CN112913999A - 一种枸杞红枣复合发酵式功能饮品及其制备方法 - Google Patents
一种枸杞红枣复合发酵式功能饮品及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种枸杞红枣复合发酵式功能饮品及其制备方法,所述枸杞红枣复合发酵式功能饮品的制备方法包括以下步骤:将枸杞与无菌水等质量混合得到枸杞发酵基质,向枸杞发酵基质中接种纳豆芽孢杆菌的菌液,进行固态发酵;向固态发酵结束后的基质中加入无菌水,进行液态发酵;将液态发酵后的发酵液过滤,收集滤液,滤液离心后得到枸杞提取液;将枸杞提取液与红枣粉末、木糖醇粉末、水混合均匀,经过滤、杀菌、包装,即可得到所述枸杞红枣复合发酵式功能饮品,本发明公开的枸杞红枣复合发酵式功能饮品的原料组成虽然简单,但是却具有较高的营养价值和饮用价值,其具有溶栓、抗氧化、抗衰老、美容养颜等功效。
Description
技术领域
本发明属于饮品加工领域,具体涉及一种枸杞红枣复合发酵式功能饮品及其制备方法。
背景技术
心血管疾病(CVDs)是全球死亡的主要原因,其主要原因是血栓形成。目前,临床应用的溶栓药物(如尿激酶、链激酶、重组型纤溶酶原激活剂等)存在半衰期短、用药量大、对纤维蛋白的溶解不具有选择性,易导致内出血或引起超敏反应等副作用。
纳豆激酶(Nattokinase,NK)具有显著的溶解血栓的能力,其溶解血凝块的能力为血纤溶酶的4倍,NK具有安全、成本低、易口服等特点,被认为是治疗血栓的理想功能性食品。目前公开的纳豆激酶的制备方法均是以大豆作为固体基质进行固态发酵24h,由此得到的纳豆激酶主要是纳豆及其酵素,存在异味,由其制备成的功能性饮品也不适合中国人的口味。
枸杞是药食同源物质,枸杞、红枣具有药用价值及其去味的作用,其口感很受国人喜爱。但枸杞传统食用方式以干果为主,市场上出现的枸杞类发酵功能饮料中的枸杞多是应用酵母菌和乳酸菌进行简单发酵,其发酵周期较长,且发酵液中多为酵素,饮用价值不高。
发明内容
本发明的目的在于提供一种枸杞红枣复合发酵式功能饮品及其制备方法,以枸杞发酵液、红枣、木糖醇为主要原料,基于固/液体顺序发酵与生物催化共反应法,制备成的一款饮料,其富含蛋白质、多肽、枸杞多糖、纳豆激酶等功能组分,是一种多功能复合饮品。
本发明采取的技术方案为:
一种枸杞红枣复合发酵式功能饮品的制备方法,包括以下步骤:
(1)将枸杞与无菌水等质量混合得到枸杞发酵基质,向枸杞发酵基质中接种纳豆芽孢杆菌的菌液,进行固态发酵;
(2)向固态发酵结束后的基质中加入无菌水,进行液态发酵;
(3)将液态发酵后的发酵液过滤,收集滤液,滤液离心后得到枸杞发酵液;
(4)将枸杞发酵液与红枣粉末、木糖醇粉末、水混合均匀,经过滤、杀菌、包装,即可得到所述枸杞红枣复合发酵式功能饮品。
进一步地,步骤(4)中,所述枸杞红枣复合发酵式功能饮品中各原料的质量百分比分别为:枸杞提取液30%~50%、红枣粉末10%~20%、木糖醇粉末5%~10%,其余为水。。
步骤(1)中,所述固态发酵的条件为35-40℃,150-220r/min摇床培养4-12h,优选为37℃,200r/min摇床培养6h。
步骤(2)中,所述液态发酵的条件为35-40℃,150-220r/min摇床培养4-12h,优选为37℃,200r/min摇床培养6h。
所述纳豆芽孢杆菌的菌液浓度为1×107~1×108CFU/mL,菌液的接种量为枸杞发酵基质质量的5%。
所述纳豆芽孢杆菌的菌液的制备方法为:将纳豆芽孢杆菌于种子培养基中活化,再用酪蛋白平板筛选透明圈大的菌落,挑取筛选出的菌落至种子培养基中扩大培养。
