案中,在所述改善代谢综合征的药物和功能性食品中,冠突曲霉菌孢子多糖的含量为0.1-99.5wt%。
具体地,在上述技术方案中,所述冠突曲霉菌孢子多糖具有良好的改善代谢综合征的效果。
本发明与现有技术相比,具有以下优点:
(1)本发明所提供的冠突曲霉菌孢子多糖的制备方法中,所采用的提取溶剂为低共熔溶剂,其是一种绿色溶剂,具有原材料充足易得、价格低廉、毒性较低和污染少等优点;
(2)本发明所提供的冠突曲霉菌孢子多糖具有如下生物学特性:
能够显著改善代谢综合征导致的体重增加,能够显著改善代谢综合征导致的脂肪积累,能够显著改善代谢综合征导致的肝脏脂质积累和肝功能障碍,能够显著改善代谢综合征大鼠的胰岛素抵抗,能够显著改善代谢综合征大鼠的血脂紊乱,能够显著改善代谢综合征大鼠的机体炎症,对保护结肠屏障功能具有良好的效果,可显著恢复由代谢综合征导致的肠道菌群失衡状态;
(3)本发明所提供的冠突曲霉菌孢子多糖的改善代谢综合征的效果通过粪便移植试验进一步证实了冠突曲霉菌孢子多糖通过调节肠道菌群来改善代谢综合征。
附图说明
图1为本发明实施例中冠突曲霉菌孢子多糖对代谢综合征大鼠的体重增加量、肾周脂肪和附睾脂肪重量的影响以及附睾脂肪H&E染色;
图2为本发明实施例中冠突曲霉菌孢子多糖对代谢综合征大鼠的肝脏重量/体重、谷草转氨酶(AST)、谷丙转氨酶(ALT)的影响以及肝脏H&E染色和油红O染色;
图3为本发明实施例中冠突曲霉菌孢子多糖对代谢综合征大鼠的胰岛素抵抗指数(HOMA-IR)的影响;
图4为本发明实施例中冠突曲霉菌孢子多糖对代谢综合征大鼠的血清水平上总甘油三酯(TC)、总胆固醇(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)的影响;
图5为本发明实施例中冠突曲霉菌孢子多糖对代谢综合征大鼠的血清水平上人巨噬细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子α(TNF-α)、白介素6(IL-6)、白介素-10(IL-10)和脂多糖(LPS)的影响;
图6为本发明实施例中冠突曲霉菌孢子多糖对代谢综合征大鼠的结肠组织H&E染色和PAS染色;
图7为本发明实施例中冠突曲霉菌孢子多糖对代谢综合征大鼠肠道菌群的影响;
图8为本发明实施例中粪便移植对代谢综合大鼠的影响;
图9为本发明实施例中粪便移植对代谢综合征大鼠肠道菌群的影响;
注:不同的小写字母表示差异显著(p<0.05)。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合实施例,对本发明进行进一步详细说明。
应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。
实施例中,如无特别说明,所用手段均为本领域常规的手段。
本文中所用的术语“包含”、“包括”或其任何其它变形,意在覆盖非排它性的包括。例如,包含所列要素的组合物、步骤、方法、制品或装置不必仅限于那些要素,而是可以包括未明确列出的其它要素或此种组合物、步骤、方法、制品或装置所固有的要素。
实施例中未注明具体技术或条件者,按照本领域内的文献所描述的技术或条件,或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可通过正规渠道商购买得到的常规产品。
在本文中所披露的范围的端点和任何值都不限于该精确的范围或值,这些范围或值应当理解为包含接近这些范围或值的值。对于数值范围来说,各个范围的端点值之间、各个范围的端点值和单独的点值之间,以及单独的点值之间可以彼此组合而得到一个或多个新的数值范围,这些数值范围应被视为在本文中具体公开。
