CN111329055B - 一种簕菜酵素的制备方法 - Google Patents
一种簕菜酵素的制备方法 Download PDFInfo
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- CN111329055B CN111329055B CN202010159930.7A CN202010159930A CN111329055B CN 111329055 B CN111329055 B CN 111329055B CN 202010159930 A CN202010159930 A CN 202010159930A CN 111329055 B CN111329055 B CN 111329055B
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- acanthopanax
- trifoliate
- trifoliatus
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Abstract
本发明属于植物酵素加工技术领域,具体涉及一种簕菜酵素的制备方法。该制备方法是将簕菜原料打成浆,接着进行簕菜叶绿素提取,最后将提取后的滤渣与簕菜叶绿素提取液放入接种有益生菌和益生元的发酵罐中发酵,即得。本发明提供的簕菜酵素的制备方法先进行簕菜叶绿素提取,可以大大提高簕菜叶绿素含量高,保证簕菜酵素的功效发挥;同时其在发酵过程加入益生菌和益生元,可以更加充分的促进簕菜代谢产生大量的初级、次级代谢产物,提高簕菜酵素的营养成分,是一种较为理想的植物酵素饮料。
Description
技术领域
本发明属于植物酵素加工技术领域,具体涉及一种簕菜酵素的制备方法。
背景技术
酵素是一种由氨基酸组成的具有特殊生物活性的物质,存在于所有活的动植物体内,维持着机体的正常功能,参与机体的所有生命活动过程。植物酵素含有大量的β-胡萝卜素、维生素、蛋白质分解酵素、淀粉分解酵素、脂肪分解酵素、以及丰富的蛋白质、粗纤维、糖质、铁、钙、钠等成分的物质,其可以帮助人体改善肠胃生态环境,促进细胞营养吸收,激活细胞活力,中和体内过多的自由基,提高人体免疫力,受到了消费者极大的推崇。因此,越来越多的植物酵素应运而生。
专利文献CN105146614A公开了一种功能性钙果酵素、酵素饮料及其生产方法,该钙果酵素由钙果、枸杞、叶黄素、牛磺酸、蜂蜜、果葡糖浆、甜菊糖、低聚糖组成,其制备方法为:清洗、打浆、酶解、灭菌、发酵制得钙果酵素原液,由此经调配而成钙果酵素饮料。制得的钙果酵素及其饮料含有丰富的钙、多酚、黄酮、玉米黄质、β-胡萝卜素、多糖等特殊成分,对补钙、软化血管、改善微循环,提高机体抗氧化能力,延缓衰老、缓解视疲劳具有积极作用。
专利文献CN107811293A公开了一种辣木酵素,其是将辣木叶粉、大豆按一定比例混和,接种益生菌进行发酵,益生菌可以是米根霉、少孢根霉、纳豆芽孢杆菌,以辣木酵素为原料,可以生产出一系列营养、健康食品(饮料)。
专利文献CN109907307A公开了一种桑葚酵素,所述桑葚酵素由桑葚汁、薏仁粉、莲子粉、黄豆、乳酸菌、赤砂糖和水组成,其生产工艺包括如下步骤:1)黄豆的清洗、浸泡、水煮与过滤;2)第二、第三次水煮,滤液合并;3)将赤砂糖加入滤液,混匀,保温处理;4)加入薏仁粉、莲子粉,混合搅拌均匀;5)加入桑葚汁混合均匀;6)添加乳酸菌、水进行第一次发酵;7)排气,进行第二次发酵;8)升温、再次排气;9)降温保存,即得。制得的桑葚酵素具有抗氧化、增强机体免疫力,预防心脑血管疾病等作用。
