CN110801413A - 一种小球藻发酵提取物及其制备方法 - Google Patents
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Abstract
本发明公开了一种小球藻发酵提取物及其制备方法。本发明利用植物乳杆菌等菌种的培养液对小球藻粉进行液体发酵培养,得发酵产物,发酵产物经过离心、过滤、除菌,得小球藻粉发酵提取物。所述方法反应条件温和,制备工艺简单,无添加任何化学成分。通过实验证明,上述方法制备的小球藻粉发酵提取物,富含蛋白质、粗多糖、总氨基酸和叶黄素等功效成分,具良好的抗自由基、改善皮肤刺激及抗衰老等功效,且生物安全性高,可广泛应用到的各类化妆品中。
Description
技术领域
本发明属于生物技术领域,具体涉及一种小球藻发酵提取物及其制备方法。
背景技术
小球藻(Chlorella vulgaris)绿藻纲,小球藻科。单细胞藻,常单生,也有多细胞聚集。地球上最早的生命之一,是一种高效的光合植物,以光合自养生长繁殖,分布极广。小球藻因具有很高的应用价值,与螺旋藻、栅藻、盐藻等微藻的开发利用一起构成了微藻生物技术,具有广阔的发展前景。
小球藻含有丰富的蛋白质,含量高达50~60%,可作为单细胞蛋白的一个重要来源,同时还含有人和动物所必需的多种氨基酸、膳食纤维、维生素等营养成分。小球藻还具有低脂肪、低糖、低热量等优点,是一种优质的绿色营养源食品,在食品和保健食品等行业有巨大的经济价值。其中,蛋白核小球藻是一种具有巨大经济价值的天然资源,被***粮农组织(FAO)确定为二十一世纪绿色营养源健康食品。小球藻中含有小球藻生长因子COF、多糖和糖蛋白等多种生物活性物质,在医药方面对抗肿瘤、增强免疫力、抗病毒感染等效果显著。小球藻含有叶绿素、叶黄素和虾青素等多种色素,而且小球藻被发现可以累积虾青素,有可能成为继雨生红球藻之后又一种优质的天然虾青素藻源。
目前关于小球藻应用的研究主要集中在食品、医药、能源及水产养殖等领域,而在化妆品领域应用的研究及报道尚少。
发明内容
针对上述问题,本发明提供一种小球藻发酵提取物及其制备方法。本发明利用植物乳杆菌等菌种的培养液对小球藻粉进行液体发酵培养,得发酵产物,发酵产物经过离心、过滤、除菌,得小球藻粉发酵提取物。上述方法制备的小球藻粉发酵提取物,富含蛋白质、粗多糖、总氨基酸和叶黄素等功效成分,具良好的抗自由基、改善皮肤刺激及抗衰老等功效,开拓了小球藻在护肤领域的新应用。
为实现上述目的,本发明采用如下技术方案:一种小球藻发酵提取物的制备方法,其特征是,具体包括如下步骤:
(1)备种:将保存好的菌种接种于种子培养基中,进行扩大培养得到菌悬液;所述菌种为植物乳杆菌、干酪乳酸杆菌、副干酪乳酸杆菌及乳酸乳球菌中的至少一种,优选植物乳杆菌;
(2)液体发酵培养:将步骤(1)培养好的菌悬液接种于含有小球藻粉和葡萄糖的发酵培养基中,28~32℃静置培养24~48h(至葡萄糖含量为0),得到发酵液;
(3)调节pH:将发酵液pH值调至5.5~7.5;
(4)离心、过滤、除菌:将步骤(3)中得到的发酵液离心除残渣,收集上清,然后经过滤、除菌即得小球藻发酵提取物。
进一步的,所述步骤(1)中种子培养基的成分及含量为:M17培养基、葡萄糖5~20g/l,115℃灭菌30min;28~32℃静置培养12~36h得菌悬液。
进一步的,所述步骤(2)中菌悬液接种量(体积比)为:10~20%。
进一步的,所述步骤(2)中发酵培养基的成分及含量为:小球藻粉5~15g/l、葡萄糖10~20g/l,其余为蒸馏水,115℃灭菌30min。
进一步的,所述步骤(3)中,用1mol/L NaOH溶液调节发酵液pH至5.5~7.5。