所述种子培养基的组成为:胰蛋白胨6~8g、葡萄糖10~12g、牛肉浸膏3~5g、蒸馏水600mL、pH为7.2。
步骤(2)中的无菌水的加入量为枸杞质量的2.5~4倍。
所述纳豆芽孢杆菌为纳豆枯草芽孢杆菌。
本发明提供的一种枸杞红枣复合发酵式功能饮品,采用上述制备方法制备得到。
本发明提供的枸杞红枣复合发酵式功能饮品的制备方法中,以纳豆芽孢杆菌为发酵菌种,通过固态发酵预处理软化分解营养基质,累积次级代谢产物,促进液体发酵时微生物代谢吸收和酶促催化反应,生产NK和蛋白酶,蛋白酶又能降解枸杞中糖蛋白复合物中的蛋白质,形成多肽,便于微生物代谢吸收,制备出富含NK、蛋白质、多肽和枸杞多糖等多种保健功能组分并存的枸杞发酵液,整个制备过程不超过24h;即上述制备方法中,固态发酵过程中所积累的胞外蛋白酶等酶类,可加速液态发酵条件下的生物催化反应过程,并进一步降解固体发酵疏松后的营养基质,进一步提升微生物对基质的利用效率,可最大化的提高枸杞提取物中纳豆激酶、蛋白酶和多肽等功能性成分的产量。
本发明将枸杞发酵液进一步与红枣粉末、木糖醇粉末混合生产出一种兼具多种营养价值的枸杞/红枣溶栓功能性饮品,该饮品中以木糖醇来提升饮品的甜度,木糖醇在体内的新陈代谢不需要胰岛素参与,又不会使血糖值升高,适合糖尿病类人群饮用;饮品中加入的红枣粉末不仅可以提升饮品的饮用价值,而且可以中和枸杞发酵液的口感;本发明制备的枸杞红枣复合发酵式功能饮品的色泽均匀、风味协调、清香甘醇、甜味适中、细腻无涩味、枸杞红枣香气甘醇,无异味。该产品中含有的酶和多肽,具有溶栓、抗氧化、抗衰老、美容养颜的功效;产品中含有枸杞多糖和蛋白质,具有补肾养肝、明目、润肺止咳、增强机体免疫力、抗肿瘤的功效,同时枸杞多糖甜度较低,可降血糖、降血脂,适宜作为糖尿病患者的食源之一。本发明公开的枸杞红枣复合发酵式功能饮品的原料组成虽然简单,但是却具有较高的营养价值和饮用价值。
附图说明
图1为实施例1、比较例1、比较例2中的三种不同发酵方法制备的枸杞发酵液在不同时间段的蛋白酶比活力;
图2为实施例1、比较例1、比较例2中的三种不同发酵方法制备的枸杞发酵液在不同时间段的纤溶活性;
图3为各实施例和比较例制备的枸杞红枣复合发酵式功能性饮品的感官评定结果。
具体实施方式
下面结合实施例对本发明进行详细说明。
本发明中所使用的纳豆枯草芽孢杆菌通过将购买于西安百川生物科技有限公司的纳豆粉经酪蛋白平板筛选得到;具体筛选方法为:将1mL不同浓度梯度的纳豆粉(1-10%)涂布于酪蛋白平板(脱脂奶粉5%,琼脂1.5%,其余为蒸馏水),37℃培养18-36h,每隔2h观察一次,最后选择透明圈最大的菌落。
实施例1
一种枸杞红枣复合发酵式功能性饮品的制备方法,包括以下步骤:
(1)将纳豆枯草芽孢杆菌于种子培养基中活化,种子培养基的配制为:胰蛋白胨6~8g、葡萄糖10~12g、牛肉浸膏3~5g、蒸馏水600mL、pH为7.2,再用酪蛋白平板筛选透明圈大的菌落,将较好菌落挑取至种子培养基中扩大培养,得到菌液浓度为1×107~1×108CFU/mL的菌液;
(2)将(1)中的菌液接种到由枸杞和无菌水按照1:1的质量比组成的枸杞发酵基质中,菌液的接种量为枸杞发酵基质质量的5%,然后在37℃,200r/min摇床培养6h进行固态发酵;
(3)将固态发酵结束后的基质转到锥形瓶中,加入62.5mL pH为6.8无菌水,在37℃,200r/min摇床培养6h进行液态发酵;
(4)将(3)中发酵6h后的液体发酵液过滤收集上清液,10000rpm下离心10min,得枸杞发酵液;
(5)将红枣干研磨成粉末;
