CN110407917A - 一种促进乳腺酪蛋白合成的章鱼寡肽及其制备方法与应用 - Google Patents
一种促进乳腺酪蛋白合成的章鱼寡肽及其制备方法与应用 Download PDFInfo
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Abstract
本发明公开了一种促进乳腺酪蛋白合成的章鱼寡肽及其制备方法与应用。所述的章鱼寡肽,包含以下氨基酸序列MGDVLNF,MGLAGPR,EAPLMHV和TEAPLMHV中的一种或二种以上。该章鱼寡肽能显著促进乳腺上皮细胞HC11生长,提高乳腺上皮细胞HC11的β‑酪蛋白的表达水平,比空白对照组提高了150%以上。结果表明该章鱼寡肽在产后妇女促进泌乳,提高乳汁质量的功能性营养食品中具有很好的应用前景。
Description
技术领域
本发明属于功能营养食品领域,具体涉及一种促进乳腺酪蛋白合成的章鱼寡肽及其制备方法与应用。
背景技术
母乳是婴儿出生后最理想的天然食品,无论在营养学和免疫学等方面母乳喂养都具有配方代乳品无法比拟的优点。世界卫生组织和联合国儿童基金会联合制定的《婴幼儿喂养全球战略》明确指出在最初6个月应对婴儿进行纯母乳喂养。但联合儿童基金会调查显示2012-2014年我国6月龄内婴儿纯母乳喂养率仅为28%,且3年内没有增加,与《中国儿童发展纲要(2011—2020)》提到的50%的目标还有较大差距。由于多种因素导致的产后妇女泌乳延迟、泌乳量不足等问题,是限制母乳喂养广泛实施的最主要因素。利用传统中药进行催乳,虽可见成效,但中药成分可能通过乳汁传递于新生儿,存在一定风险。
许多研究表明:血液中的必需氨基酸不能满足乳蛋白合成需要,血液中的寡肽可以弥补乳腺中氨基酸不足导致的乳蛋白合成下降,且寡肽促进乳蛋白合成的效率比游离氨基酸更高。乳腺对各种氨基酸的摄取量存在着很大差异,组氨酸(His),赖氨酸(Lys),蛋氨酸(Met),苯丙氨酸(Phe),酪氨酸(Tyr)和苏氨酸(Thr)为主要限制性氨基酸,乳腺对这些氨基酸的吸收效率高。另外Leu和精氨酸(Arg)还可作为功能性氨基酸促进乳腺上皮细胞酪蛋白的表达,调控乳蛋白合成。因此,含有这些氨基酸的肽段对酪蛋白合成可能具有更高的效率。目前没有关于从章鱼中提取分离寡肽并应用到产后功能食品的开发上的相关报道。
发明内容
本发明的第一个目的是提供一种促进乳腺酪蛋白合成的章鱼寡肽,所述的章鱼寡肽可以作为功能因子应用于产后功能食品的开发。
所述的促进乳腺酪蛋白合成的章鱼寡肽,包含以下氨基酸序列Met-Gly-Asp-Val-Leu-Asn-Phe(MGDVLNF),Met-Gly-Leu-Ala-Gly-Pro-Arg(MGLAGPR),Glu-Ala-Pro-Leu-Met-His-Val(EAPLMHV)和Thr-Glu-Ala-Pro-Leu-Met-His-Val(TEAPLMHV)中的一种或二种以上。
所述的促进乳腺酪蛋白合成的章鱼寡肽的分子量为500~2500kDa,含7~26个氨基酸,其中含His,Lys,Met,Phe,Tyr,Thr,Leu,Arg的10个氨基酸以下的肽段占75%以上。
本发明的第二个目的是提供一种上述的促进乳腺酪蛋白合成的章鱼寡肽的制备方法,包括以下步骤:将鲜活章鱼洗净,匀浆绞碎,加入去离子水和蛋白酶,恒温水浴震荡进行酶解,灭酶后离心取上清液,将上清液过滤,取滤液浓缩,用大孔吸附树脂DA201-C脱盐,浓缩,再用葡聚糖凝胶G-25分离,富集第一组峰样品,浓缩,冷冻干燥,得到章鱼寡肽。
所述的蛋白酶为碱性蛋白酶或中性蛋白酶,酶添加量为2000~4000 U/g。
所述的去离子水的加入量为1g/2~4mL。
所述的酶解温度为50~60℃,酶解pH为6.5~8.5,酶解时间为4~6h。
所述的灭酶是沸水浴灭酶10min。
所述的离心是在10000r/min,4℃低温离心30min。
所述的将上清液过滤是将上清液用5kDa超滤膜超滤。
本发明的第三个目的是提供上述的促进乳腺酪蛋白合成的章鱼寡肽在制备提高乳腺上皮细胞β-酪蛋白表达的制剂,或产后妇女促进泌乳,提高乳汁质量的制剂中的应用。