进一步的,所述步骤(4)中,离心功率为4000~8000r/min,离心10~30min。
进一步的,所述步骤(4)的过滤除菌为:经过1.2μm精细过滤纸板、0.22μm聚醚砜滤芯过滤除菌。
上述方法制备的小球藻发酵提取物,其主要功效成分及含量(小球藻粉的用量约为10g/1.1L发酵液)为:蛋白质≥500mg/100ml;粗多糖≥440mg/100ml;总氨基酸≥190mg/100ml;叶黄素≥240μg/100ml。
上述方法制备的小球藻发酵提取物具有良好的抗自由基、改善皮肤刺激及抗衰老等功效,可以以它为有效成分制备化妆品。
本发明的有益效果是:
1、本发明以小球藻粉为原料,利用微生物发酵技术将其活性成分充分释放了出来,制备的小球藻发酵提取物富含蛋白质、粗多糖、总氨基酸和叶黄素。这些活性成分共同作用使小球藻发酵提取物具有良好抗自由基、改善皮肤刺激及抗衰老等功效,开拓了小球藻在护肤领域的新应用。
2、本发明通过试验证明:
1)本发明的小球藻粉发酵提取物在4.5-40.5%(V/V)浓度下能够提高ABTS自由基的清除率,且呈剂量依赖性模型,当样品浓度大于等于4.5%(V/V)时,与阴性对照组相比具有统计学差异;
2)本发明的小球藻发酵产物在0.16-10.00%(V/V)浓度下作用48h,能够提高NHDFs细胞的存活率,且呈剂量依赖性模型,当样品浓度大于等于0.63%(V/V)时,与阴性对照组相比具有统计学差异;
3)本发明的小球藻发酵产物在10%(V/V)浓度下作用48h,可提升NHDFs细胞的I型胶原蛋白合成能力,高于阳性对照(50μg/ml抗坏血酸),且与阴性对照相比具有统计学差异。说明产品具有良好抗自由基、改善皮肤刺激及抗衰老等功效。
3、本发明反应条件温和,制备工艺简单高效,无任何化学成分添加,可实现工业化大规模生产。
附图说明
图1为小球藻发酵提取物对ABTS自由基的清除作用;其中**:P值<0.01;
图2为小球藻发酵提取物对NHDFs细胞相对活率的影响;其中*:P值<0.05,**:P值<0.01;
图3为小球藻发酵提取物对I型胶原蛋白相对含量的影响;其中**:P值<0.01。
具体实施方式
以下结合实施例对本发明作进一步的说明,应该说明的是,下述说明仅是为了解释本发明,并不对其内容进行限定。下述实施例中所使用的实验方法无特殊说明,均为常规方法。下述实施例中所使用的材料、试剂等,如无特殊说明,均可通过商业途径获得。下述实施例中所使用的菌种购于中国工业微生物菌种保藏管理中心(CICC)。
实施例1:小球藻发酵提取物的制备
采用如下方法制备得到小球藻发酵提取物:
(1)备种:将保存好的植物乳杆菌接种于种子培养基中,30℃静置培养24h,得菌悬液;
种子培养基成分及含量为:M17培养基、葡萄糖20g/l,115℃灭菌30min。
(2)液体发酵培养:将步骤(1)中培养好的菌悬液按体积比10%的接种量接种于发酵培养基中,30℃静置培养36h,葡萄糖含量为0,结束发酵,得发酵液;
发酵培养基成分及含量为:小球藻粉10g/l、葡萄糖15g/l,其余为蒸馏水,115℃灭菌30min。
(3)调节pH:用1mol/LNaOH溶液调节发酵液pH至6.0左右。
(4)离心、过滤、除菌:首先将步骤(3)中得到的发酵液离心,8000r/min,离心10min,除残渣收集上清;然后经过1.2μm精细过滤纸板、0.22μm聚醚砜滤芯过滤除菌即得小球藻发酵提取物。
对小球藻发酵提取物中的主要功效成分及含量进行检测。结果如下:
表1主要功效成分及含量
实施例2:小球藻发酵提取物的制备
采用如下方法制备得到小球藻发酵提取物:
(1)备种:将保存好的植物乳杆菌接种于种子培养基中,30℃静置培养28h,得菌悬液;
