CN105309974B - 一种蛹虫草肽口服液的制备方法 - Google Patents
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Abstract
本发明公开了一种蛹虫草肽口服液的制备方法,包括以下步骤:蛹虫草的培养,将培养好的蛹虫草、培养基质原料粗粉碎后进行匀浆;湿法超微粉碎后利用纤维素酶、淀粉酶和果胶酶等进行水解,水解液进一步经过陶瓷膜过滤,滤过液经纳滤膜浓缩成一定比例的红棕色透明液体,灌装、灭菌后即可得到蛹虫草肽口服液。本发明产品生产周期短,产品生产过程中原料无损失,是一种产品产出率高、生产工艺简单、绿色环保的生产方法。此外,本发明蛹虫草肽口服液保健型功能食品可以作为日常保健长期饮用,还可以作为相关疾病辅助治疗的产品,拓宽了蛹虫草和蚕蛹的应用领域。
Description
技术领域
本发明属于食用菌加工技术领域,具体涉及一种蛹虫草肽口服液的制备方法。
背景技术
蛹虫草(Cordyceps militaris(L.)Link)又称为北虫草、北冬虫夏草、蛹草,隶属于真菌界子囊菌门(Ascomycota)、子囊菌纲(Ascomycetes)、粪壳菌亚纲(Sordariomycetidae)、肉座菌目(Hypocreales)、麦角菌科(Clavicipitaceae)、虫草属(Cordyceps),是虫草属真菌的模式种。至2002年世界上已发现虫草属真菌400多种,其中我国报道的有120种左右,蛹虫草在世界各地均有分布。蛹虫草是一类真菌侵染昆虫幼虫所形成的由菌核和子实体所组成的真菌结构。其宿主主要为鳞翅目的蚕蛾科、舟蛾科等。冬季其蛰居土里,真菌菌丝在幼虫体内吸收营养并生长,最后使幼虫死亡,到了夏季,真菌菌丝自鳞翅目昆虫蛹上生出子实体,形似草。它是最常见的、分布最广、具有较好药用价值的虫草菌。
蛹虫草分布广、生长快、易培养、含有多种活性成分,对人体具有多种生理保健功能。据研究资料显示,蛹虫草含有虫草素、虫草酸、虫草多糖、腺苷、甾醇等多种生物活性物质,具有增强免疫力、调节内分泌、抗疲劳、抗氧化、抗衰老、抗菌、抗肿瘤、抗缺氧、镇静作用、降血糖、保护肝肾及呼吸系统等多种生理功能。其化学成分和功能与著名的药用真菌冬虫夏草(Cordyceps sinensis(Berk.)Sacc)极其相似,某些活性成分如虫草素、核苷等物质含量还高于冬虫夏草。
近年来,科技工作者们在蛹虫草及其无性型的生物学特征;人工栽培、液体深层发酵培养;培养基组成及培养条件优化;菌丝体及代谢产物的化学成分检测、分离、提取、纯化及其功能;优良菌株选育、功能性食品研制、保健药物开发等方面进行了大量的研究,取得了一些可喜的成果。
对蛹虫草虽然已经进行了大量研究报道,也有许多产品面市,但未见桑蚕蛹虫草肽的相关研究及产品面市。本专利以桑蚕蛹虫草为原料,制备蛹虫草肽口服液。
发明内容
本发明的目的在于提供一种蛹虫草肽口服液的制备方法,旨在解决蛹虫草产品形式单调,缺乏具有滋肝益肾、降血压和延缓衰老功能口服液的问题。
本发明的另一目的在于提供制备获得的蛹虫草肽口服液。
本发明具体通过以下技术方案实现:
一种蛹虫草肽口服液的制备方法,包括以下步骤:
1)蛹虫草液体种子的制备
