CN113303419A - 一种药食用真菌混合发酵菌丝体冲调粉的制备工艺 - Google Patents
一种药食用真菌混合发酵菌丝体冲调粉的制备工艺 Download PDFInfo
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Abstract
本发明公开了一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,涉及药食用真菌混合发酵技术领域,具体为一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,包括以下步骤:(1)采用发酵罐液态深层发酵的方法培养多种药食用真菌菌丝体,培养温度控制在20~25℃,搅拌转速100~500rpm,培养时间4~7天;(2)发酵结束后加入纯化水稀释后进行膜微滤分离和菌丝体超滤浓缩;(3)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH并加热使水解酶失活。本发明通过药食用真菌的混合培养获得富含多糖、萜类、核苷类和甾醇类等生物活性物质,克服了单一药食用真菌活性成分不丰富或者功效不突出的弱点。
Description
技术领域
本发明涉及药食用真菌混合发酵技术领域,具体为一种药食用真菌混合发酵菌丝体冲调粉的制备工艺。
背景技术
药食用真菌没有统一规范的定义,泛指既对人体疾病具有预防、保健和治疗功能,又可日常使用的微生物真菌。药食用真菌在我国具有悠久的药用和食用历史,随着现代生物科技和医学研究的进步,药食用真菌在疾病预防、保健和治疗方面的功效已被广泛证实,并由此产生了许多知名的临床药品和保健产品,例如具有调节免疫而达到抗肿瘤作用的香菇多糖;具有补益肺肾、秘精益气的发酵虫草粉(Cs-4,金水宝/百令胶囊);具有保肝、抗癌和调节免疫力的灵芝孢子粉。持此之外,许多珍稀药食用真菌都具有疾病的预防、保健和治疗功效,包括有冬虫夏草、蜜环菌、灰树花、羊肚菌、金耳、木耳、桑黄、姬松茸、云芝、樟芝、松口蘑、茯苓、猪苓、蝉花、雷丸、竹荪和猴头菇等。现代研究表明,药食用真菌含有的主要生理活性物质有多糖、萜类物质、核苷类物质、甾醇类物质和糖蛋白等。
现代生物学研究已表明,药食用真菌的人工发酵菌丝体(固态和液态)的生理成分含量和功效均与子实体相当,甚至优于子实体。张劲松等研究发现灵芝菌丝体提取物在高浓度时的免疫活性显著优于子实体提取物(菌物学报,2004,23(1):85-92)。回晶等研究表明桑黄菌丝体中的粗蛋白、必需氨基酸、脂肪和总糖含量明显高于子实体(特产研究,2009,31(002):59-61)。朱春玉等研究证实人工培养的桦褐孔菌菌丝体与子实体具有相似的药用价值,其中菌丝体三萜化合物对人胃癌细胞株MGC-803的抑制率达到38.27%(食品科学,2012,33(015):161-165)。
目前围绕药食用菌丝体或子实体的研究和产品开发绝大多数是单一菌株,也就是纯种培养,未见有多菌株混合培养的相关报道。专利CN112471377A公开了一种护肤养颜食用菌冲饮粉的制备方法,经食用菌子实体浸泡处理、真空干燥处理、糖衣包覆处理、高温高压处理和深冷粉碎处理获得了具有消除人体自由基和抗氧化功能的冲调粉。但该发明并未涉及多种食用菌混合处理,也未涉及菌丝体的工艺研究。
发明内容
本发明以多种药食用真菌为研究对象,采用2种及2种以上的液体混合培养方法,经液体深层发酵、膜过滤、酶解、流化床制粒、复配和包装步骤制备了富含多糖类、萜类、核苷类和甾醇类物质的药食用真菌菌丝体冲调粉。
为实现以上目的,本发明通过以下技术方案予以实现:一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,包括以下步骤:
(1)采用发酵罐液态深层发酵的方法培养多种药食用真菌菌丝体,培养温度控制在20~25℃,搅拌转速100~500rpm,培养时间4~7天;
(2)发酵结束后加入纯化水稀释后进行膜微滤分离和菌丝体超滤浓缩;
(3)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH并加热使水解酶失活;
(4)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;
(5)将制得的冲调粉进行质量检测和评价。
可选的,所述步骤(1)中的多种药食用真菌可以是灵芝、虫草、金耳、灰树花、猴头菇、香菇、蜜环菌、桑黄中任意二种或二种以上组合。
可选的,所述步骤(2)中的微滤膜可以是无机陶瓷膜、有机醋酸纤维素膜、硝酸纤维素膜、聚醚砜膜中的任何一种。
可选的,所述步骤(2)中所用的超滤膜可以是醋酸纤维素膜、聚醚砜膜、聚丙烯晴膜、聚偏氟乙烯膜、聚砜膜中的任何一种。
可选的,所述步骤(2)中超滤膜截留分子量范围为5K~30K。
可选的,所述步骤(3)中使用的水解酶可以是糖化酶、几丁质酶、纤维素酶、葡聚糖酶、蛋白酶中任何一种或多种酶复合使用。