种子培养基成分及含量为:M17培养基、葡萄糖15g/l,115℃灭菌30min。
(2)液体发酵培养:将步骤(1)中培养好的菌悬液按体积比15%的接种量接种于发酵培养基中,30℃静置培养30h,葡萄糖含量为0,结束发酵,得发酵液;
发酵培养基成分及含量为:小球藻粉10g/l、葡萄糖12g/l,其余为蒸馏水,115℃灭菌30min。
(3)调节pH:用1mol/LNaOH溶液调节发酵液pH至6.0左右。
(4)离心、过滤、除菌:首先将步骤(3)中得到的发酵液离心,6000r/min,离心15min,除残渣收集上清;然后经过1.2μm精细过滤纸板、0.22μm聚醚砜滤芯过滤除菌即得小球藻发酵提取物。
实施例3:小球藻发酵提取物的制备
采用如下方法制备得到小球藻发酵提取物:
(1)备种:将保存好的植物乳杆菌接种于种子培养基中,30℃静置培养20h,得菌悬液;
种子培养基成分及含量为:M17培养基、葡萄糖10g/l,115℃灭菌30min。
(2)液体发酵培养:将步骤(1)中培养好的菌悬液按体积比20%的接种量接种于发酵培养基中,30℃静置培养38h,葡萄糖含量为0,结束发酵,得发酵液;
发酵培养基成分及含量为:小球藻粉10g/l、葡萄糖20g/l,其余为蒸馏水,115℃灭菌30min。
(3)调节pH:用1mol/LNaOH溶液调节发酵液pH至6.0左右。
(4)离心、过滤、除菌:首先将步骤(3)中得到的发酵液离心,5000r/min,离心20min,除残渣收集上清;然后经过1.2μm精细过滤纸板、0.22μm聚醚砜滤芯过滤除菌即得小球藻发酵提取物。
试验例1:小球藻发酵提取物ABTS自由基清除能力检测
本实施例旨在对实施例1中制备的小球藻发酵提取物进行抗自由基功效评价
将样品用蒸馏水配制成相应浓度(0.5、1.5、4.5、13.5、40.5%(v/v))的待测液,在96孔板中按照表2中各试剂添加量配制反应体系,混匀,每个浓度设置3个复孔,1个背景对照孔。
表2ABTS自由基清除试验反应体系
室温孵育2-6min后,于734nm下读取吸光度OD值,计算样品对ABTS自由基的清除率,根据标准差计算公式得出SD值。
清除率计算方法:
式中:C1——空白有ABTS体系吸光度值;
C2——空白无ABTS体系吸光度值;
T1——样品组有ABTS体系吸光度值;
T2——样品组无ABTS体系吸光度值。
检测结果如表3所示,样品(小球藻发酵提取物)在4.5-40.5%(V/V)浓度下能够提高ABTS自由基的清除率,且呈剂量依赖性模型,当样品浓度大于等于4.5%(V/V)时,与阴性对照组相比具有统计学差异(见附图1)。
表3ABTS自由基清除试验结果分析(Mean±SD)
试验例2:小球藻提取物改善皮肤刺激及抗衰老功效评价
本实施例旨在对实施例1中制备的小球藻提取物进行抗衰老功效评价。
1、体外人成纤维细胞增殖试验
用DMEM培养基将样品(小球藻发酵产物)配置成10%(V/V)溶液,经0.22μm滤膜过滤后,将样品溶液进行连续对倍稀释至0.08%(V/V),共8个梯度浓度。
表4各测试组浓度设计表
收集对数生长期细胞,按照细胞密度为8×103个/孔接种至96孔板,每孔含培养液100μl。在培养箱(37℃,5%CO2)中培养24h后,吸去培养基,用PBS缓冲液洗涤细胞两次,然后按照表4加入药物,每孔100μl,以未处理细胞作为阴性对照,每组设置3个重复孔。
加药后在培养箱(37℃,5%CO2)中继续培养48h,然后进行CCK-8细胞毒性试验,450nm下读取吸光度OD值。按照以下公式计算细胞存活率,根据标准差计算公式得出SD值,空白孔中仅添加细胞培养液。
