CN114276942B - 谷胱甘肽酵母、产品的制备方法和应用 - Google Patents
谷胱甘肽酵母、产品的制备方法和应用 Download PDFInfo
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- CN114276942B CN114276942B CN202111670304.5A CN202111670304A CN114276942B CN 114276942 B CN114276942 B CN 114276942B CN 202111670304 A CN202111670304 A CN 202111670304A CN 114276942 B CN114276942 B CN 114276942B
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Classifications
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Abstract
本发明公开了一种谷胱甘肽酵母、产品的制备方法和应用。该生产方法包括:谷胱甘肽酵母菌株经多级种子培养后,转接到发酵罐进行培养,发酵得到富含谷胱甘肽的酵母,培养的过程中分三个阶段进行时长控制,包括菌体生长阶段、谷胱甘肽初期合成阶段和谷胱甘肽加速合成阶段;菌体生长阶段呼吸商值在1.0~1.2;谷胱甘肽初期合成阶段呼吸商值在0.7~1.0;谷胱甘肽加速合成阶段呼吸商值在0.5~0.7。本发明可以将酵母胞内谷胱甘肽含量提高到12~20%,属于较高水平,同时通过定量提取酵母胞内谷胱甘肽,可以开发出特定含量的谷胱甘肽营养酵母、谷胱甘肽酵母抽提物、谷胱甘肽酵母水解物产品,并能实现多种谷胱甘肽产品的联产。
Description
技术领域
本发明涉及生物技术及其应用领域,具体而言,涉及一种谷胱甘肽酵母、产品的制备方法和应用。
背景技术
谷胱甘肽是由谷氨酸、胱氨酸及甘氨酸组成的一种三肽,包括还原型谷胱甘肽和氧化型谷胱甘肽两种,其中还原型谷胱甘肽性质不稳定,很容易被氧化,因而是一种很好的抗氧化剂。同时还原型谷胱甘肽能激活多种酶,如体内的巯基(-SH)酶、辅酶等,从而促进糖类、脂肪及蛋白质代谢,也能影响细胞的代谢过程,是一种细胞内重要的调节代谢物质。谷胱甘肽目前已经被广泛应用于医药、面制品加工、食品、调味品、保健品、化妆品、动物营养、植物营养、微生物营养等领域,起到解毒、美白、抗氧化、提升风味、提高生产效率、提高机体免疫力等作用。
谷胱甘肽目前主要的生产方法有两种,一是利用筛选、驯化或基因改造后的酵母菌种,添加碳源、氮源、磷源、微量元素和前提氨基酸进行发酵,谷胱甘肽在酵母胞内得到富集;二是以谷氨酸、半胱氨酸、甘氨酸为底物,以γ-谷氨酰半胱氨酸合成酶、谷胱甘肽合成酶为催化剂,另外添加ATP、乙酸激酶、Mg2+、乙酰磷酸等能量辅助体系,酶法催化合成谷光甘肽。其中发酵法是当前谷胱甘肽主要生产方法,酶法合成属于新兴技术,但较难获得市场认可和原料药准入。
采用发酵法来生产谷胱甘肽近年来技术也不断升级,主要表现在二个方面,一是通过菌种选育、培养基优化和发酵过程精细化调控,谷胱甘肽含量不断提升,生产效率得到提高,生产成本大幅下降;二是随着生产成本的下降,谷胱甘肽产品形式和应用领域都得到极大拓展,更多的产品已经进入百姓的日常生活。