将液体种子培养基于121℃灭菌10min~15min,接入蛹虫草斜面菌种,接种量为2cm2菌丝块/瓶,22℃恒温震荡培养箱培养;
2)桑蚕蛹虫草的栽培
缫丝后的鲜蚕蛹,洗净直接破碎,过10目筛,将湿蚕蛹粉和小麦粒按照质量比为4:1的进行混合后,置入虫草栽培盆中,加入营养液,21℃灭菌20min~30min,冷却后接种蛹虫草液体种子培养,20℃恒温光照培养箱暗培养12~15d,待菌丝长满表层后,光暗交替培养7-10d后,固体培养基质长出大量橘黄色米粒大小的原基时转恒温培养,继续培养15d左右,直至子实体为5~8cm、头部膨大开始产孢时,将蛹虫草子实体和培养基质一同采收;
3)粗粉碎后加水、匀浆
将采收鲜蛹虫草子实体和基质直接粉碎过10目筛,粉碎后的物料中加入10倍质量的水,采用高速匀浆器进行匀浆,使固体颗粒过100目筛;
4)湿法高剪切超微粉碎
将匀浆后的原料置于高压均质机中进行均质,均质机压力为30~40Mpa;
5)纤维素酶、果胶酶、胃蛋白酶、中性蛋白酶、碱性蛋白酶
向均质好的料液内加入中性蛋白和胃蛋白酶,混匀后进行酶解,于50℃的温度下酶解1h后,继续用0.1M的NaOH调整pH为8,加入碱性蛋白酶,酶解2h,获得的酶解液加入柠檬酸溶液,调节匀浆后的料液pH值为5~6,对物料恒温振荡酶解1~2h,用0.1M的NaOH溶液调整料pH为7,升温至80℃并保温10min以破坏酶的活性;
6)获得蛹虫草肽口服液
过滤、浓缩后收集澄清滤液,灌装后经过喷淋式杀菌85℃条件下10min灭菌处理,冷却后即得红棕色的蛹虫草肽口服液。
进一步,步骤(1)中所述的液体种子培养基为:采用10%土豆汁、鲜牛奶、蚕蛹浆按1:1:2的体积比将三种液体混合复配、灭菌即得。
进一步,步骤(2)中营养液加入量为每1kg固体料中1~2L;接种量为每千克固体料接种蛹虫草液体种子30~100ml。所述的营养液为100ml的5%香蕉汁中加入1g复合氨基酸、0.05~0.5g黄嘌呤制得。
进一步,步骤(2)中所述的光暗交替培养条件为:白天在800Lux、22℃条件下光照12h;夜晚在15℃条件下暗培养12h;所述的恒温培养条件为:温度为19℃,每天光照16h,光强为1000Lux。
进一步,步骤(5)中中性蛋白和胃蛋白酶加酶量分别为底物的1%-2%和0.25%~1%,碱性蛋白酶加酶量为底物的1%。
进一步,步骤(5)中酶解所用酶及其用量为纤维素酶150~200U/g、淀粉酶1000~1200U/g、果胶酶100~120U/g。
以上制备方法制备所得到的蛹虫草肽口服液也在本发明的保护范围之内。
本发明的有益效果为:本发明产品生产周期短,按照此方法栽培的蛹虫草子实体中虫草素含量高,呈金黄色;产品生产过程中原料无损失,是一种产品产出率高、生产工艺简单、绿色环保的生产方法。此外,本发明蛹虫草肽口服液保健型功能食品可以作为日常保健长期饮用,还可以作为相关疾病辅助治疗的产品,拓宽了蛹虫草的应用领域,生产的蛹虫草肽口服液含有包括肽类、腺苷、虫草素、多糖、甾醇、类胡萝卜素、氨基酸、钙、铁等在内的多种生物活性成分,具有显著的降血压、滋肝益肾、延缓衰老的生物学功能。此外,本发明生产工艺简单,有利于满足消费者对便携型功能性饮料的消费需求。
附图说明
图1是本发明蛹虫草肽口服液的制备工艺流程。