据《中国高等植物图鉴》,簕菜,学名为白簕(Acanthopanan trifoliatus(L.)Merr.),又称鹅掌簕、三叶五加、三加皮、苦刺、刺三加,为五加科(Araliaceae)五加属(Acanthopanax)植物。广泛分布于我国中部和南部,印度、菲律宾、缅甸、泰国、越南和孟加拉国等地也有分布。簕菜是药食两用植物,簕菜产地民间自古就有采集簕菜作为鲜食蔬菜和保健药材的习俗。李时珍曰:“五加治风湿痪库,壮筋骨,其功良深”。明代《食物本草》中记有“五加,叶作蔬食,去皮肤风湿”。据《广东药用植物简编》(吴修仁编)记载,簕菜根、茎、叶均可入药,其辛、微苦、涩、凉,气微香。根,祛风除湿、散痕、止痛。叶,疏风,消肿,止痒,有舒筋活血、消肿解毒之功效。据《中国高等植物图鉴》,簕菜味苦、辛、凉,具有清热解毒、祛风利湿、舒筋活血、止咳平喘之功效。簕菜在广东江门市的恩平等地盛产,并且十多年前已经开始了人工种植。目前为止,并未见有关于簕菜发酵方面的研究报道。
发明内容
为了拓宽簕菜的应用价值,本发明提供了一种簕菜酵素的制备方法,制备得到的簕菜酵素叶绿素含量高,稳定性好,是一种较为理想的植物酵素饮料。
本发明提供的簕菜酵素的制备方法,包括以下步骤:
S1.称取簕菜,清洗后离心去水分,加入无菌水后倒入磨浆机中磨浆,所述无菌水的添加量是簕菜总重量的8~12%,过80~120目,得簕菜浆液;
S2.往步骤S1得到的簕菜浆液中加入乙醇溶液,所述乙醇溶液的添加量为簕菜浆液总体积的6~8倍,高压脉冲电场处理8~10h,过滤,滤渣备用,将滤液进行真空精馏,回收乙醇,得簕菜叶绿素提取液;
S3.将步骤S2得到的簕菜叶绿素提取液和滤渣通过无菌管道输送到发酵罐,同时接入混合菌种,所述混合菌种由卷曲乳杆菌、德氏乳杆菌乳亚种、格氏乳杆菌、约氏乳杆菌和唾液乳杆菌按1:1:1:1:1的比例混合组成,接入的混合菌种的总体积是簕菜浆液总体积的1/1000;
S4.将益生元加入步骤S3的发酵罐中,搅拌均匀,发酵4~5周,所述发酵条件为:混合菌种接种后第一周,温度设置为33~35℃,混合菌种接种后第二周开始温度设置为28-32℃,得发酵液;
S5.将步骤S4得到的发酵液进行负高压脉冲处理3~5min,装瓶,即得。
进一步地,所述步骤S2中的乙醇溶液为体积浓度为70~80%的乙醇溶液。
进一步地,所述步骤S2中高压脉冲电场的条件为:电场场强为30~40kV/cm,频率为4~6Hz。
进一步地,所述步骤S2中高压脉冲电场处理的条件为:电场场强为36kV/cm,频率为5Hz。
进一步地,所述步骤S3中混合菌种的活菌数为5*109~7*109cfu/mL。
进一步地,所述步骤S4中的益生元由低聚果糖、低聚半乳糖、低聚木糖和菊粉按质量比(8~10):1:1:(4~6)组成。
进一步地,所述步骤S4中的益生元由低聚果糖、低聚半乳糖、低聚木糖和菊粉按质量比9:1:1:5组成。
进一步地,所述步骤S4中益生元的添加量为簕菜浆液总体积的15~20%。
进一步地,所述步骤S5中负高压脉冲处理的条件为:负脉冲高压为-10~-5kV,频率为2~4Hz。