例如,中国专利CN101575578B公开一种酵母菌株、富含还原型谷胱甘肽的干酵母及其制备方法,包括以下步骤:酿酒酵母菌株→斜面菌种培养→培养瓶液体培养→旁氏罐液体培养→种子培养→发酵罐种子培养→发酵罐发酵培养→分离→干燥→包装。所述发酵工艺为:pH5.5~6.5、发酵温度28~32℃、通氧量0.5~1.5克O2/1克酵母*分钟、发酵时间20~22小时、酵母浓度170~250克/升。该发明制得的谷胱甘肽酵母中还原型谷胱甘肽的含量为3-5%。该专利所采用的发酵原料为常规酿酒酵母发酵原料,发酵过程没有进行精细化调控,谷胱甘肽含量水平低,同时产品形式为谷胱甘肽营养酵母,产品应用领域为大健康,比较单一。
中国专利CN113350229A公开一种富含谷胱甘肽酵母抽提物及制备方法和应用,通过将富含谷胱甘肽酵母原料经高压均质破壁得到酵母细胞壁乳,然后经复合酶解、灭酶处理和离心分离,取上清液得到富含谷胱甘肽酵母抽提物。其还原性谷胱甘肽含量达到5%以上,优选可以达到10%以上。该专利受限于酵母原料中谷胱甘肽水平,最终的抽提物产品中谷胱甘肽含量水平相对低,同时酵母经破壁、酶解、灭酶和离心分离后,轻相得到谷胱甘肽酵母抽提物,重相得到的是酵母细胞壁,酵母细胞壁属于谷胱甘肽酵母抽提物生产过程中的副产品,经济价值较低。
中国专利CN110959762A公开酵母水解物及其制备方法和应用,所述酵母水解物是通过酵母自溶,酶解,干燥后得到,产品中包含蛋白质、酸溶蛋白、酵母细胞壁、核酸和小肽,以酵母水解物干物质质量计,蛋白质的含量为40-45%,酸溶蛋白的含量为32-37%,小肽的含量为18-25%,酵母细胞壁的含量为25-35%,核酸的含量≥9%,酵母水解物的溶解率为50-60%,应用于动物养殖领域。该专利首次提供了酵母水解物的制备方法、产品和应用,但谷胱甘肽酵母水解物产品及应用未见相关专利。
因此,培养出谷胱甘肽含量水平高的酵母产品将具有很大的经济价值,并可以进一步拓宽后续产品的种类。
发明内容
本发明旨在提供一种谷胱甘肽酵母、产品的制备方法和应用,培养出谷胱甘肽含量水平高的酵母产品,同时通过定量提取酵母胞内谷胱甘肽,可以开发出特定含量的谷胱甘肽营养酵母、谷胱甘肽酵母抽提物、谷胱甘肽酵母水解物产品,并能实现多种谷胱甘肽产品的联产。
为了实现上述目的,根据本发明的一个方面,提供了一种富含谷胱甘肽的酵母的生产方法。该生产方法包括以下步骤:谷胱甘肽酵母菌株经多级种子培养后,转接到发酵罐进行培养,发酵得到富含谷胱甘肽的酵母,其中,在发酵罐中进行培养的过程中分三个阶段进行时长控制,三个阶段包括菌体生长阶段、谷胱甘肽初期合成阶段和谷胱甘肽加速合成阶段;菌体生长阶段呼吸商值在1.0~1.2;谷胱甘肽初期合成阶段呼吸商值在0.7~1.0;谷胱甘肽加速合成阶段呼吸商值在0.5~0.7。
进一步地,时长控制包括:在发酵罐中进行培养的时间一定的前提下,当产品要求的谷胱甘肽含量高时,缩短菌体生长阶段和谷胱甘肽初期合成阶段培养时间,延长谷胱甘肽加速合成阶段培养时间,控制单罐酵母产量,提升谷胱甘肽含量;当产品要求的谷胱甘肽含量低时,延长菌体生长阶段培养时间和谷胱甘肽初期合成阶段,缩短谷胱甘肽加速合成阶段培养时间,提高单罐酵母产量,谷胱甘肽含量相对降低。
进一步地,发酵罐中的发酵培养基包括淀粉水解糖30~40%、酵母抽提物1~3%、小麦蛋白胨1~3%、玉米浆2~10%、磷酸氢二铵0.2~0.5%、硫酸镁0.05~0.1%和硫酸锌0.02~0.04%,其中,水解糖的总糖浓度为28~32%。
进一步地,发酵罐中的发酵培养条件包括温度28~32℃,pH5.0~6.0,通气量为0.5~1.5vvm,培养时间为28~35小时,发酵结束时酵母细胞干重达到50g/L以上。