此外,下面所描述的本发明各个实施方式中所涉及到的技术特征只要彼此之间未构成冲突就可以相互组合。
在本发明实施例中,所用的冠突曲霉菌孢子是通过将冠突曲霉菌Aspergilluscristatum GTQM2021-01(于2021年10月26日保藏于中国典型培养物保藏中心,保藏地址:中国.武汉.武汉大学,邮编430072,保藏编号为CCTCC NO:M20211325)接种于黑毛茶后进行散茶发花,然后将生长在茶叶中的冠突曲霉菌孢子(即闭囊壳)经过振摇后,过筛去除杂质所得。
实施例1
本发明实施例提供了冠突曲霉菌孢子多糖的提取、分离和纯化方法,包括如下步骤:
(1)将冠突曲霉菌孢子采用超声破壁,冷冻干燥后研磨成粉末,按料液比为1g:20-50mL的比例与低共熔溶剂(氯化胆碱与草酸按摩尔比1:1加热混合制成)混匀,在70℃条件下反应40min,然后抽滤后取滤液;
(2)取步骤(1)中的滤液,加入3-5倍的95v%的乙醇水溶液,在4℃冰箱内放置24-48h醇沉,采用4000-8000r/min离心15分钟收集沉淀物;
(3)取步骤(2)中的沉淀,用蒸馏水溶解,再用Sevag法脱蛋白,所得溶液经过透析后,在真空冷冻干燥机中冻干,得到冠突曲霉菌孢子多糖。
经检测,冠突曲霉菌孢子多糖的单糖组成为:核糖∶葡萄糖∶半乳糖∶甘露糖=1∶1.7∶4.4∶5.2。
试验验证
所用的实验动物的饲养方法为:
1、选用雄性SPF级4周龄健康的雄性Sprague Dawley(SD)大鼠,体重200±10g;
2、实验大鼠喂养在湖南农业大学标准动物房(SPF级),在24-25℃环境中,开灯时间和关灯时间为12h/12h循环,每4只大鼠置于一个鼠笼中,鼠笼中有辐照杨木垫料覆盖,整个实验过程中自由进食与饮水,饲料每天更换一次并且记录投放量与剩余量,蒸馏水每天更换一次,笼中的垫料每隔48h更换一次以保持笼内整洁卫生;
3、正常饲料(Normal Diet,ND)和高脂饲料(High-fat Diet,HFD)均是购买于江苏南通特洛菲饲料有限公司(南通市,江苏省,中国)。其中:正常饲料(编号为TP23302)每克提供3.6kcal/g,由蛋白质19.4%,碳水化合物70.6%,脂肪10%;高脂饲料(编号为TP23300)每克提供5.1kcal/g,含蛋白质19.4%,碳水化合物20.6%,脂肪60%;
实验过程:
取体重为200±10g的雄性SPF级4周龄健康的雄性SD大鼠24只,适应性喂养1周,随机分为正常对照组(ND组)、代谢综合征组(HFD组)和冠突曲霉菌孢子多糖组(ACP组),每组有大鼠8只;其中:ND组大鼠饲喂正常饲料,灌胃蒸馏水;HFD组大鼠饲喂高脂饲料,灌胃蒸馏水;ACP组大鼠饲喂高脂饲料,灌胃浓度300mg/kg的冠突曲霉菌孢子多糖水溶液;灌胃时间为每天上午,灌胃体积为2mL,整个动物实验周期为8周(不包括适应性喂养)。
在0-8周内每周给所有大鼠称一次体重,同时,每天记录食物的消耗量。在灌胃试验结束时,用戊巴比妥将大鼠安乐死,称量肝脏组织重量、附睾脂肪和肾周脂肪组织重量,收集肝脏组织、结肠组织、附睾组织和血清样品进行后续分析;此外,在灌胃处理的最后一天在无菌环境下收集大鼠粪便,用液氮速冻之后,放入-80℃冰箱保存。
1、冠突曲霉菌孢子多糖对代谢综合征大鼠的体重过度增长抑制作用
实验结果如图1A所示。代谢综合征组(HFD组)大鼠体重增长量显著高于正常对照组(ND组),通过冠突曲霉菌孢子多糖的干预之后,大鼠体重增长量显著降低,且显著低于正常对照组(ND组)。上述结果说明,冠突曲霉菌孢子多糖对代谢综合征大鼠的体重过度增长显著抑制作用。
2、冠突曲霉菌孢子多糖对代谢综合征大鼠的脂肪积累抑制作用