另外,本发明还提供了所述的簕菜酵素制备方法制得的簕菜酵素在制备防治便秘的食品或保健品中的用途。
本发明提供的益生菌为:卷曲乳杆菌(Lactobacillus crispatus)、德氏乳杆菌乳亚种(Lactobacillus delbrueckii subsp.Lactis)、格氏乳杆菌(Lactobacillusgasseri)、约氏乳杆菌(Lactobacillus johnsonii)、唾液乳杆菌(Lactobacillussalivarius)。本发明制得的簕菜酵素饮料干燥后簕菜酵素可做成簕菜酵素固体饮料或压片糖果。
目前,现有技术制备植物酵素的常规方法是采用益生菌进行发酵,用于分解出植物原料的营养物质,然而单单采用益生菌进行发酵容易出现菌种活力低、发酵不充分的现象,导致植物酵素的有机活性成分含量较低,大大降低植物酵素的效果。
为了解决上述问题,本发明人经过大量的摸索实验发现:在发酵过程中引入由低聚果糖、低聚半乳糖、低聚木糖和菊粉按特定质量比组成的益生元,不仅可以提高益生菌在发酵过程中的活力,使簕菜代谢产生大量的初级、次级代谢产物如氨基酸、维生素、有机酸、各种酶、芳香族化合物、糖醇、多糖、小分子肽等有机活性成分,从而提高簕菜的发酵效果。同时,该特定成分的益生元还可以用于调节人体肠胃的益生菌,促进人体肠道有益菌群的增值,抑制有害菌,从而减少腐败物质和有毒物质的产生,进一步地提高簕菜酵素增强免疫、降血压血脂,平衡血糖,增强肝脏代谢等功能。
另外,虽然簕菜的叶绿素含量高,但是由于叶绿素本身溶出度低,稳定差,叶绿素遇光、酸、碱、氧、氧化剂等都会使其分解,尤其在酸性条件下,叶绿素分子很容易失去卟啉环中的镁成为去镁叶绿素等性质,导致簕菜叶绿素在人体内无法有效发挥作用。
而本发明提供的簕菜酵素的制备方法可以有效的解决簕菜叶绿素在人体内的稳定性,保证簕菜酵素的功效发挥。本发明在对簕菜进行叶绿素提取时,采用电场场强为30~40kV/cm,频率为4~6Hz的特定高压脉冲电场处理,可以大大提高簕菜叶绿素的提取率。其具体的原理为:本发明提供的特定高压脉冲电场处理可以加快簕菜浆液内分子的运动,提高叶绿素的溶出度,同时其还可以降低簕菜浆液的颗粒度,增大颗粒与乙醇溶度的接触面积,从而提高绿叶素的溶出度。然而采用高于或低于本发明提供的电场场强处理和脉冲均不能提高叶绿素的溶出度。
进一步地,本发明在发酵后将发酵液再经过负脉冲高压为-10~-5kV,频率为2~4Hz的特定负高压脉冲处理,可以大大提高绿叶素的稳定性。其具体的原理为:本发明提供的特定负高压脉冲处理可以使发酵液能够迅速的离解出带电离子,而且该负脉冲电场使得发酵液中的正电荷朝向内部流动,而负电荷将聚集于发酵液的表面,这样不仅可以防止发酵液中叶绿素分子失去卟啉环中的镁成为去镁叶绿素,提高叶绿素的稳定性,同时,溶液表面聚集大量负电荷离子的发酵液可以中和胃酸中游离酸的氢离子,从而减少胃酸对叶绿素的破坏,保证簕菜叶绿素在人体内的稳定性。然而采用高于或低于本发明提供的负脉冲高压和频率均不能提高叶绿素的稳定性。
总之,与现有技术,本发明提供簕菜酵素的制备方法具有有机活性成分高、营养丰富,叶绿素含量高和稳定性好的优点,是一种较为理想的植物酵素饮料。
具体实施方式
以下通过具体实施例进一步描述本发明,本发明不仅仅限于以下实施例。在本发明的范围内或者在不脱离本发明的内容、精神和范围内,对本发明进行的变更、组合或替换,对于本领域的技术人员来说是显而易见的,且包含在本发明的范围之内。