进一步地,谷胱甘肽酵母菌株的保藏号为CCTCC M205130。
进一步地,富含谷胱甘肽的酵母内谷胱甘肽含量为12~20%。
根据本发明的另一方面,提供了一种从酵母中定量提取谷胱甘肽的方法。该方法包括以下步骤:将通过上述任一种生产方法制备得到的富含谷胱甘肽的酵母发酵结束后,离心分离得到酵母乳,干物质控制10~20%;酵母乳升温到60~95℃,保温0~10分钟后降温,谷胱甘肽由酵母胞内释放到胞外;其中,通过控制提取温度和时间来控制谷胱甘肽的提取率。
根据本发明的再一方面,提供了一种富含谷胱甘肽的酵母。该酵母由上述任一种的富含谷胱甘肽的酵母的生产方法制备得到,富含谷胱甘肽的酵母内谷胱甘肽含量为12~20%。
根据本发明的又一方面,提供了一种谷胱甘肽酵母抽提物与谷胱甘肽营养酵母或谷胱甘肽酵母水解物联产的工艺。该工艺以上述富含谷胱甘肽的酵母为原料,按照本发明的上述方法提取谷胱甘肽,提取结束后离心分离,通过控制谷胱甘肽提取率使谷胱甘肽在轻相和重相中的分配比例在0-100%的任意值;其中,轻相作为谷胱甘肽酵母抽提物原料,重相作为谷胱甘肽营养酵母或谷胱甘肽酵母水解物原料进行谷胱甘肽酵母抽提物与谷胱甘肽营养酵母的联产,或谷胱甘肽酵母抽提物与谷胱甘肽酵母水解物的联产。
根据本发明的再一方面,提供了一种富含谷胱甘肽的营养酵母。该营养酵母由本发明的上述工艺制备得到,富含谷胱甘肽的营养酵母内谷胱甘肽含量为2~20%。
根据本发明的又一方面,提供了一种富含谷胱甘肽的酵母抽提物。该酵母抽提物由本发明的上述工艺制备得到,酵母抽提物中谷胱甘肽含量为5~50%,总氮含量≥8%,氨基酸态氮≥2%。
根据本发明的再一方面,提供了一种富含谷胱甘肽的酵母水解物。该酵母水解物由本发明的上述工艺制备得到,酵母水解物中谷胱甘肽含量为2~20%,蛋白质含量≥35%,酸溶蛋白含量≥30%,小肽含量≥15%。
根据本发明的再一方面,提供了一种富含谷胱甘肽的复配产品,该产品以谷胱甘肽营养酵母、谷胱甘肽酵母抽提物、谷胱甘肽酵母水解物为原料,取其中的两种或三种产品以任意比例混合后得到,复配产品中谷胱甘肽含量为2~50%。
根据本发明的又一方面,提供了上述富含谷胱甘肽的营养酵母、上述富含谷胱甘肽的酵母抽提物、上述富含谷胱甘肽的酵母水解物和/或上述富含谷胱甘肽的复配产品在食品、调味品、保健品、化妆品、动物营养品、植物营养品、微生物营养品制备方面的应用。
进一步地,食品为面制品时,应用为提升面团的延展性,促使面团成型;当食品为葡萄酒、果酒时,应用为防止果汁褐变、保持色泽或提升口感;应用为在保健品中作为抗氧化或美白组分;应用为在食品与调味品作为抗氧化、改善口感、提升风味的组分;应用为在化妆品作为清除自由基、抑制色素暗沉、美白抗氧化的组分;应用为在动物营养品、植物营养品或微生物营养品作为均衡营养、提高机体的消化吸收能力、促进生长、提高机体的免疫力或减少病害的组分。
应用本发明的技术方案,可以将酵母胞内谷胱甘肽含量提高到12~20%,属于较高水平,同时通过定量提取酵母胞内谷胱甘肽,可以开发出特定含量的谷胱甘肽产品,并能实现多种谷胱甘肽产品的联产,减少细胞壁这一副产品的产生,大幅降低谷胱甘肽产品的生产成本,推动谷胱甘肽营养酵母、谷胱甘肽酵母水解物和谷胱甘肽酵母抽提物产品质量升级和应用领域的拓展,能让更多谷胱甘肽产品进入百姓日常生活,经济和社会效益明显。
具体实施方式
需要说明的是,在不冲突的情况下,本申请中的实施例及实施例中的特征可以相互组合。下面将结合实施例来详细说明本发明。