具体实施方式
下面结合实施例对本发明做进一步的说明,以下所述,仅是对本发明的较佳实施例而已,并非对本发明做其他形式的限制,任何熟悉本专业的技术人员可能利用上述揭示的技术内容加以变更为同等变化的等效实施例。凡是未脱离本发明方案内容,依据本发明的技术实质对以下实施例所做的任何经过简单修改或等同变化,均落在本发明的保护范围内。
本发明公开了一种蛹虫草肽口服液的制备方法,包括以下步骤:蛹虫草的培养,将培养好的蛹虫草、培养基质原料;粗粉碎后进行匀浆;湿法超微粉碎后利用蛋白酶、纤维素酶、淀粉酶和果胶酶等进行水解,水解液进一步经过陶瓷膜过滤,滤过液经纳滤膜浓缩成一定比例的红棕色透明液体,灌装、灭菌后即可得到蛹虫草肽口服液。
实施例1
一种蛹虫草肽口服液的制备方法,其制备流程如图1所示,具体通过以下操作完成
S101 蛹虫草液体种子的制备
采用10%土豆汁、鲜牛奶、蚕蛹浆(鲜蚕蛹加等质量的水直接打磨成浆,过40目筛)按照1:1:2的体积比将三种液体混合复配、灭菌即得液体种子培养基,将液体种子培养基装入500ml三角瓶,每三角瓶装液量为200ml,于121℃灭菌10min~15min,接入蛹虫草斜面菌种,接种量为2cm2菌丝块/瓶,22℃恒温震荡培养箱培养3~5d。
S102 桑蚕蛹虫草的栽培
缫丝后的鲜蚕蛹,洗净直接破碎,过10目筛,将湿蚕蛹粉和小麦粒按照质量比为4:1的进行混合后,置入虫草栽培盆中,加入营养液,每1kg固体料中营养液加入量为1~2L,121℃灭菌20min~30min,冷却后接种培养;接种量为:每千克固体料中蛹虫草液体种子的接种量为30~100ml,固体培养基表层接种后,20℃恒温光照培养箱暗培养12~15d,待菌丝长满表层,即可见光培养,每天白天光照12h,光强为800Lux,温度为22℃,暗培养时温度为15℃,培养7-10d后,固体培养基质长出大量橘黄色米粒大小的原基时,转恒温培养,培养温度为19℃,每天光照16h,光强为1000Lux,继续培养15d左右,直至子实体为5~8cm、头部膨大开始产孢时,将蛹虫草子实体和培养基质一同采收。
其中营养液为100ml的5%香蕉汁中加入1g复合氨基酸、0.05~0.5g黄嘌呤制得。
S103 粗粉碎后加水、匀浆
将鲜蛹虫草子实体和基质直接粉碎(过10目筛),粉碎后的物料中加入10倍质量的水,采用高速匀浆器进行匀浆,使固体颗粒过100目筛。
S104 湿法高剪切超微粉碎
将匀浆后的原料置于高压均质机中进行均质,均质机压力为30~40Mpa。
S105 纤维素酶、淀粉酶、果胶酶水解
向均质好的料液内加入中性蛋白和胃蛋白酶,混匀后进行酶解,中性蛋白和胃蛋白酶加酶量分别为底物的1%-2%和0.25%~1%,于50℃的温度条件下酶解1h后,继续用0.1M的NaOH调整pH为8,加入碱性蛋白酶,加酶量为底物的1%,酶解2h,获得的酶解液加入柠檬酸溶液,调节匀浆后的料液pH值处于5~6之间,酶解所用酶及用量为纤维素酶150~200U/g、淀粉酶1000~1200U/g、果胶酶100~120U/g,对物料恒温振荡酶解1~2h,用0.1M的NaOH溶液调整料pH为7,升温至80℃并保温10min以破坏酶的活性。
S106 过滤、浓缩、灌装