实施例1、簕菜酵素的制备
S1.称取簕菜,清洗后离心去水分,加入无菌水后倒入磨浆机中磨浆,所述无菌水的添加量是簕菜总重量的8%,过80目,得簕菜浆液;
S2.往步骤S1得到的簕菜浆液中加入体积浓度为70%的乙醇溶液,所述乙醇溶液的添加量为簕菜浆液总体积的6倍,高压脉冲电场处理8h,所述高压脉冲电场的条件为:电场场强为30kV/cm,频率为6Hz,过滤,滤渣备用,将滤液进行真空精馏,回收乙醇,得簕菜叶绿素提取液;
S3.将步骤S2得到的簕菜叶绿素提取液和滤渣通过无菌管道输送到发酵罐,同时接入混合菌种,所述混合菌种由卷曲乳杆菌、德氏乳杆菌乳亚种、格氏乳杆菌、约氏乳杆菌和唾液乳杆菌按1:1:1:1:1的比例混合组成,所述混合菌种的活菌数为5*109cfu/mL,接入的混合菌种的总体积是簕菜浆液总体积的1/1000;
S4.将益生元加入步骤S3的发酵罐中,所述益生元由低聚果糖、低聚半乳糖、低聚木糖和菊粉按质量比8:1:1:6组成,所述益生元的添加量为簕菜浆液总体积的15%,搅拌均匀,发酵4周,所述发酵条件为:混合菌种接种后第一周,温度设置为33~35℃,混合菌种接种后第二周开始温度设置为28-32℃,得发酵液;
S5.将步骤S4得到的发酵液进行负高压脉冲处理3min,所述负高压脉冲处理的条件为:负脉冲高压为-10kV,频率为2Hz,装瓶,即得。
实施例2、簕菜酵素的制备
S1.称取簕菜,清洗后离心去水分,加入无菌水后倒入磨浆机中磨浆,所述无菌水的添加量是簕菜总重量的10%,过100目,得簕菜浆液;
S2.往步骤S1得到的簕菜浆液中加入体积浓度为75%的乙醇溶液,所述乙醇溶液的添加量为簕菜浆液总体积的7倍,高压脉冲电场处理9h,所述高压脉冲电场的条件为:电场场强为36kV/cm,频率为5Hz,过滤,滤渣备用,将滤液进行真空精馏,回收乙醇,得簕菜叶绿素提取液;
S3.将步骤S2得到的簕菜叶绿素提取液和滤渣通过无菌管道输送到发酵罐,同时接入混合菌种,所述混合菌种由卷曲乳杆菌、德氏乳杆菌乳亚种、格氏乳杆菌、约氏乳杆菌和唾液乳杆菌按1:1:1:1:1的比例混合组成,所述混合菌种的活菌数为6*109cfu/mL,接入的混合菌种的总体积是簕菜浆液总体积的1/1000;
S4.将益生元加入步骤S3的发酵罐中,所述益生元由低聚果糖、低聚半乳糖、低聚木糖和菊粉按质量比9:1:1:5组成,所述益生元的添加量为簕菜浆液总体积的18%,搅拌均匀,发酵4周,所述发酵条件为:混合菌种接种后第一周,温度设置为33~35℃,混合菌种接种后第二周开始温度设置为28-32℃,得发酵液;
S5.将步骤S4得到的发酵液进行负高压脉冲处理4min,所述负高压脉冲处理的条件为:负脉冲高压为-8kV,频率为3Hz,装瓶,即得。
实施例3、簕菜酵素的制备
S1.称取簕菜,清洗后离心去水分,加入无菌水后倒入磨浆机中磨浆,所述无菌水的添加量是簕菜总重量的12%,过120目,得簕菜浆液;
S2.往步骤S1得到的簕菜浆液中加入体积浓度为80%的乙醇溶液,所述乙醇溶液的添加量为簕菜浆液总体积的8倍,高压脉冲电场处理10h,所述高压脉冲电场的条件为:电场场强为40kV/cm,频率为4Hz,过滤,滤渣备用,将滤液进行真空精馏,回收乙醇,得簕菜叶绿素提取液;