根据本发明一种典型的实施方式,富含谷胱甘肽的酵母的生产方法。该方法包括以下步骤:谷胱甘肽酵母菌株经多级种子培养(即活化菌株,提高菌体量)后,转接到发酵罐进行培养,发酵得到所述富含谷胱甘肽的酵母,其中,在所述发酵罐中进行培养的过程中分三个阶段进行时长控制,所述三个阶段包括菌体生长阶段、谷胱甘肽初期合成阶段和谷胱甘肽加速合成阶段;所述菌体生长阶段呼吸商值在1.0~1.2;所述谷胱甘肽初期合成阶段呼吸商值在0.7~1.0;所述谷胱甘肽加速合成阶段呼吸商值在0.5~0.7。
本发明基于发酵过程尾气分析的三阶段控制策略为,通过在线实时监测菌体的生理状态,包括二氧化碳释放速率、氧气摄入速率和呼吸商。具体的,菌体生长阶段呼吸商值在1.0~1.2;谷胱甘肽初期合成阶段呼吸商值在0.7~1.0;谷胱甘肽加速合成阶段呼吸商值在0.5~0.7。进行三阶段培养的原因在于,实践中发现酵母胞内谷胱甘肽含量越高,比生长速率越慢,为了提高生产效率,第一阶段(菌体生长阶段)让酵母快速生长,第二阶段(谷胱甘肽初期合成阶段)酵母边生长边少量合成谷胱甘肽,第三阶段(谷胱甘肽加速合成阶段)酵母生长接近停滞,谷胱甘肽大量合成。实际生产过程中,在发酵时间一定的前提下,如果产品要求的谷胱甘肽含量高,可以适当缩短第一阶段和第二阶段培养时间,延长第三阶段培养时间,控制单罐酵母产量,谷胱甘肽含量可以得到提升;如果产品要求的谷胱甘肽含量低,可以适当延长第一和第二阶段培养时间,缩短第三阶段培养时间,提高单罐酵母产量,谷胱甘肽含量会降低。本申请中“谷胱甘肽含量高”和“谷胱甘肽含量低”都是相对于本发明技术方案中谷胱甘肽含量来说的,无需限定确定的值,本领域技术人员在本申请的发明思想和实施例的教导下,完全能够清楚本发明的对“谷胱甘肽含量高”和“谷胱甘肽含量低”的限定的含义,并根据实际情况实现本发明的技术方案。
另外,在实际生产过程中,为了方便操作,时长控制也可以通过细胞干重来体现,例如当细胞干重达到某预定值时则认为进入某个阶段,具体指标可以根据经验值进行确定。
优选的,发酵罐中的发酵培养基包括淀粉水解糖30~40%、酵母抽提物1~3%、小麦蛋白胨1~3%、玉米浆2~10%、磷酸氢二铵0.2~0.5%、硫酸镁0.05~0.1%和硫酸锌0.02~0.04%,其中,水解糖的总糖浓度为28~32%。小麦蛋白胨和玉米浆都属于市售产品,从市场购买即可,无特定指标限定。富含谷胱甘肽的酵母在此成分的发酵培养基中的发酵培养基利于提高谷胱甘肽的含量。
优选的,发酵罐中的发酵培养条件包括温度28~32℃,pH5.0~6.0,通气量为0.5~1.5vvm,培养时间为28~35小时,发酵结束时酵母细胞干重达到50g/L以上,实现高密度发酵。更优选的,谷胱甘肽酵母菌株的保藏号为CCTCC M205130(参见CN101575578B)。
在本发明一优选的实施方式中,谷胱甘肽酵母菌株经多级种子培养后,转接到发酵罐进行培养,在上述特定的培养基、培养条件和基于发酵过程微生物生理参数的三阶段控制策略下,发酵得到富含谷胱甘肽的酵母,该方法可使酵母胞内谷胱甘肽含量达到12-20%。