灭活后的料液采用压滤机去除固形物,滤过液经过0.45um的陶瓷膜过滤澄清处理,收集澄清滤过液再经截留分子量为200的纳滤膜过滤进行纯化、浓缩后,收集截留液定容(使每滤液10ml中活性物质含量折合3g蛹虫草子实体和基质混合物),灌装后经过喷淋式杀菌85℃条件下10min灭菌处理,冷却后即得红棕色的蛹虫草肽口服液。
Claims (4)
1.一种蛹虫草肽口服液的制备方法,其特征在于,包括以下步骤:
1)蛹虫草液体种子的制备
将液体种子培养基于121℃灭菌10min~15min,接入蛹虫草斜面菌种,接种量为2cm2菌丝块/瓶,22℃恒温震荡培养箱培养;
2)桑蚕蛹虫草的栽培
缫丝后的鲜蚕蛹,洗净直接破碎,过10目筛,将湿蚕蛹粉和小麦粒按照质量比为4:1的进行混合后,置入虫草栽培盆中,加入营养液,21℃灭菌20min~30min,冷却后接种蛹虫草液体种子培养,20℃恒温光照培养箱暗培养12~15d,待菌丝长满表层后,光暗交替培养7-10d后,固体培养基质长出大量橘黄色米粒大小的原基时转恒温培养,继续培养15d左右,直至子实体为5~8cm、头部膨大开始产孢时,将蛹虫草子实体和培养基质一同采收;
3)粗粉碎后加水、匀浆
将采收鲜蛹虫草子实体和基质直接粉碎过10目筛,粉碎后的物料中加入10倍质量的水,采用高速匀浆器进行匀浆,使固体颗粒过100目筛;
4)湿法高剪切超微粉碎
将匀浆后的原料置于高压均质机中进行均质,均质机压力为30~40Mpa;
5)纤维素酶、淀粉酶、果胶酶水解
向均质好的料液内加入中性蛋白和胃蛋白酶,混匀后进行酶解,于50℃的温度条件下酶解1h后,继续用0.1M的NaOH调整pH为8,加入碱性蛋白酶,酶解2h,获得的酶解液加入柠檬酸溶液,调节匀浆后的料液pH值为5~6之间,对物料恒温振荡酶解1~2h,用0.1M的NaOH溶液调整料pH为7,升温至80℃并保温10min以破坏酶的活性;
6)获得蛹虫草肽口服液
过滤、浓缩后收集澄清滤液,灌装后经过喷淋式杀菌85℃条件下10min灭菌处理,冷却后即得红棕色的蛹虫草肽口服液;
步骤(1)中所述的液体种子培养基为:采用10%土豆汁、鲜牛奶、蚕蛹浆按1:1:2的体积比将三种液体混合复配、灭菌即得;
步骤(2)中营养液加入量为每1kg固体料中1~2L;接种量为每千克固体料接种蛹虫草液体种子30~100ml;所述的营养液为100ml的5%香蕉汁中加入1g复合氨基酸、0.05~0.5g黄嘌呤制得;
步骤(2)中所述的光暗交替培养条件为:白天在800Lux、22℃条件下光照12h;夜晚在15℃条件下暗培养12h;所述的恒温培养条件为:温度为19℃,每天光照16h,光强为1000Lux。
2.根据权利要求1所述的一种蛹虫草肽口服液的制备方法,其特征在于,步骤(5)中中性蛋白和胃蛋白酶加酶量分别为底物的1%-2%和0.25%~1%,碱性蛋白酶加酶量为底物的1%。
3.根据权利要求1所述的一种蛹虫草肽口服液的制备方法,其特征在于,步骤(5)中中酶解所用酶及用量为纤维素酶150~200U/g、淀粉酶1000~1200U/g、果胶酶100~120U/g。
4.权利要求1~3中任一项所述的制备方法制备所得的蛹虫草肽口服液。
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