S3.将步骤S2得到的簕菜叶绿素提取液和滤渣通过无菌管道输送到发酵罐,同时接入混合菌种,所述混合菌种由卷曲乳杆菌、德氏乳杆菌乳亚种、格氏乳杆菌、约氏乳杆菌和唾液乳杆菌按1:1:1:1:1的比例混合组成,所述混合菌种的活菌数为7*109cfu/mL,接入的混合菌种的总体积是簕菜浆液总体积的1/1000;
S4.将益生元加入步骤S3的发酵罐中,所述益生元由低聚果糖、低聚半乳糖、低聚木糖和菊粉按质量比10:1:1:4组成,所述益生元的添加量为簕菜浆液总体积的20%,搅拌均匀,发酵4~5周,所述发酵条件为:混合菌种接种后第一周,温度设置为33~35℃,混合菌种接种后第二周开始温度设置为28-32℃,得发酵液;
S5.将步骤S4得到的发酵液进行负高压脉冲处理3~5min,所述负高压脉冲处理的条件为:负脉冲高压为-5kV,频率为4Hz,装瓶,即得。
对比例1、簕菜酵素的制备
与实施例2的区别在于:所述步骤S2不进行高压脉冲电场处理,其余步骤与实施例2类似。
对比例2、簕菜酵素的制备
与实施例2的区别在于:所述步骤S2采用超声震动提取替代高压脉冲电场处理,其余步骤与实施例2类似。
对比例3、簕菜酵素的制备
与实施例2的区别在于:所述步骤S4不添加益生元直接进行发酵处理,其余步骤与实施例2类似。
对比例4、簕菜酵素的制备
与实施例2的区别在于:所述步骤S5不进行负高压脉冲处理处理,其余步骤与实施例2类似。
对比例5、簕菜酵素的制备
与实施例2的区别在于:所述步骤S5进行高压脉冲处理处理,所述高压脉冲处理的条件为:电场场强为36kV/cm,频率为5Hz,其余步骤与实施例2类似。
试验例一、簕菜酵素的叶绿素含量检测试验
1、试验对象:
实施例1、实施例2、实施例3、对比例1和对比例2步骤S2制备得到的簕菜叶绿素提取液。
2、试验方法:
取1kg簕菜,按照实施例1、实施例2、实施例3、对比例1和对比例2的步骤S1和步骤S2提取叶绿素,将提取得到的簕菜叶绿素提取液在50℃下烘干,测定簕菜叶绿素的含量。
3、试验结果
试验结果如表1所示。
表1簕菜酵素的叶绿素含量测定数据
实施例1 | 实施例2 | 实施例3 | 对比例1 | 对比例2 | |
叶绿素含量(g) | 1.02 | 1.08 | 1.00 | 0.65 | 0.73 |
由表1可知,本发明提供的簕菜酵素的制备方法可以有效的提高簕菜叶绿素的含量,保证簕菜酵素的功效发挥,提高簕菜酵素的产品质量。
试验例二、簕菜酵素的润肠通便效果
1、试验对象:
选取健康Balb/c小鼠70只,购自广东省医学实验动物中心,雄性,体重18~22g。
2、试验材料:
实施例2、对比例3、对比例4和对比例5制得的簕菜酵素,酚酞含片,购于广东台城制药股份有限公司(国药准字H44024959),复方地芬诺酯,购于长春长红制药有限公司(国药准字H22022037)。
3、试验方法:
将小鼠随机分为7组,每组10只。分别为正常对照组、模型组、阳性对照组、实施例2组、对比例3组、对比例4组和对比例5组。给各组小鼠进行灌胃给药,持续15天,剂量分别如下:
正常对照组:给予相同体积的蒸馏水灌胃;
模型组:给予相同体积的蒸馏水灌胃;
阳性对照组:给予1g/kg酚酞含片灌胃;
实施例2组:给予4g/kg的实施例2制备的簕菜酵素灌胃;
对比例3组:给予4g/kg的对比例3制备的簕菜酵素灌胃;
对比例4组:给予4g/kg的对比例4制备的簕菜酵素灌胃;
对比例5组:给予4g/kg的对比例5制备的簕菜酵素灌胃;