根据本发明一种典型的实施方式,本发明提供了一种从酵母中定量提取谷胱甘肽的方法。该方法不仅使终产品中的谷胱甘肽含量可控,满足不同客户对谷胱甘肽含量差异化需求,同时可以实现多种谷胱甘肽产品的联产,减少副产品的产生,进一步提升经济效益。具体方法为,谷胱甘肽酵母发酵结束后,离心分离得到干物质10~20%酵母乳,酵母乳升温到60-95℃,根据工艺需求保温0-10分钟后降温,谷胱甘肽由酵母胞内快速提取到胞外。生产过程中通过改变控制提取温度和时间来控制谷胱甘肽的提取率,使谷胱甘肽在轻相和重相中的分配比例在0-100%的任意值,从而得到不同含量的谷胱甘肽产品,同时可以实现谷胱甘肽酵母抽提物与谷胱甘肽营养酵母的联产,或者谷胱甘肽酵母抽提物与谷胱甘肽酵母水解物的联产,减少副产品酵母细胞壁的产生,降低产品生产成本。为提高提取效率,升温和降温设备优选板式换热器,具有换热效率高、热损失小、结构紧凑轻巧、占地面积小、使用寿命长等特点。
基于上述酵母中定量提取谷胱甘肽的方法,在实际生产过程中可以演化出多种工艺方案,包含但不限于以下工艺举例:
①在谷胱甘肽相关产品生产过程中,部分产品酶解时需要长时间的高温,会造成谷胱甘肽损失,为保证产品中谷胱甘肽含量,酵母乳在高温酶解前先进行谷胱甘肽提取,离心分离后,谷胱甘肽进入轻相并泵入物料缓储罐,重相按照常规工艺进行自溶和酶解,干燥前再将轻相和重相混合,可以减少谷胱甘肽在生产过程中的损失。
②在谷胱甘肽相关复配产品生产过程中,选取一批谷胱甘肽酵母乳进行谷胱甘肽提取,离心分离后,谷胱甘肽进入轻相并泵入物料缓储罐作为谷胱甘肽复配产品的生产原料,在相关产品生产中按需添加。可以节省干燥后再复配的生产成本。
根据本发明一种典型的实施方式,提供一种富含谷胱甘肽的酵母产品联产的方法。该方法包括以上述富含谷胱甘肽的酵母为原料,提取谷胱甘肽时控制提取率小于100%,使谷胱甘肽在轻相和重相中的分配比例在0-100%的任意值,提取结束后离心分离,轻相作为谷胱甘肽酵母抽提物原料,重相作为谷胱甘肽营养酵母或谷胱甘肽酵母水解物原料,实现谷胱甘肽酵母抽提物与谷胱甘肽营养酵母的联产,或谷胱甘肽酵母抽提物与谷胱甘肽酵母水解物的联产。
在本发明的一种典型的实施方式中,富含谷胱甘肽的营养酵母、富含谷胱甘肽酵母的水解物和富含谷胱甘肽的酵母抽提物产品详细生产工艺可参考CN101575578B、CN110959762A和CN113350229A的专利内容。
根据本发明一种典型的实施方式,还提供了富含谷胱甘肽的营养酵母、富含谷胱甘肽的酵母抽提物和富含谷胱甘肽的酵母水解物及复配产品。进一步地,富含谷胱甘肽的营养酵母中,谷胱甘肽含量为2-20%;富含谷胱甘肽的酵母水解物中谷胱甘肽含量为2-20%,蛋白质含量≥35%,酸溶蛋白含量≥30%,小肽含量≥15%;富含谷胱甘肽的酵母抽提物中谷胱甘肽含量为5-50%,总氮含量≥8%,氨基酸态氮≥2%;复配产品是指上述产品之间的复配组合实现特定功能的产品。
基于上述富含谷胱甘肽的营养酵母可以开发出2-20%谷胱甘肽营养酵母、2-20%谷胱甘肽酵母水解物和5-50%谷胱甘肽酵母抽提物及复配产品,广泛应用于食品、调味品、保健品,化妆品、动物营养、植物营养、微生物营养等领域,起到解毒、美白、抗氧化、提升风味、提高生产效率、提高机体免疫力等作用。进一步的,该谷胱甘肽产品及复配产品的使用包括但不限于以下领域:披萨、面饼等面制品制作,面粉中添加一定比例的谷胱甘肽营养酵母,可以大幅提升面团的延展性,促使快速成型,从而降低劳动强度,提高生产效率;葡萄酒、果酒等酿造过程中,可以有效防止果汁褐变,保持色泽,提升口感;作为保健品,可以起到抗氧化、美白等功效;食品与调味品中添加谷胱甘肽产品,可以抗氧化,改善口感,提升风味;化妆品中添加谷胱甘肽产品,可以清除自由基,抑制色素暗沉,达到美白抗氧化效果;在动物、植物、微生物营养领域,谷胱甘肽产品除了可以提供均衡营养,提高机体的消化吸收能力,促进生长外,还可以有效提高机体的免疫力,减少病害。
