CN115590894A - 一种海藻全活性物质的制备方法 - Google Patents
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Abstract
本发明公开了一种海藻全活性物质的制备方法,所述海藻全活性物质利用超声同时酶解的方法制备。本发明提供了一种海藻活性物质的零浪费制备方法,该制备方法可以提高产率、减少污染、降低生产成本,并具有一定普适性,有着广阔的应用前景。
Description
技术领域
本发明属于海藻全活性物质超声同时酶解生产工艺,涉及一种海藻活性物质的零浪费制备方法,该活性物质具有很高的营养价值及药用价值,制备的海藻活性物质具备较高的生物活性,促进植物根的生长、抗菌,提高人体免疫力、抗肿瘤、增强体力等。
背景技术
海藻是指生长在潮间带及亚潮间带肉眼可见的大型藻类,通常包括绿藻、褐藻及红藻三大类。在古代中国及日本就有利用海藻做为食物的证据,古医典包括《本草纲目》、《本草经集注》、《海药本草》及《本草拾遗》等都有用海藻治疗各种疾病的纪载。许多种海藻依其化学成分不同而有不同的药理作用及临床用途。可以用于退烧、治咳,以及治疗气喘、痔疮、流鼻涕、肠胃不适及泌尿疾病等。还可以加强身体抗癌、抗肿瘤的能力,且可有效改善糖尿病症状及纾解紧张压力。
海藻中的活性成分丰富,包括多糖、多酚等丰富的活性物质,但目前工业上对海藻活性物质的提取方法单一,运用大量的有机试剂等不仅容易导致产品降解严重、黏度降低,还会对环境造成严重的污染。近年来,又逐渐发展了以酶解或超声处理代替有机体系浸泡的酶解法和超声法。酶解法是在一定条件下用酶分解海藻细胞壁,促进海藻活性物质溶出,从而提高海藻活性物质产率的方法。酶解法具有专一性强、条件温和、工艺简单等优点。但是,单一酶存在酶解效果不佳、生产成本高、工艺周期长且产品纯度不高等问题,超声法利用超声波使海藻细胞壁在瞬间破裂,并通过振动作用和热效应加速细胞内有效物质的释放、扩散和溶解,有助于海藻活性物质溶出,使产率显著提高,但是超声法耗时较长,目前超声和酶解也多为单独使用,并未得到有效结合,工艺方法尚未成熟。
超声-复合分步酶解法能有效破坏海藻细胞壁的结构,不仅能够提高海藻活性物质的产率、缩短生产周期、减少环境污染,还通过减少酸和碱的用量,有效降低生产成本。同是该方法适用于所有大型海藻,对各种海藻多糖的提取有参考价值,对大型海藻类生物质资源的高附加值利用有借鉴意义。本发明中通过超声-复合分步酶解法制备具有抗氧化活性的海藻全活性物质均未见其它研究报道或专利。
发明内容
本发明的目的是提供一种海藻全活性物质的绿色制备方法。制备工艺路线简单合理、绿色高效,是一种制备海藻全活性物质的有效方法。为实现上述目的,本发明采用如下技术方案:
一种海藻全活性物质的制备方法,包含有如下步骤:
(1)将海藻原料洗净后烘干,之后用粉碎机粉碎成海藻颗粒(或粗粉); (2)往海藻粉中加入10-20倍重量的体积分数55%-75%乙醇溶液为提取溶剂在65-80℃回流提取2-3h,用50-500目尼龙布过滤获得海藻醇提液和海藻渣1;
(3)往海藻渣1中加入10-50倍重量的蒸馏水,加入海藻渣1重量0.1-0.5%纤维素酶+0.1-0.5%果胶酶,调整pH=3-6,酶解和超声处理2-3h后,加入海藻渣1重量0.1-0.5%复合蛋白酶+0.2-0.6%木瓜蛋白酶,调整pH=6-9, 酶解和超声处理2-3h后,用90-130℃灭酶10-20min后,过滤,得到海藻活性物质提取液和海藻渣2回收烘干。
(4)往海藻渣2中加入其质量1-5%的无水碳酸钠以及20-50倍重量的蒸馏水,55-85℃超声提取2-3h,离心过滤得滤渣放入烘箱进行烘干得海藻不可溶性膳食纤维,滤液浓缩后,热风干燥、粉碎获得可溶性膳食纤维。
进一步的,步骤(1)中,所述海藻可以是绿藻、红藻、褐藻等,应用范围广。
进一步的,步骤(2)(3)(4)中,所述的加工工艺中所用试剂都是无毒安全试剂。
进一步的,步骤(3)中,超声-复合分步酶解法能有效破坏海藻细胞壁的结构,不仅能够提高海藻活性物质的产率、缩短生产周期、减少环境污染,还通过减少酸和碱的用量,有效降低生产成本。
进一步的,步骤(2)(3)(4)中,所述的提取过程将海藻的所有活性物质进行提取,做到零浪费,操作简便,同时可以连续地大规模进行生产,适合大规模化工业生产。