(6)将木糖醇研磨成粉末;
(7)将枸杞发酵液30g、红枣粉末10g、木糖醇粉末5g混合,并加水至100mL,混合均匀,经过滤、杀菌、包装即得枸杞红枣复合发酵式功能性饮品。
实施例2
一种枸杞红枣复合发酵式功能性饮品的制备方法,其他同实施例1,只是在步骤(7)中,将枸杞发酵液40g、红枣粉末15g、木糖醇粉末7g混合,并加水至100mL,混合均匀,经过滤、杀菌、包装即得枸杞红枣复合发酵式功能性饮品。
实施例3
一种枸杞红枣复合发酵式功能性饮品的制备方法,其他同实施例1,只是在步骤(7)中,将枸杞发酵液50g、红枣粉末20g、木糖醇粉末10g混合,并加水至100mL,混合均匀,经过滤、杀菌、包装即得枸杞红枣复合发酵式功能性饮品。
比较例1
一种枸杞红枣复合发酵式功能性饮品的制备方法,包括以下步骤:
(1)将纳豆枯草芽孢杆菌于种子培养基中活化,种子培养基的配制为:胰蛋白胨6~8g、葡萄糖10~12g、牛肉浸膏3~5g、蒸馏水600mL、pH为7.2,再用酪蛋白平板筛选透明圈大的菌落,将较好菌落挑取至种子培养基中扩大培养,得到菌液浓度为1×107~1×108CFU/mL的菌液;
(2)将(1)中的菌液接种到由枸杞和无菌水按照1:1的质量比组成的枸杞发酵基质中,菌液的接种量为枸杞发酵基质质量的5%,然后在37℃,200r/min摇床培养12h进行固态发酵;
(3)将(2)中发酵后的固体培养基加入2倍质量的无菌生理盐水摇床37℃浸提30min后,浸提液过滤收集上清液为纳豆激酶粗酶液,过滤后的滤渣再用等体积的生理盐水于37℃条件下浸提30min,浸提液再次过滤,两次的浸提液在10000rpm下离心10min,得到枸杞发酵液。
(4)将红枣干研磨成粉末。
(5)将木糖醇研磨成粉末。
(6)将枸杞发酵液40g、红枣粉末15g、木糖醇粉末7g混合,并加水至100mL,混合均匀,经过滤、杀菌、包装,即得枸杞红枣复合发酵式功能性饮品。
比较例2
一种枸杞红枣复合发酵式功能性饮品的制备方法,包括以下步骤:
(1)将纳豆枯草芽孢杆菌于种子培养基中活化,种子培养基的配制为:胰蛋白胨6~8g、葡萄糖10~12g、牛肉浸膏3~5g、蒸馏水600mL、pH为7.2,再用酪蛋白平板筛选透明圈大的菌落,将较好菌落挑取至种子培养基中扩大培养,得到菌液浓度为0.71×108~0.82×108CFU/mL的菌液;
(2)将(1)中的菌液接种到由枸杞和无菌水按照1:1的质量比组成的枸杞发酵基质中,菌液的接种量为枸杞发酵基质质量的5%,然后加入62.5mL的无菌水,在37℃,200r/min摇床培养12h进行液态发酵;
(3)将(2)中发酵12h后的液体发酵液过滤收集上清液,10000rpm下离心10min,得到枸杞发酵液;
(4)将枸杞发酵液40g、红枣粉末16g、木糖醇粉末10g混合,并加水至100mL,混合均匀,经过滤、杀菌、包装,即得枸杞红枣复合发酵式功能性饮品。
实验例1
分别延长上述实施例1中的固态发酵时间为12h及液态发酵时间为12h、或固态发酵时间为24h及液态发酵时间为24h;比较例1中的固态发酵时间为24h、48h;比较例2中的液态发酵时间为24h、48h;分别应用福林酚法和纤维蛋白平板法测定枸杞发酵液中的蛋白酶比活力和纤溶活性,结果分别如图1、图2所示。由图可知实施例1中的固态发酵和液态发酵的时间共为12h时,蛋白酶活与纤维酶活达到最大,此时产品的蛋白酶降解和溶栓能力达到最大。同时,发酵时间成本低,产品的生产效率提升,成本降低。
测试例1