为了更好地理解和实施本发明,下面按照实施例、对比例详细说明本发明的技术方案,但本发明的保护内容不局限于以下实施例。实施例中谷胱甘肽营养酵母、谷胱甘肽酵母水解物和谷胱甘肽酵母抽提物产品详细生产工艺可参考CN101575578B、CN110959762A和CN113350229A的专利内容。
实施例1
谷胱甘肽酵母菌株(CCTCC M205130)经多级种子培养后,接种到发酵培养基中,水解糖原料的总糖浓度为30.1%,在发酵过程中流加,每小时的流加速率为发酵体积的0.1-1.6%。其余发酵原料用量包括酵母抽提物1%,小麦蛋白胨1%,玉米浆2%,磷酸氢二铵0.2%,硫酸镁0.05%,硫酸锌0.02%,余量的水。发酵温度28-30℃,pH5.0-5.5,通气量为0.5-1.5vvm,根据在线实时监测菌体的生理状态,包括二氧化碳释放速率、氧气摄入速率和呼吸商来反馈调控碳源和氮源的流加速率,当酵母细胞干重0-20g/L时,控制呼吸商1.0-1.2,当酵母细胞干重20-40g/L时,控制呼吸商0.7-1.0,当酵母细胞干重40g/L以上时,控制呼吸商0.5-0.7,培养28小时后,酵母细胞干重达到55.6g/L,谷胱甘肽含量达到酵母细胞干重的12.1%,蛋白质含量为45.3%。发酵结束后离心机分离,再用工艺水进行2次洗涤,最终得到干物质16.5%酵母乳。
实施例2
谷胱甘肽酵母菌株(CCTCC M205130)经多级种子培养后,接种到发酵培养基中,水解糖原料的总糖浓度为29.7%,在发酵过程中流加,每小时的流加速率为发酵体积的0.1-1.9%。其余发酵原料用量包括酵母抽提物3%,小麦蛋白胨3%,玉米浆10%,磷酸氢二铵0.5%,硫酸镁0.1%,硫酸锌0.04%,余量的水。发酵温度30-32℃,pH5.5-6.0,通气量为0.5-1.5vvm,根据在线实时监测菌体的生理状态,包括二氧化碳释放速率、氧气摄入速率和呼吸商来反馈调控碳源和氮源的流加速率,当酵母细胞干重0-15g/L时,控制呼吸商1.0-1.2,当酵母细胞干重15-30g/L时,控制呼吸商0.7-1.0,当酵母细胞干重30g/L以上时,控制呼吸商0.5-0.7,培养35小时后,酵母细胞干重达到50.3g/L,谷胱甘肽含量达到酵母细胞干重的20.2%,蛋白质含量为52.0%。发酵结束后离心机分离,并用工艺水进行2次洗涤,最终得到干物质14.9%酵母乳。
实施例3
按照实施例2重新制得谷胱甘肽酵母乳,经检测谷胱甘肽含量为20.8%,蛋白质含量为51.4%。按照谷胱甘肽营养酵母生产工艺得到谷胱甘肽营养酵母,经检测,谷胱甘肽含量为20.5%,蛋白质含量为51.5%。
取同一批次酵母乳,利用板式换热器将酵母乳升温到90-95度,并保温8分钟,再利用板式换热器降温到33度,5000rpm离心机分离,分别收集重相和轻相,轻相按照酵母抽提物生产工艺生产得到谷胱甘肽酵母抽提物,经检测谷胱甘肽含量为50.8%,总氮含量9.5%,氨基酸态氮3.7%;重相按照谷胱甘肽营养酵母工艺生产得到谷胱甘肽营养酵母,经检测谷胱甘肽含量为2.2%,蛋白质含量为48%。
取同一批次酵母乳,利用板式换热器将酵母乳升温到75-80度,并保温5分钟,再利用板式换热器降温到28度,5000rpm离心机分离,分别收集重相和轻相,轻相按照酵母抽提物生产工艺生产得到谷胱甘肽酵母抽提物,经检测谷胱甘肽含量为25.1%,总氮含量10.2%,氨基酸态氮5.1%;重相按照谷胱甘肽营养酵母工艺生产得到谷胱甘肽营养酵母,经检测谷胱甘肽含量为10.3%,蛋白质含量为50.3%。