实验结果如图1B-1D所示。冠突曲霉菌孢子多糖组和正常对照组大鼠的肾周脂肪(图1B)和附睾脂肪(图1C)重量显著低于代谢综合征组大鼠,且冠突曲霉菌孢子多糖组和正常对照组两者之间没有显著差异;附睾脂肪苏木精和曙红(H&E)染色切片结果(图1D)显示,冠突曲霉菌孢子多糖组的脂肪细胞体积显著小于代谢综合征组,脂肪细胞数目显著多于代谢综合征组。以上结果说明,冠突曲霉菌孢子多糖能够抑制代谢综合征大鼠的脂肪积累。
3、冠突曲霉菌孢子多糖改善代谢综合征导致的肝脏脂质积累和肝功能障碍
实验结果如图2所示。代谢综合征组大鼠的肝脏重量/体重(图2A)显著高于正常对照组,灌胃冠突曲霉菌孢子多糖后,肝脏重量/体重显著降低;同样,冠突曲霉菌孢子多糖组的谷草转氨酶(AST,图2B)和谷丙转氨酶(ALT,图2C)含量显著低于代谢综合征组;根据油红O染色(图2D)和H&E染色(图2E)切片可以得知,代谢综合征组大鼠的肝脏脂质积累严重,出现大小不一的脂滴空泡,肝小叶炎症严重,经过灌胃冠突曲霉菌孢子多糖后,肝脏脂滴空泡显著减少,炎症程度减轻,肝细胞结构完整。以上结果表明,冠突曲霉菌孢子多糖改善代谢综合征导致的肝脏脂质积累和肝功能障碍。
4、冠突曲霉菌孢子多糖缓解代谢综合征大鼠的胰岛素抵抗
大鼠安乐死时,用罗氏血糖仪测得大鼠空腹血糖以及通过试剂盒测得空腹胰岛素,根据胰岛素抵抗指标计算公式:HOMA-IR=(空腹胰岛素×空腹血糖值)/22.5。
实验结果如图3所示。代谢综合征组大鼠的HOMA-IR值显著高于正常对照组大鼠,冠突曲霉菌孢子多糖干预后,HOMA-IR值显著降低。结果表明,冠突曲霉菌孢子多糖能够缓解代谢综合征大鼠的胰岛素抵抗。
5、冠突曲霉菌孢子多糖改善代谢综合征大鼠的血脂紊乱
采取主动脉取全血样品,并且在凝固30分钟后,在4℃用离心机以2,500×g离心10分钟以获得血清。按照试剂盒的检测方法测定总甘油三酯(TC)、总胆固醇(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)的含量。
实验结果如图4所示。代谢综合征组的大鼠TC(图4A)、TG(图4B)和LDL-C(图4C)含量显著高于正常对照组,HDL-C(图4D)含量显著低于正常对照组;经过冠突曲霉菌孢子多糖干预之后,TC、TG和LDL-C含量显著降低,HDL-C含量显著升高。说明冠突曲霉菌孢子多糖能够缓解代谢综合征大鼠的血脂紊乱。
6、冠突曲霉菌孢子多糖改善代谢综合征大鼠的机体炎症
采取主动脉取全血样品,并且在凝固30分钟后,在4℃用离心机以2,500×g离心10分钟以获得血清。按照试剂盒的检测方法测定人巨噬细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子α(TNF-α)、白介素6(IL-6)、白介素-10(IL-10)和脂多糖(LPS)的含量。
实验结果如图5所示。代谢综合征大鼠血清中的MCP-1(图5A),TNF-α(图5B),IL-6(图5C)和LPS(图5E)的含量显著高于正常对照组,IL-10(图5D)含量显著低于正常对照组;经过冠突曲霉菌孢子多糖干预之后,MCP-1、TNF-α、IL-6和LPS含量显著降低,IL-10含量显著升高。说明冠突曲霉菌孢子多糖能够改善代谢综合征大鼠的机体炎症。
7、冠突曲霉菌孢子多糖恢复代谢综合征大鼠的肠道屏障损伤
将结肠组织在4%多聚甲醛中固定24小时,脱水后用石蜡包埋,然后将石蜡包埋的结肠组织切片后用苏木精和曙红(H&E)和过碘酸雪夫(Periodic Acid-Schiff,PAS)染色,脱水封片之后用光学显微镜采集图片分析。