检测结果如表5所示,样品(小球藻发酵产物)在0.16-10.00%(V/V)浓度下作用48h,能够提高NHDFs细胞的存活率,且呈剂量依赖性模型,当样品浓度大于等于0.63%(V/V)时,与阴性对照组相比具有统计学差异(见附图2)。
表5细胞活率检测结果分析(作用48h)
2、体外紧致抗皱功效评估(胶原蛋白I含量)
ELISA法检测Ⅰ型胶原蛋白含量。
收集对数生长期人皮肤成纤维细胞(NHDFs),按照细胞密度为6×104个/孔接种至12孔板。在培养箱(37℃,5%CO2)中培养24h后,根据细胞毒性结果,按照10%(v/v)浓度加入样品(小球藻发酵产物),以50μg/ml抗坏血酸(VC)作为阳性对照,以未处理细胞作为阴性对照,以反应体系试剂及溶剂作为溶剂对照,每组设置4个重复孔。
加药后在培养箱(37℃,5%CO2)中继续培养48h,收集培养液。采用Col I ELISA试剂盒进行培养液中Ⅰ型胶原蛋白含量的检测,计算Ⅰ型胶原蛋白相对溶剂对照的相对含量,根据标准差计算公式得出SD值。
检测结果如表6所示,样品(小球藻发酵产物)在10%(V/V)浓度下作用48h,可提升NHDFs细胞的I型胶原蛋白合成能力至阴性对照的4.43±0.34倍,高于阳性对照,且与阴性对照相比具有统计学差异。说明,样品在此浓度下具有一定的紧致抗皱功效(见附图3)。
表6 I型胶原蛋白相对含量结果分析
浓度(%,V/V) | I型胶原蛋白相对含量 | SD |
阴性对照 | 1.00 | 0.19 |
阳性对照 | 2.90 | 0.05 |
10 | 4.43 | 0.34 |
Claims (10)
1.一种小球藻发酵提取物的制备方法,其特征是,具体包括如下步骤:
(1)备种:将保存好的菌种接种于种子培养基中,进行扩大培养得到菌悬液;所述菌种为植物乳杆菌、干酪乳酸杆菌、副干酪乳酸杆菌及乳酸乳球菌中的至少一种;
(2)液体发酵培养:将步骤(1)培养好的菌悬液接种于含有小球藻粉和葡萄糖的发酵培养基中,28~32℃静置培养24~48h,得到发酵液;
(3)调节pH:将发酵液pH值调至5.5~7.5;
(4)离心、过滤、除菌:将步骤(3)中得到的发酵液离心除残渣,收集上清,然后经过滤、除菌即得小球藻发酵提取物。
2.如权利要求1所述的一种小球藻发酵提取物的制备方法,其特征是,所述步骤(2)中发酵培养基的成分及含量为:小球藻粉5~15g/l、葡萄糖10~20g/l,其余为蒸馏水。
3.如权利要求1所述的一种小球藻发酵提取物的制备方法,其特征是,所述步骤(1)中种子培养基的成分及含量为:M17培养基、葡萄糖5~20g/l。
4.如权利要求1所述的一种小球藻发酵提取物的制备方法,其特征是,所述步骤(1)28~32℃静置培养12~36h得菌悬液。
5.如权利要求1所述的一种小球藻发酵提取物的制备方法,其特征是,所述步骤(2)中菌悬液接种量,按体积比为:10~20%。
6.如权利要求1所述的一种小球藻发酵提取物的制备方法,其特征是,所述步骤(3)中,用1mol/L NaOH溶液调节发酵液pH至5.5~7.5。
7.如权利要求1所述的一种小球藻发酵提取物的制备方法,其特征是,所述步骤(4)中,离心功率为4000~8000r/min,离心10~30min。
8.如权利要求1所述的一种小球藻发酵提取物的制备方法,其特征是,所述步骤(4)的过滤、除菌为:经过1.2μm精细过滤纸板、0.22μm聚醚砜滤芯过滤除菌。
9.权利要求1-8中任意一项所述的方法制备的小球藻发酵提取物。
10.权利要求9所述的小球藻发酵提取物在制备抗自由基、改善皮肤刺激及抗衰老功效的化妆品方面的应用。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113907344A (zh) * | 2021-11-03 | 2022-01-11 | 新疆金正生物科技有限公司 | 一种荒漠小球藻液体发酵制备工艺 |