实施例4
取实施例3中的谷胱甘肽酵母乳,按照谷胱甘肽酵母水解物生产工艺得到谷胱甘肽酵母水解物,经检测,谷胱甘肽含量为20.0%,蛋白质含量为50.7%,酸溶蛋白含量42.9%,小肽含量22%。
取实施例3中的谷胱甘肽酵母乳,利用板式换热器将酵母乳升温到90-95度,并保温8分钟,再利用板式换热器降温到32度,5000rpm离心机分离,分别收集重相和轻相,轻相按照酵母抽提物生产工艺生产得到谷胱甘肽酵母抽提物,经检测谷胱甘肽含量为50.1%,总氮含量10.5%,氨基酸态氮4.6%;重相按照酵母水解物工艺生产得到谷胱甘肽酵母水解物,经检测谷胱甘肽含量为2.4%,蛋白质含量为49.5%,酸溶蛋白含量40.2%,小肽含量19.0%。
取实施例3中的谷胱甘肽酵母乳,利用板式换热器将酵母乳升温到60-65度,并保温2分钟,再利用板式换热器降温到30度,5000rpm离心机分离,分别收集重相和轻相,轻相按照酵母抽提物生产工艺生产得到谷胱甘肽酵母抽提物,经检测谷胱甘肽含量为5.5%,总氮含量11.2%,氨基酸态氮5.5%;重相按照酵母水解物工艺生产得到谷胱甘肽酵母水解物,经检测谷胱甘肽含量为42.7%,蛋白质含量为50.3%。酸溶蛋白含量42.3%,小肽含量21.5%。
实施例5
将上述实施例中得到的谷胱甘肽产品,单独使用或相互复配后使用或与其它物质复配后使用,在不同的应用领域可以呈现不同的应用效果。
①披萨、面饼等面品制作时,面团中添加一定比例的谷胱甘肽营养酵母产品,可以大幅提升面团的延展性,促使面团快速成型,从而降低劳动强度,提高生产效率。实验室取300g白燕牌面包粉,加入纯化水195g制作面团作为对照组,实验组在对照组基础上按照面粉质量的0.5%添加2%含量的谷胱甘肽营养酵母制作面团,利用粉质仪FARINOGRAPH-E监测两种面团的揉混特性,实验组与对照组对比如表1所示。
表1:实验组和对照组粉质曲线特征参数比较
面团形成时间:一定范围内,面团形成时间越短,表示面筋量越少。
稳定性:一定范围内,稳定时间越短,说明面粉筋力越弱,韧性越差。
弱化度:一定范围内,弱化度越高,面筋越弱,面团越易变形。
②葡萄酒、果酒等酿造过程中添加一定比例的谷胱甘肽产品,可以增强其抗氧化能力,延缓酒的黄化、老化和褐变速度,提升产品的陈酿潜力和货架期,同时可以保护酒体香气,防止有益的挥发性硫醇的散失。实验室以酒精发酵完成后的葡萄酒原酒作为对照,实验组在原酒中加入25g/HL的谷胱甘肽产品(以谷胱甘肽计),实验组和对照组在同等条件下陈酿6个月后检测相关指标,对比如下表2。
表2
褐变指数测定:取一定量的葡萄酒至离心管中,12000r/min离心5min,离心后吸取上清液到比色皿中,以蒸馏水为对照,测定420nm下的吸光值来表示。
DPPH清除率测定:吸取2.0mL葡萄酒于试管中,加入0.2mmoL/L的DPPH溶液2mL混匀,避光反应30min,无水乙醇为参比,517nm测吸光值(A1)。测相同体积无水乙醇、DPPH溶液混匀的吸光值(A0)及2mL样品溶液和2mL无水乙醇混匀的吸光值(A2)。
DPPH清除率(%)=[1-(A1-A2)/A0]×100%。
总还原力测定:在10mL离心管中分别加入0.2mol/L pH 6.6的磷酸缓冲液2.5mL和葡萄酒0.2mL,加入2.5mL 1%铁氰化钾,混合均匀后于50℃反应20min。取出后加入1mL10%三氯乙酸终止反应,5000r/min离心10min。取上清液2.5mL,加入2.5mL蒸馏水和0.5mL0.1%FeCl3,混匀后静置10min,在700nm处检测吸光值。