实验结果如图6所示。结肠的H&E染色图(图6A)显示,代谢综合征组大鼠结肠表现出上表皮被破坏,水肿,肠绒毛的断裂,以及黏膜和黏膜下大面积炎症浸润。冠突曲霉菌孢子多糖组则表现出轻微的炎症反应,杯状细胞基本恢复正常,水肿得到明显改善,并且没有炎症细胞浸润。结肠的PAS染色图(图6B)显示,与正常对照组相比,代谢综合征大鼠结肠的PAS阳性区域面积显著降低,这说明代谢综合征大鼠结肠中产生黏蛋白的杯状细胞数目少,肠道屏障严重受损。灌胃冠突曲霉菌孢子多糖之后,PAS阳性区域显著增加。上述结果说明,冠突曲霉菌孢子多糖恢复代谢综合征大鼠的肠道屏障损伤。
8、冠突曲霉菌孢子多糖对代谢综合征大鼠的肠道菌群失衡具有恢复作用
对实验结束后的各种大鼠的粪便样品,采用16S rRNA测序技术及相应的生物信息学分析手段,来探究冠突曲霉菌孢子多糖对代谢综合征大鼠肠道菌群的生物多样性、菌群组成和结构的影响。
实验结果如图7所示。冠突曲霉菌孢子多糖能够恢复高脂饮食导致的肠道菌群结构失调,并且能够降低Firmicutes的相对丰度,提高Bacteroidetes的相对丰度以及Firmicutes/Bacteroidetes的比值。在属水平上,冠突曲霉菌孢子多糖能够显著提高代谢综合征大鼠体内Akkermansia,Bacteroides,Clostridium_sensu_stricto_1,Faecalibaculum,Parabacteroides,Ruminococcaceae_UCG-005和Rombousita等有益菌的相对丰度,抑制Blautia,Desulfovibrio,Lachnoclostridium,Roseburia和Streptococcus等有害菌的相对丰度。冠突曲霉菌孢子多糖灌胃的代谢综合征大鼠体内富集Akkermansia,Faecalibaculum和Lactobacillus等有益菌。以上结果说明,冠突曲霉菌孢子多糖能够调节代谢综合征大鼠体内的肠道菌群朝着健康的方向发展。
9、粪便移植试验进一步证明冠突曲霉菌孢子多糖通过调节肠道菌群来改善代谢综合征
为了进一步验证冠突曲霉菌孢子多糖改善代谢综合征和调节肠道菌群的因果关系,我们进行了粪便移植的实验。
具体实验方案如下:
每天收集正常对照组(ND组)、代谢综合征组(HFD组)和冠突曲霉菌孢子多糖组(ACP组)大鼠的新鲜粪便用于粪便移植。
将24只8周龄SD雄性大鼠随机分为三组:ND→HFD组(接受ND组大鼠的粪菌)、HFD→HFD组(接受HFD组大鼠的粪菌)和ACP→HFD组(接受ACP组大鼠的粪菌),每一组8只大鼠。将大鼠喂养在湖南农业大学标准动物房(SPF级),在24-25℃环境中,开灯时间和关灯时间为12h/12h循环,每4只大鼠置于一个鼠笼中,鼠笼中有辐照杨木垫料覆盖,整个实验过程中自由进食与饮水,饲料每天更换一次并且记录投放量与剩余量,蒸馏水每天更换一次,笼中的垫料每隔48h更换一次以保持笼内整洁卫生;三组大鼠均饲喂高脂饲料;实验周期为8周。
在0-8周内每周给所有大鼠称一次体重以及每天记录食物的消耗量。在灌胃试验结束时,用戊巴比妥将大鼠安乐死,称量肝脏组织重量、附睾脂肪和肾周脂肪组织重量;收集肝脏组织、结肠组织、附睾组织和血清样品进行后续分析。此外,在灌胃处理的最后一天在无菌环境下收集大鼠粪便,用液氮速冻之后,放入-80℃冰箱保存。
检测空腹血糖值、血清中AST、ALT、TC、TG、LDL-C、HDL-C、MCP-1、TNF-α、IL-6、IL-10、LPS和胰岛素含量;对附睾脂肪组织、肝脏组织以及结肠组织进行H&E染色,对肝脏组织进行油红O染色和对结肠组织进行PAS染色(如图8所示);采用16S rRNA测序技术及相应的生物信息学分析手段检测粪便移植对代谢综合征大鼠的肠道菌群影响(如图9所示)。
从图8中可以看出,与移植来自于代谢综合征大鼠的粪菌组来比,移植来自于冠突曲霉菌孢子多糖组大鼠的粪菌组可以使代谢综合征大鼠的体重增长量、脂肪积累、肝脏脂质积累和炎症变性、胰岛素抵抗、血脂异常、机体炎症以及肠道损伤得到改善。