CN114317616A (zh) * | 2021-12-08 | 2022-04-12 | 广东塔尖生物科技有限公司 | 一种菌类发酵产物的制备工艺及化妆品 |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20060060601A (ko) * | 2004-11-30 | 2006-06-05 | 가부시끼가이샤 교또 에이요 가가꾸 겡뀨쇼 | 클로렐라 발효 식품의 제조 방법 |
US20110250178A1 (en) * | 2008-11-07 | 2011-10-13 | Solazyme ,Inc. | Cosmetic compositions comprising microalgal components |
KR101252132B1 (ko) * | 2012-06-08 | 2013-04-08 | 대상 주식회사 | 고-유단백 우유 생성을 위한 사료 첨가제 및 이를 이용한 비유 젖소 사료의 제조방법 |
CN104351901A (zh) * | 2014-11-11 | 2015-02-18 | 甘肃德福生物科技有限公司 | 一种益生菌发酵小球藻饮料及其制作方法 |
CN106265360A (zh) * | 2016-09-29 | 2017-01-04 | 中国海洋大学 | 一种含海藻发酵产物的洗面奶及其制备方法 |
CN107334726A (zh) * | 2017-08-31 | 2017-11-10 | 广州市科能化妆品科研有限公司 | 一种祛痘组合物及其制备方法和应用 |
CN108203729A (zh) * | 2017-12-28 | 2018-06-26 | 广州市科能化妆品科研有限公司 | 一种巨藻抗氧化肽的制备方法 |
CN108245479A (zh) * | 2017-09-19 | 2018-07-06 | 王亚东 | 一种含有乳双歧杆菌发酵活性提取物的面膜 |
CN109260063A (zh) * | 2018-11-09 | 2019-01-25 | 吴静 | 一种基于藻类提取物的抗氧化乳液配方 |
-
2019
- 2019-12-12 CN CN201911272612.5A patent/CN110801413A/zh active Pending
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20060060601A (ko) * | 2004-11-30 | 2006-06-05 | 가부시끼가이샤 교또 에이요 가가꾸 겡뀨쇼 | 클로렐라 발효 식품의 제조 방법 |
US20110250178A1 (en) * | 2008-11-07 | 2011-10-13 | Solazyme ,Inc. | Cosmetic compositions comprising microalgal components |
KR101252132B1 (ko) * | 2012-06-08 | 2013-04-08 | 대상 주식회사 | 고-유단백 우유 생성을 위한 사료 첨가제 및 이를 이용한 비유 젖소 사료의 제조방법 |
CN104351901A (zh) * | 2014-11-11 | 2015-02-18 | 甘肃德福生物科技有限公司 | 一种益生菌发酵小球藻饮料及其制作方法 |
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