可选的,所述步骤(3)中灭活酶的pH范围为7~9.5,加热温度为50~65℃。
可选的,所述步骤(4)中流化床制粒的进风温度为45~55℃,风速频率为8~25Hz,物料温度为25~35℃,浓缩液泵速为16~22ml/min,喷枪压力0.1~0.3Mpa。
本发明提供了一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,具备以下有益效果:
本发明的有益效果是通过药食用真菌的混合培养获得富含多糖、萜类、核苷类和甾醇类等生物活性物质,克服了单一药食用真菌活性成分不丰富或者功效不突出的弱点,为人们的健康和保健提供了更好的选择。
具体实施方式
下面,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。
实施案例一
本发明提供一种技术方案:一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,具体包括以下步骤:
(1)采用改良的马丁固体培养基分别制备不同的药食用真菌母种,这些药食用真菌可以是灵芝、虫草、金耳、灰树花、猴头菇、香菇、蜜环菌、桑黄中的任意二种或二种以上组合,培养温度23~25℃,相对湿度50~65%,培养时间5~7天;
(2)将母种斜面分别接种改良的PDA液体培养基,接种量5~8平方厘米,培养温度23~25℃,转速220rpm,培养时间72h;
(3)种子培养成熟后按照1~3%的接种量分别将不同食用菌种子液接种到10L发酵罐中进行发酵,发酵液中碳源为葡萄糖、麦芽糊精、土豆浸出物,氮源为酵母浸粉和大豆粉,并在培养基中添加有机羧酸草酸、柠檬酸和丙酮酸混合物,发酵通气量为0.4~1.2vvm,发酵温度为25℃,搅拌速度150~400rpm,发酵时间为144~168h,并经HPLC检测,确保菌丝体中所含活性成分总多糖、总萜、核苷类和甾醇中的任一类别含量不低于5mg/g;
(4)发酵结束后,发酵液经陶瓷膜微滤和超滤进行菌丝体收集和浓缩,陶瓷膜微滤的孔径范围为800nm~2μm,优选地孔径大小为0.8~1.5μm;陶瓷膜超滤膜截留分子量范围为5~30KD,优选地截留分子量范围为10~20KD;(5)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH并加热使水解酶失活;水解酶可以是糖化酶、几丁质酶、纤维素酶、葡聚糖酶、蛋白酶中任何一种或多种酶复合使用;
(5)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;流化床制粒条件为:进风温度为45~55℃,风速频率为8~25Hz,物料温度为25~35℃,浓缩液泵速为16~22ml/min,喷枪压力0.1~0.3Mpa;复配所用矫味剂可以是蔗糖、麦芽糖醇、甘露醇、甜菊糖苷、枸橼酸、抗坏血酸中的一种或多种复合,添加量为0.5~5%。
实施案例二
一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,具体包括以下步骤:
(1)采用改良的马丁固体培养基分别制备不同的药食用真菌母种,这些药食用真菌是灵芝、虫草、金耳和猴头菇四种组合,培养温度23℃,相对湿度50%,培养时间7天;
(2)将母种斜面分别接种改良的PDA液体培养基,接种量5平方厘米,培养温度23℃,转速220rpm,培养时间72h;
(3)种子培养成熟后按照1%的接种量分别将不同食用菌种子液接种到10L发酵罐中进行发酵,发酵液中碳源为葡萄糖2%、麦芽糊精4%,氮源为酵母浸粉1.5%和大豆粉2%,并在培养基中添加有机羧酸草酸0.15%、柠檬酸0.1%和丙酮酸0.05%混合物,发酵通气量为0.4~1.2vvm,发酵温度为25℃,搅拌速度150~400rpm,发酵时间为168h,并经HPLC检测,确保发酵混合菌丝中所含活性成分总萜、总黄酮、核苷类和甾醇中的任一类别含量不低于5mg/g;
(4)发酵结束后,发酵液经陶瓷膜微滤和超滤进行菌丝体收集和浓缩,陶瓷膜微滤的孔径范围为0.8μm;陶瓷膜超滤膜截留分子量范围为10KD;
(5)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH至7.0,并加热至50℃使水解酶失活;水解酶是糖化酶、几丁质酶和纤维素酶复合酶,加酶量200U/g;
(6)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;流化床制粒条件为:进风温度为45℃,风速频率为8~25Hz,物料温度为25~30℃,浓缩液泵速为16ml/min,喷枪压力0.2Mpa;复配所用矫味剂是蔗糖和麦芽糖醇复合物,添加量为1%。
实施案例三
一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,具体包括以下步骤:
(1)采用改良的马丁固体培养基分别制备不同的药食用真菌母种,这些药食用真菌是灵芝、虫草和灰树花三种组合,培养温度25℃,相对湿度65%,培养时间7天;
(2)将母种斜面分别接种改良的PDA液体培养基,接种量8平方厘米,培养温度25℃,转速220rpm,培养时间72h;
(3)种子培养成熟后按照3%的接种量分别将不同食用菌种子液接种到10L发酵罐中进行发酵,发酵液中碳源为葡萄糖2%、麦芽糊精4%,氮源为酵母浸粉1.5%和大豆粉2%,并在培养基中添加有机羧酸草酸0.15%、柠檬酸0.1%和丙酮酸0.05%混合物,发酵通气量为0.4~1.2vvm,发酵温度为25℃,搅拌速度150~400rpm,发酵时间为168h,并经HPLC检测,确保发酵液中所含活性成分总萜、总黄酮、核苷类和甾醇中的任一类别含量不低于5mg/g;
(4)发酵结束后,发酵液经陶瓷膜微滤和超滤进行菌丝体收集和浓缩,陶瓷膜微滤的孔径为1.5μm;陶瓷膜超滤膜截留分子量为10KD;
(5)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH至9.5,并加热至60℃使水解酶失活;水解酶是糖化酶、几丁质酶、纤维素酶和蛋白酶四种复合酶,加酶量180U/g;
(6)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;流化床制粒条件为:进风温度为50℃,风速频率为10~20Hz,物料温度为28~32℃,浓缩液泵速为20ml/min,喷枪压力0.3Mpa;复配所用矫味剂是蔗糖和甜菊糖苷复合物,添加量为1.5%。
实施案例四
一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,具体包括以下步骤:
(1)采用改良的马丁固体培养基分别制备不同的药食用真菌母种,这些药食用真菌是灵芝和虫草两种组合,培养温度24℃,相对湿度65%,培养时间7天;
(2)将母种斜面分别接种改良的PDA液体培养基,接种量8平方厘米,培养温度24℃,转速220rpm,培养时间72h;
(3)种子培养成熟后按照3%的接种量分别将不同食用菌种子液接种到10L发酵罐中进行发酵,发酵液中碳源为葡萄糖2%、麦芽糊精4%,氮源为酵母浸粉1.5%和大豆粉2%,并在培养基中添加有机羧酸草酸0.15%、柠檬酸0.1%和丙酮酸0.05%混合物,发酵通气量为0.4~1.2vvm,发酵温度为25℃,搅拌速度150~400rpm,发酵时间为168h,并经HPLC检测,确保发酵液中所含活性成分总萜、总黄酮、核苷类和甾醇中的任一类别含量不低于5mg/g;
(4)发酵结束后,发酵液经陶瓷膜微滤和超滤进行菌丝体收集和浓缩,陶瓷膜微滤的孔径为1.0μm;陶瓷膜超滤膜截留分子量为15KD;
(5)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH至8.0,并加热至65℃使水解酶失活;水解酶是几丁质酶和纤维素酶复合酶,加酶量100U/g;
(6)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;流化床制粒条件为:进风温度为50℃,风速频率为10~20Hz,物料温度为28~30℃,浓缩液泵速为18ml/min,喷枪压力0.25Mpa;复配所用矫味剂可以是蔗糖和枸橼酸复合物,添加量为0.8%。
实施案例五
一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,具体包括以下步骤:
(1)采用改良的马丁固体培养基分别制备不同的药食用真菌母种,这些药食用真菌是虫草、猴头菇、香菇和蜜环菌四种组合,培养温度25℃,相对湿度55%,培养时间7天;
(2)将母种斜面分别接种改良的PDA液体培养基,接种量5~8平方厘米,培养温度25℃,转速220rpm,培养时间72h;
(3)种子培养成熟后按照3%的接种量分别将不同食用菌种子液接种到10L发酵罐中进行发酵,发酵液中碳源为葡萄糖2%、麦芽糊精4%,氮源为酵母浸粉1.5%和大豆粉2%,并在培养基中添加有机羧酸草酸0.15%、柠檬酸0.1%和丙酮酸0.05%混合物,发酵通气量为0.4~1.2vvm,发酵温度为25℃,搅拌速度150~400rpm,发酵时间为168h,并经HPLC检测,确保发酵液中所含活性成分总萜、总黄酮、核苷类和甾醇中的任一类别含量不低于5mg/g;
(4)发酵结束后,发酵液经陶瓷膜微滤和超滤进行菌丝体收集和浓缩,陶瓷膜微滤的孔径为1.2μm;陶瓷膜超滤膜截留分子量为25KD;
(5)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH至7.5并加热至65℃使水解酶失活;水解酶是糖化酶、几丁质酶和纤维素酶复合酶,加酶量250U/g;