本发明的优点是:
(1)本发明的提取方法合理有效,利用超声-复合分步酶解的零浪费制备方法制备,具有无副反应、条件温和、环境友好、节约能耗等优点;
(2)本发明采购所有海藻进行提取,极大的利用了海藻资源;
(3)本发明所有超声-复合分步酶解法能有效破坏海藻细胞壁的结构,不仅能够提高海藻活性物质的产率、缩短生产周期、减少环境污染,还通过减少酸和碱的用量,有效降低生产成本。
具体实施方式
下面通过实施例对本发明做进一步详细描述, 但本发明不仅仅局限于以下实施例。
实施例1
一种海藻全活性物质的绿色制备方法,包括以下步骤:
(1)将海带原料洗净后烘干,之后用粉碎机粉碎成海带颗粒(或粗粉); (2)往海带粉中加入20倍重量的体积分数55%乙醇溶液为提取溶剂在65℃回流提取2h,用300目尼龙布过滤获得海带醇提液和海带渣1;
(3)往海带渣1中加入40倍重量的蒸馏水,加入海带渣1重量0.1%纤维素酶+0.1%果胶酶,调整pH=3,酶解和超声处理2h后,加入海带渣1重量0.1%复合蛋白酶+0.2%木瓜蛋白酶,调整pH=6, 酶解和超声处理2h后,用100℃灭酶10min后,过滤,得到海带活性物质提取液和海带渣2回收烘干。
(4)往海带渣2中加入其重量1%的无水碳酸钠以及30倍重量的蒸馏水,55℃超声提取2h。离心过滤得滤渣放入烘箱进行烘干得海带不可溶性膳食纤维,滤液浓缩后,热风干燥、粉碎获得可溶性膳食纤维。
(5)对海带全活性物质得率及其活性物质含量进行测定。结果如下:
1. 海带全活性物质各物质产量及得率
2.多糖测定
多糖含量的具体测定方法为吸取0、0.2、0.4、0.6、0.8、1.0 mL的0.1mg/mL的标准葡萄糖溶液于试管中,蒸馏水补至1.0 mL,加入5%的苯酚溶液1.0 mL和5.0 mL的硫酸溶液,避光静置10 min,涡旋混匀,转入30 ℃水浴锅中反应20 min,吸取200 μL于96孔板中490nm处测吸光值。得出葡萄糖标准曲线为:y = 0.00061x + 0.1919,R² = 0.9983。
3. 蛋白测定
采用BCA试剂盒法测定海带活性物质中的蛋白含量。按照试剂盒的说明书配置取的蛋白标准溶液(20 mg/mL),将标准溶液依次0、1、2、4、8、12、16、20μL的加到96孔板中,加稀释液补足至20μL,其蛋白浓度依次为0、25、50、100、200、300、400、500μg/mL,加20μL的待测样品到96孔板中。37℃孵育20-30min,在酶标仪562nm处测定吸光度,根据标准曲线计算出待测样品的蛋白浓度。蛋白标准曲线为:y = 0.0009x + 0.0827,R² = 0.9971。
本发明能够提高海藻活性物质的产率,缩短生产周期。
以上所述仅为本发明的较佳实施例,凡依本发明申请专利范围所做的均等变化与修饰,皆应属本发明的涵盖范围。
Claims (4)
1.一种海藻全活性物质的制备方法,其特征在于,包含有如下步骤:
(1)将海藻原料洗净后烘干、粉碎,加入10-20倍重量的体积分数55%-75%乙醇溶液在65-80℃回流提取2-3h,用50-500目尼龙布过滤获得海藻醇提液和海藻渣1;
(2)往海藻渣1中加入10-50倍重量的蒸馏水,加入0.1-0.5wt%纤维素酶和0.1-0.5wt%果胶酶 ,调整pH=3-6,酶解和超声处理2-3h后,加0.1-0.5wt%复合蛋白酶和0.2-0.6wt%木瓜蛋白酶,调整pH=6-9,酶解和超声处理2-3h后,用90-130℃灭酶10-20min后,过滤,得到海藻活性物质提取液和海藻渣2回收烘干;
(3)往海藻渣2中加入其质量1-5%的无水碳酸钠以及20-50倍重量的蒸馏水,55-85℃超声提取2-3h,离心过滤得滤渣放入烘箱进行烘干得海藻不可溶性膳食纤维,滤液浓缩后,热风干燥、粉碎获得可溶性膳食纤维。
2. 根据权利要求 1 所述的制备方法,其特征在于,所述海藻包括绿藻、红藻、褐藻。
3. 根据权利要求 1 所述的制备方法,其特征在于,所述海藻为海带。
4. 根据权利要求 1 所述的制备方法,其特征在于,所述海藻全活性物质包括海藻醇提液、海藻活性物质提取液、不可溶性膳食纤维和可溶性膳食纤维。
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