CN111109604A - 一种具有抗氧化活性的枸杞发酵物及其制备方法 - Google Patents
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Abstract
本发明涉及微生物技术领域,公开了一种具有抗氧化活性的枸杞发酵物,该枸杞发酵物通过冠突散囊菌发酵枸杞制备而成,其制备方法包括原料准备、打浆、高压灭菌、接种冠突散囊菌、发酵、过滤等步骤,本发明采用冠突散囊菌发酵枸杞得到枸杞发酵物,发酵工艺简单、周期短、操作温度低,枸杞发酵物清除DPPH、ABTS自由基的活性均优于未发酵的枸杞液,具有显著的抗氧化活性,该发明为进一步合理利用枸杞资源奠定了基础,提高了枸杞的农业附加值,适用于食品、保健品、化妆品或药品等领域。
Description
技术领域
本发明涉及微生物技术领域,具体的说涉及一种具有抗氧化活性的枸杞发酵物及其制备方法。
背景技术
枸杞,属茄科落叶灌木,我国为枸杞主产地,尤以宁夏为主产区。枸杞在我国已有2330多年的历史,枸杞乃是我国传统的名贵中药材,《本草纲目》记载枸杞具有滋补身体、益精明目、增强体质等作用;同时枸杞也可泡茶、泡酒、熬粥以及直接食用,是一种药食两用的食材。枸杞具有多糖、多酚、黄酮等多种活性成分,具有抗氧化、改善糖尿病并发症、调节体质代谢等作用。
冠突散囊菌,俗称金花菌,是一种益生菌,主要用于茯砖茶的生茶发酵。冠突散囊菌在发酵过程中会产生蛋白酶、淀粉酶以及脂肪酶等酶,不仅可以改善发酵液的香味,还可以产生许多的活性物质,如胞外多糖具有维持血脂平衡、调节胆固醇代谢以及抑制氧化细胞生长等功效;多酚类化合物具有较强的抗氧化、抗衰老等功效。
目前,关于枸杞的研究越来越多,枸杞果酒、枸杞保健饮料、枸杞休闲食品相继开发,但是现有枸杞产品的多酚类物质含量较低,限制了其抗氧化活性,从而使得枸杞的农业附加值较低,影响了枸杞种植业的经济效益。
发明内容
针对现有技术中的不足,本发明要解决的技术问题在于提供了一种具有抗氧化活性的枸杞发酵物及其制备方法。
为解决上述技术问题,本发明通过以下方案来实现:一种具有抗氧化活性的枸杞发酵物,该枸杞发酵物通过冠突散囊菌发酵枸杞制备而成,其具有显著的抗氧化活性。
本发明的一种枸杞发酵物的制备方法,该制备方法包括如下步骤:
(1)原料准备:取适量湿枸杞清洗备用;
(2)打浆:按质量比为1:20加入蒸馏水,并用打浆器将枸杞的浆水打出,并搅拌均匀;
(3)高压灭菌:将步骤2中的均浆液体分装至三角瓶,每瓶200 ml,密封后进行高压灭菌,冷却备用;
(4)接种冠突散囊菌:将步骤3中的匀浆液体在无菌条件下按体积比1:400接种冠突散囊菌液;
(5)发酵:将步骤4中的液体密封、摇匀,置于震荡培养箱中进行发酵,发酵温度为28 °C,震荡培养箱的转速为200 r/min,发酵时间为1-15天;
(6)过滤:将步骤5中发酵后的发酵混合物过滤,得到液体枸杞发酵物。
进一步的,所述步骤4中冠突散囊菌液的菌种密度为1×10 6/ml。
一种枸杞发酵物在制备食品、保健品、化妆品或药品中的应用。
相对于现有技术,本发明的有益效果是:本发明首次将冠突散囊菌用于枸杞的发酵制备,发酵后可提高枸杞发酵物中活性成分——多酚类物质的含量,增强枸杞发酵物的抗氧化活性,从而提高了枸杞的农业附加值,提高了枸杞种植业的经济效益。本发明枸杞发酵物的制备方法工艺简单、生产周期短,同时制备的枸杞发酵物具有良好的抗氧化活性,保健效果好,在食品、保健品、化妆品或药品中的应用具有广阔的市场前景。
附图说明
图1为本发明枸杞发酵物的制备流程示意图;
图2为本发明不同发酵天数枸杞发酵物的DPPH自由基清除率对比示意图;
图3为本发明不同发酵天数枸杞发酵物的ABTS自由基清除率对比示意图。
具体实施方式
下面结合附图对本发明的优选实施例进行详细阐述,以使发明的优点和特征能更易于被本领域技术人员理解,从而对本发明的保护范围做出更为清楚明确的界定。
实施例1
本发明的一种具有抗氧化活性的枸杞发酵物,该枸杞发酵物通过冠突散囊菌发酵枸杞制备而成,该枸杞发酵物的多酚类活性成分较多,具有显著的抗氧化活性。
本优选实施例中,枸杞发酵物的制备方法包括如下步骤:
(1)原料准备:取70 g湿枸杞清洗备用;
(2)打浆:加入1400 ml蒸馏水,用打浆器将枸杞的浆水打出,并搅拌均匀;
(3)高压灭菌:将步骤2中的均浆液体分装至7个三角瓶中,每瓶200 ml,对每个三角瓶密封后进行高压灭菌,冷却备用;
(4)接种冠突散囊菌:将步骤3中每个三角瓶内的匀浆液体在无菌条件下接种0.5ml冠突散囊菌液,冠突散囊菌液的菌种密度为1×10 6/ml。
(5)发酵:将步骤4中每个三角瓶的液体进行密封、摇匀,置于震荡培养箱中进行发酵,发酵温度为28 °C,震荡培养箱的转速200 r/min,7个三角瓶的发酵时间分别为0、5、7、9、11、13、15天;