(6)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;流化床制粒条件为:进风温度为45℃,风速频率为8~15Hz,物料温度为25~27℃,浓缩液泵速为16ml/min,喷枪压力0.2Mpa;复配所用矫味剂可以是蔗糖和枸橼酸复合物,添加量为2.5%。
实施案例六
一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,具体包括以下步骤:
(1)采用改良的马丁固体培养基分别制备不同的药食用真菌母种,这些药食用真菌是灵芝、虫草、金耳、灰树花、猴头菇、香菇、蜜环菌、桑黄八种组合,培养温度25℃,相对湿度60%,培养时间7天;
(2)将母种斜面分别接种改良的PDA液体培养基,接种量8平方厘米,培养温度25℃,转速220rpm,培养时间72h;
(3)种子培养成熟后按照3%的接种量分别将不同食用菌种子液接种到10L发酵罐中进行发酵,发酵液中碳源为葡萄糖2%、麦芽糊精4%,氮源为酵母浸粉1.5%和大豆粉2%,并在培养基中添加有机羧酸草酸0.15%、柠檬酸0.1%和丙酮酸0.05%混合物,发酵通气量为0.4~1.2vvm,发酵温度为25℃,搅拌速度150~400rpm,发酵时间为144h,并经HPLC检测,确保发酵液中所含活性成分总萜、总黄酮、核苷类和甾醇中的任一类别含量不低于5mg/g;
(4)发酵结束后,发酵液经陶瓷膜微滤和超滤进行菌丝体收集和浓缩,陶瓷膜微滤的孔径为800nm;陶瓷膜超滤膜截留分子量为20KD;
(5)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH至8.0并加热至65℃使水解酶失活;水解酶可以是糖化酶、几丁质酶、纤维素酶、葡聚糖酶和蛋白酶复合酶,加酶量200U/g;
(6)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;流化床制粒条件为:进风温度为48℃,风速频率为10~16Hz,物料温度为26~29℃,浓缩液泵速为18ml/min,喷枪压力0.2Mpa;复配所用矫味剂可以是蔗糖、麦芽糖醇和枸橼酸复合物,添加量为2.5%。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (8)
1.一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,包括以下步骤:
(1)采用发酵罐液态深层发酵的方法培养多种药食用真菌菌丝体,培养温度控制在20~25℃,搅拌转速100~500rpm,培养时间4~7天;
(2)发酵结束后加入纯化水稀释后进行膜微滤分离和菌丝体超滤浓缩;
(3)将浓缩的菌丝体进行胶体磨研磨,研磨后加入水解酶进行酶解处理,酶解处理后调节pH并加热使水解酶失活;
(4)将处理后的菌丝体进行流化床制粒干燥,并复配适宜的矫味剂,制得冲调剂成品;
(5)将制得的冲调粉进行质量检测和评价。
2.根据权利要求1所述的一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,其特征在于:所述步骤(1)中的多种药食用真菌可以是灵芝、虫草、金耳、灰树花、猴头菇、香菇、蜜环菌、桑黄中任意二种或二种以上组合。
3.根据权利要求1所述的一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,其特征在于:所述步骤(2)中的微滤膜可以是无机陶瓷膜、有机醋酸纤维素膜、硝酸纤维素膜、聚醚砜膜中的任何一种。
4.根据权利要求1所述的一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,其特征在于:所述步骤(2)中所用的超滤膜可以是醋酸纤维素膜、聚醚砜膜、聚丙烯晴膜、聚偏氟乙烯膜、聚砜膜中的任何一种。
5.根据权利要求1所述的一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,其特征在于:所述步骤(2)中超滤膜截留分子量范围为5K~30K。
6.根据权利要求1所述的一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,其特征在于:所述步骤(3)中使用的水解酶可以是糖化酶、几丁质酶、纤维素酶、葡聚糖酶、蛋白酶中任何一种或多种酶复合使用。
7.根据权利要求1所述的一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,其特征在于:所述步骤(3)中灭活酶的pH范围为7~9.5,加热温度为50~65℃。
8.根据权利要求1所述的一种药食用真菌混合发酵菌丝体冲调粉的制备工艺,其特征在于:所述步骤(4)中流化床制粒的进风温度为45~55℃,风速频率为8~25Hz,物料温度为25~35℃,浓缩液泵速为16~22ml/min,喷枪压力0.1~0.3Mpa。
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