给药结束后,各组小鼠禁食不禁水16h,模型组、阳性对照组、实施例2组、对比例3组、对比例4组和对比例5组通过灌胃复方地芬诺酯,构建便秘动物模型,正常对照组灌胃生理盐水。灌胃复方地芬诺酯0.5h后,阳性对照组灌胃含酚酞含片的墨汁,实施例2组灌胃实施例2制备的簕菜酵素墨汁,对比例3组灌胃对比例3制备的簕菜酵素墨汁,对比例4组灌胃对比例4制备的簕菜酵素墨汁,对比例5组灌胃对比例5制备的簕菜酵素墨汁,正常对照组和模型组灌胃墨汁。从灌胃墨汁开始,记录每只小鼠排出首粒黑便的时间和8h内和粪便重量。
4、试验结果:
试验结果如表2所示。
表2簕菜酵素对小鼠的排便影响
与正常对照组相比,模型组#P<0.05,##P<0.01。与模型组相比,*P<0.05,**P<0.01。
由表2可知,本发明制备得到的簕菜酵素能显著改善模型小鼠的润肠通便能力,与模型组相比,首次排黑便时间大大缩短,8h内排粪便重量显著增加。
Claims (7)
1.一种簕菜酵素的制备方法,其特征在于,包括以下步骤:
S1.称取簕菜,清洗后离心去水分,加入无菌水后倒入磨浆机中磨浆,所述无菌水的添加量是簕菜总重量的8~12%,过80~120目,得簕菜浆液;
S2.往步骤S1得到的簕菜浆液中加入乙醇溶液,所述乙醇溶液的添加量为簕菜浆液总体积的6~8倍,高压脉冲电场处理8~10h,过滤,滤渣备用,将滤液进行真空精馏,回收乙醇,得簕菜叶绿素提取液;
S3.将步骤S2得到的簕菜叶绿素提取液和滤渣通过无菌管道输送到发酵罐,同时接入混合菌种,所述混合菌种由卷曲乳杆菌、德氏乳杆菌乳亚种、格氏乳杆菌、约氏乳杆菌和唾液乳杆菌按1:1:1:1:1的比例混合组成,接入的混合菌种的总体积是簕菜浆液总体积的1/1000;
S4.将益生元加入步骤S3的发酵罐中,搅拌均匀,发酵4~5周,所述发酵条件为:混合菌种接种后第一周,温度设置为33~35℃,混合菌种接种后第二周开始温度设置为28-32℃,得发酵液;
S5.将步骤S4得到的发酵液进行负高压脉冲处理3~5min,装瓶,即得;
所述步骤S4中的益生元由低聚果糖、低聚半乳糖、低聚木糖和菊粉按质量比(8~10):1:1:(4~6)组成;
所述步骤S5中负高压脉冲处理的条件为:负脉冲高压为-10~-5kV,频率为2~4Hz;
所述步骤S2中高压脉冲电场的条件为:电场场强为30~40kV/cm,频率为4~6Hz。
2.如权利要求1所述的簕菜酵素的制备方法,其特征在于,所述步骤S2中的乙醇溶液为体积浓度为70~80%的乙醇溶液。
3.如权利要求1所述的簕菜酵素的制备方法,其特征在于,所述步骤S2中高压脉冲电场处理的条件为:电场场强为36kV/cm,频率为5Hz。
4.如权利要求1所述的簕菜酵素的制备方法,其特征在于,所述步骤S3中混合菌种的活菌数为5*109~7*109cfu/mL。
5.如权利要求1所述的簕菜酵素的制备方法,其特征在于,所述步骤S4中的益生元由低聚果糖、低聚半乳糖、低聚木糖和菊粉按质量比9:1:1:5组成。
6.如权利要求1所述的簕菜酵素的制备方法,其特征在于,所述步骤S4中益生元的添加量为簕菜浆液总体积的15~20%。
7.如权利要求1~6任一所述的簕菜酵素制备方法制得的簕菜酵素在制备预防便秘的保健品中的用途。
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