(6)过滤:将步骤5中发酵后的发酵混合物过滤,得到液体枸杞发酵物。
实施例2
本实施例对上述提取获得的枸杞发酵液中多酚含量进行了测量,其方法及测量结果如下:
Folin-Ciocalteu法:以没食子酸为标准物质测定多酚含量。取20 μL样品加入96孔板,加入Folin-Ciocalteu试剂100 μL、10%Na2CO3溶液25 μL,反应温度37 ℃,反应时间1.0 h,以吸光度为评价指标,检测波长765 nm。吸光度与多酚含量在0~250 μg/mL范围内呈良好的线性关系(y=0.0085x+0.0354,R2=0.9997)。
测量结果如表1所示,经冠突散囊菌发酵获取的枸杞发酵液多酚含量显著提高,高于未发酵的枸杞液;其中,发酵后第9天其多酚含量最高,为13.058 μg/mL,比未发酵的多酚含量提高了78.39%。
表1 不同发酵时间枸杞发酵提取物的多酚含量
发酵天数 | 多酚含量(μg/mL) |
0 | 7.32 |
5 | 11.33 |
7 | 11.21 |
9 | 13.06 |
11 | 11.75 |
13 | 11.91 |
15 | 10.26 |
实施例3
本实施例通过以下两种方法对上述提取获得的枸杞发酵液进行抗氧化活性评价:
(1)DPPH清除率测定:取乙醇溶解的DPPH(0.05 mg/mL)溶液2 mL加入到小试管中,加95%乙醇(或无水乙醇) 1 mL,充分混合,在517 nm波长处测定其吸光度ADPPH。依次分别加入100 μL不同发酵天数的枸杞发酵液,混匀,静置反应30 min后在517 nm波长处测定其吸光度A,以Vc为阳性对照。按照公式(1)计算不同天数发酵物DPPH自由基清除率:
式中:ADPPH为对照组DPPH溶液的吸光度;A样品为加入样品溶液或Vc后溶液的吸光度。
结果如图2所示,经冠突散囊菌发酵获取的枸杞发酵液中的DPPH自由基清除率均明显高于未发酵的枸杞液,其中,发酵后第9天枸杞发酵液的DPPH自由基清除率高达75%,由此可见,经冠突散囊菌发酵获取的枸杞提取物其抗氧化能力显著提高,优于未发酵的枸杞液。
(2)ABTS清除率测定:取ABTS溶液2 mL加入到小试管中,加95%乙醇(或无水乙醇)1 mL,充分混合,在734 nm波长处测定其吸光度AABTS。依次分别加入100 μL不同发酵天数的枸杞发酵液,充分混合,静置反应30分钟后在734 nm波长处测定其吸光度A,以Vc为阳性对照。按照公式(2)计算不同天数发酵物ABTS自由基清除率:
式中:AABTS为对照组ABTS溶液的吸光度;A样品为加入样品溶液或Vc后溶液的吸光度。
结果如图3所示,经冠突散囊菌发酵获取的枸杞发酵液中的ABTS自由基清除率均明显高于未发酵的枸杞液,其中,发酵后第11天枸杞发酵液的ABTS自由基清除率高达76%。由此可见,经冠突散囊菌发酵获取的枸杞提取物其抗氧化能力显著提高,优于未发酵的枸杞液。
综上所述,本发明采用冠突散囊菌发酵枸杞制得枸杞发酵物,发酵工艺简单、周期短、操作温度低,枸杞发酵物清除DPPH、ABTS自由基的活性均优于未发酵的枸杞液,制备的枸杞发酵物具有显著的抗氧化活性,该发明为进一步合理利用枸杞资源奠定了基础,提高了枸杞的农业附加值,适用于食品、保健品、化妆品或药品等领域。
以上所述仅为本发明的优选实施方式,并非因此限制本发明的专利范围,凡是利用本发明说明书及附图内容所作的等效结构或等效流程变换,或直接或间接运用在其它相关的技术领域,均同理包括在本发明的专利保护范围内。
Claims (4)
1.一种具有抗氧化活性的枸杞发酵物,其特征在于,该枸杞发酵物通过冠突散囊菌发酵枸杞制备而成,其具有显著的抗氧化活性。
2.一种如权利要求1所述的枸杞发酵物的制备方法,其特征在于,该制备方法包括如下步骤:
(1)原料准备:取适量湿枸杞清洗备用;
(2)打浆:按质量比为1:20加入蒸馏水,用打浆器将枸杞的浆水打出,并搅拌均匀;
(3)高压灭菌:将步骤2中的均浆液体分装至三角瓶,每瓶200 ml,密封后进行高压灭菌,冷却备用;
(4)接种冠突散囊菌:将步骤3中的匀浆液体在无菌条件下按体积比1:400接种冠突散囊菌液;
(5)发酵:将步骤4中的液体密封、摇匀,置于震荡培养箱中进行发酵,发酵温度为28 °C,震荡培养箱的转速为200 r/min,发酵时间为1-15天;
(6)过滤:将步骤5中发酵后的发酵混合物过滤,得到液体枸杞发酵物。
3.根据权利要求2所述的枸杞发酵物的制备方法,其特征在于:所述步骤4中冠突散囊菌液的菌种密度为1×10 6/ml。
4.一种如权利要求1所述的枸杞发酵物在制备食品、保健品、化妆品或药品中的应用。
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