从图9中可以看出,与移植来自于代谢综合征大鼠的粪菌组来比,移植来自于冠突曲霉菌孢子多糖组大鼠的粪菌组可以使代谢综合征大鼠肠道菌群的多样性得到恢复;降低Firmicutes的相对丰度,提高Bacteroidetes的相对丰度以及Firmicutes/Bacteroidetes的比值;在属水平上,冠突曲霉菌孢子多糖能够显著提高代谢综合征大鼠体内Akkermansia,Alloprevotella,Bacteroides,Faecalibaculum,Rikenella,和Ruminococcaceae_UCG-005等有益菌的相对丰度,抑制Allobaculum,Blautia,Desulfovibrio,Escherichia-Shigella和Streptococcus等有害菌的相对丰度。
根据以上结果,我们得知,冠突曲霉菌孢子多糖的改善代谢综合征效果是基于对肠道菌群的改变而达到的。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对发明专利范围的限制。
应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (5)
1.一种冠突曲霉菌孢子多糖在制备改善代谢综合征的药物和功能性食品中的应用,其特征在于,所述的应用为改善代谢综合征导致的体重增加、脂肪积累、肝脏脂质积累和肝功能障碍、胰岛素抵抗、血脂紊乱、机体炎症和结肠屏障功能以及肠道菌群的失衡;其中,所述的冠突曲霉菌孢子多糖能够显著提高Akkermansia, Alloprevotella, Bacteroides,Faecalibaculum, Rikenella,和Ruminococcaceae_UCG-005有益菌的相对丰度,抑制Allobaculum, Blautia, Desulfovibrio, Escherichia-Shigella和Streptococcus有害菌的相对丰度;
所述的冠突曲霉菌孢子多糖由摩尔比为1:1.7:4.4:5.2的核糖、葡萄糖、半乳糖、甘露糖4种单糖组成;所述的冠突曲霉菌孢子多糖在所述改善代谢综合征的药物和功能性食品中,冠突曲霉菌孢子多糖的含量为0.1-99.5wt%;
所述的冠突曲霉菌孢子多糖的制备方法包括以下步骤:
S1、将冠突曲霉菌孢子破壁后进行干燥,研磨成粉末,按料液比为1g:20-50mL的比例与低共熔溶剂混匀,在70℃条件下反应30-55 min,随后抽滤取滤液;
S2、将滤液加入到乙醇溶液中,静置醇沉后离心,收集沉淀物;
S3、将沉淀物用蒸馏水溶解,再用Sevag法脱蛋白,最后将所得溶液经过透析后真空冻干,即得冠突曲霉菌孢子多糖;
其中,所述低共熔溶剂为氯化胆碱与草酸按摩尔比1:1混合后加热制得;
S1中所用的冠突曲霉菌孢子是通过将冠突曲霉菌接种于黑毛茶后进行散茶发花,然后将生长在茶叶中的冠突曲霉菌孢子经过振摇后,过筛去除杂质所得,所述冠突曲霉菌保藏编号为CCTCC NO:M20211325。
2.根据权利要求1所述的应用,其特征在于,
步骤S2中,
所述乙醇溶液为85-98 v%的乙醇水溶液。
3.根据权利要求1或2所述的应用,其特征在于,
步骤S2中,
所述静置醇沉具体为,置于2-6℃的冰箱中静置24-48h;
和/或,所述离心的转速为4000-8000rpm。
4.根据权利要求1所述的应用,其特征在于,
步骤S1中,
所述反应的时间为40min。
5.根据权利要求2所述的应用,其特征在于,
所述乙醇水溶液的加入体积为滤液的3-5倍。
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