③保健品中添加谷胱甘肽产品,可以起到抗氧化、解酒护肝等功效。应用实验内容可参见CN111920052A实施例。
④食品中添加一定比例的谷胱甘肽产品,可以充分发挥其抗氧化功效,起到稳定剂的作用,保持色泽,同时延长货架期。实验中在酸奶中按照2%比例添加10%含量的谷胱甘肽酵母抽提物,单位体积活菌数提高12.1%,货架期可以提高20%;调味品中添加一定比例的谷胱甘肽产品,作为小肽有呈味的功能,可以改善口感,提升风味。
⑤化妆品中添加一定比例的谷胱甘肽,可以清除自由基,抑制色素暗沉,达到美白抗氧化效果。应用实验内容可参见CN113350229A实施例。
⑥动物、植物、微生物营养领域,谷胱甘肽产品除了可以提供均衡营养,提高机体的消化吸收能力,促进生长外,还可以有效提高机体的抗逆性,减少病害。实验中在罗非鱼饲料中添加300ppm的谷胱甘肽产品(以谷胱甘肽计),罗非鱼的增重率(WGR)相对于对照组提高8.1%,蛋白质沉积率(PDR)提高了6.8%,经济效益明显;实验中在水稻幼苗培养基中添加1g/L的谷胱甘肽产品(以谷胱甘肽计),同等条件下培养10天,相对于对照组,初生根长度增加21.1%,侧生根数量增加了71.2%,说明一定浓度的谷胱甘肽有利于促进植物根系的生长;植物乳杆菌发酵培养基中添加5%谷胱甘肽酵母抽提物产品,同等条件下培养,相对于普通酵母抽提物培养基,植物乳杆菌发酵单位体积活菌数提高17.2%,冷冻干燥后活细胞率提高11.2%,说明谷胱甘肽有利于微生物增殖并保持活性,增强抗逆性。
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (2)
1.一种富含谷胱甘肽的酵母的生产方法,其特征在于,包括以下步骤:谷胱甘肽酵母菌株经多级种子培养后,转接到发酵罐进行培养,发酵得到所述富含谷胱甘肽的酵母,其中,在所述发酵罐中进行培养的过程中分三个阶段进行时长控制,所述三个阶段包括菌体生长阶段、谷胱甘肽初期合成阶段和谷胱甘肽加速合成阶段;所述菌体生长阶段呼吸商值在1.0~1.2;所述谷胱甘肽初期合成阶段呼吸商值在0.7~1.0;所述谷胱甘肽加速合成阶段呼吸商值在0.5~0.7;
所述发酵罐中的发酵培养基包括淀粉水解糖30~40%、酵母抽提物1-3%、小麦蛋白胨1~3%、玉米浆2~10%、磷酸氢二铵0.2~0.5%、硫酸镁0.05~0.1%和硫酸锌0.02~0.04%,其中,所述水解糖的总糖浓度为28~32%;
所述发酵罐中的发酵培养条件包括温度28~32℃,pH5.0~6.0,通气量为0.5~1.5vvm,培养时间为28~35小时,发酵结束时酵母细胞干重达到50g/L以上;
所述谷胱甘肽酵母菌株的保藏号为CCTCC M205130;
所述时长控制包括:在所述发酵罐中进行培养的时间一定的前提下,
当产品要求的谷胱甘肽含量高时,缩短所述菌体生长阶段和所述谷胱甘肽初期合成阶段培养时间,延长所述谷胱甘肽加速合成阶段培养时间,控制单罐酵母产量,提升谷胱甘肽含量;
当产品要求的谷胱甘肽含量低时,延长所述菌体生长阶段培养时间和所述谷胱甘肽初期合成阶段,缩短所述谷胱甘肽加速合成阶段培养时间,提高单罐酵母产量,谷胱甘肽含量相对降低。
2.根据权利要求1所述的生产方法,其特征在于,所述富含谷胱甘肽的酵母内谷胱甘肽含量为12~20%。
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