对上述实施例1、比较例1、比较例2中的枸杞发酵液中的蛋白质含量(凯氏定氮法)、多肽含量(双缩脲法)、抗氧化性(DPPH法和ABTS法)、总糖含量(GB/T 15672-2009)、还原糖含量(GB5009.7-85)、蛋白酶活力、纤溶酶活力进行测定,测定结果如表1所示。
表1
由表1可知,本发明公开的枸杞发酵液在固/液体顺序发酵与生物催化共反应条件下,各物质含量均高于独立固态发酵和液体发酵,突显出本专利的优势及商业价值。同时,该产品中含有的酶和多肽,具有溶栓、抗氧化、抗衰老、美容养颜的功效。产品中含有枸杞多糖和蛋白质,具有补肾养肝、明目、润肺止咳、增强机体免疫力、抗肿瘤的功效,同时枸杞多糖甜度较低,可降血糖、降血脂,适宜作为糖尿病患者的食源之一。
测试例2
枸杞红枣复合发酵式功能饮品按表2以色泽、风味、口感、香气为评定因素进行感官评价,评定小组由10位评价员组成,其中男生5名、女生5名,且在评定前一天不饮酒、抽烟,禁吃刺激性(过酸、过辣)食品,每评定一个样品结束后评定员要使用清水漱口,中间间隔10min后再对下一个样品进行评定,以保证评价结果真实、客观。感官评定结果见图3。
表2感官评定标准
由图3结果可知,实施例1-3中的枸杞红枣复合发酵式功能饮品在感官评分的分数上接近,相比于比较例1、比较例2而言有更优的感官评价。
上述参照实施例对枸杞红枣复合发酵式功能饮品及其制备方法进行的详细描述,是说明性的而不是限定性的,可按照所限定范围列举出若干个实施例,因此在不脱离本发明总体构思下的变化和修改,应属本发明的保护范围之内。
Claims (10)
1.一种枸杞红枣复合发酵式功能饮品的制备方法,其特征在于,包括以下步骤:
(1)将枸杞与无菌水等质量混合得到枸杞发酵基质,向枸杞发酵基质中接种纳豆芽孢杆菌的菌液,进行固态发酵;
(2)向固态发酵结束后的基质中加入无菌水,进行液态发酵;
(3)将液态发酵后的发酵液过滤,收集滤液,滤液离心后得到枸杞提取液;
(4)将枸杞提取液与红枣粉末、木糖醇粉末、水混合均匀,经过滤、杀菌、包装,即可得到所述枸杞红枣复合发酵式功能饮品。
2.根据权利要求1所述的制备方法,其特征在于,步骤(4)中,所述枸杞红枣复合发酵式功能饮品中各原料的质量百分比分别为:枸杞提取液30%~50%、红枣粉末10%~20%、木糖醇粉末5%~10%,其余为水。
3.根据权利要求1所述的制备方法,其特征在于,步骤(1)中,所述固态发酵的条件为35-40℃,150-220r/min摇床培养4-12h。
4.根据权利要求1所述的制备方法,其特征在于,步骤(2)中,所述液态发酵的条件为35-40℃,150-220r/min摇床培养4-12h。
5.根据权利要求1所述的制备方法,其特征在于,所述纳豆芽孢杆菌的菌液浓度为1×107~1×108CFU/mL,菌液的接种量为枸杞发酵基质质量的5%。
6.根据权利要求1或5所述的制备方法,其特征在于,所述纳豆芽孢杆菌的菌液的制备方法为:将纳豆芽孢杆菌于种子培养基中活化,再用酪蛋白平板筛选透明圈大的菌落,挑取筛选出的菌落至种子培养基中扩大培养。
7.根据权利要求6所述的制备方法,其特征在于,所述种子培养基的组成为:胰蛋白胨6~8g、葡萄糖10~12g、牛肉浸膏3~5g、蒸馏水600mL、pH为7.2。
8.根据权利要求1所述的制备方法,其特征在于,步骤(2)中的无菌水的加入量为枸杞质量的2.5~4倍。
9.根据权利要求1所述的制备方法,其特征在于,所述纳豆芽孢杆菌为纳豆枯草芽孢杆菌。
10.一种枸杞红枣复合发酵式功能饮品,其特征在于,采用权利要求1-9任意一项所述的制备方法制备得到。
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