优选,所述的产后妇女促进泌乳,提高乳汁质量的制剂是产后妇女促进泌乳,提高乳汁质量的功能性营养食品。
本发明的第四个目的是提供一种提高乳腺上皮细胞β-酪蛋白表达的制剂或产后妇女促进泌乳,提高乳汁质量的制剂。所述的提高乳腺上皮细胞β-酪蛋白表达的制剂或产后妇女促进泌乳,提高乳汁质量的制剂均含有上述的促进乳腺酪蛋白合成的章鱼寡肽作为活性成分。优选,所述的产后妇女促进泌乳,提高乳汁质量的制剂是产后妇女促进泌乳,提高乳汁质量的功能性营养食品。
与现有技术相比,本发明具有如下优点和有益效果:
本发明首次从章鱼中分离得到具有促进泌乳,提高乳汁质量的寡肽组分,该寡肽组分能显著促进乳腺上皮细胞生长,提高酪蛋白的表达水平,与空白对照组比较,提高了150%以上,因此,所述的章鱼寡肽在产后妇女功能营养食品方面具有很好的开发价值和应用前景,可应用于制备产后妇女促进泌乳,提高乳汁质量的功能性营养食品。
附图说明
图1是章鱼肽OH的葡聚糖凝胶G-25的分离图谱。
图2是章鱼寡肽OH1的RP-HPLC图谱。
图3是章鱼寡肽OH1特征肽序列(MGLAGPR、MGDVLNF、EAPLMHV、TEAPLMHV)二级质谱图。
图4是不同浓度章鱼寡肽对乳腺上皮细胞增殖活力的影响。(单独浓度从左到右分别为章鱼肽OH、章鱼寡肽OH1、OH2、OH3、OH4)。
图5是章鱼寡肽对酪蛋白表达的影响。
具体实施方式
以下实施例是对本发明的进一步说明,而不是对本发明的限制。
实施例1
章鱼寡肽制备:将鲜活章鱼洗净,切小块用绞肉机匀浆绞碎,按料液比1g/2mL的比例加入去离子水,加入中性蛋白酶3000 U/g酶解,pH值为6.5,55℃恒温水浴震荡5h,沸水浴灭酶10min,10000r/min,4℃低温离心30min,取上清液,将上清液用5kDa超滤膜超滤,取滤液浓缩,随后用大孔吸附树脂DA201-C脱盐,浓缩,再用葡聚糖凝胶G-25分离,蒸馏水洗脱,分离谱图如图1所示,富集第一组峰样品(OH1),浓缩,冷冻干燥获得章鱼寡肽。
相应的,上述步骤中,用大孔吸附树脂DA201-C脱盐,浓缩得到的章鱼肽记为OH,再用葡聚糖凝胶G-25分离,蒸馏水洗脱,富集第一、第二、第三、第四组峰样品,浓缩,冷冻干燥获得的章鱼肽OH分离组分分别记为章鱼寡肽OH1、OH2、OH3、OH4。
实施例2
章鱼寡肽制备:将鲜活章鱼洗净,切小块用绞肉机匀浆绞碎,按料液比1g/3mL的比例加入去离子水,用NaOH调pH值为8.5,加入碱性蛋白酶2000 U/g酶解,60℃恒温水浴震荡6h,沸水浴灭酶10min,10000r/min,4℃低温离心30min,取上清液,将上清液用5kDa超滤膜超滤,取滤液浓缩,随后用大孔吸附树脂DA201-C脱盐,浓缩,再用葡聚糖凝胶G-25分离,蒸馏水洗脱,富集第一组峰样品(OH1),浓缩,冷冻干燥获得章鱼寡肽。
实施例3
章鱼寡肽制备:将鲜活章鱼洗净,切小块用绞肉机匀浆绞碎,按料液比1g/4mL的比例加入去离子水,用NaOH调pH值为8.0,加入碱性蛋白酶4000 U/g酶解,50℃恒温水浴震荡4h,沸水浴灭酶10min,10000r/min,4℃低温离心30min,取上清液,将上清液用5kDa超滤膜超滤,滤过液浓缩,随后用大孔吸附树脂DA201-C脱盐,浓缩,再用葡聚糖凝胶G-25分离,蒸馏水洗脱,富集第一组峰样品(OH1),浓缩,冷冻干燥获得章鱼寡肽。
实施例4
章鱼寡肽的氨基酸分析:将章鱼寡肽OH1(实施例1制得)分别经过酸水解和碱水解后,使用氨基酸自动分析仪分析游离氨基酸和水解后氨基酸组成。
结果显示:章鱼寡肽OH1总蛋白含量为79.03%,其中游离氨基酸仅含1.93%,而肽含量占到77.10%,含量较高的氨基酸有甘氨酸(Gly)、脯氨酸(Pro)、谷氨酸(Glu)、天冬氨酸(Asp)、精氨酸(Arg)、异亮氨酸(Ile)、亮氨酸(Leu)和赖氨酸(Lys)。
实施例5
章鱼寡肽的结构鉴定:
(1)将章鱼寡肽OH1(实施例1制得)进一步经安捷伦高效液相色谱分离制备,选用C18半制备色谱柱(250×4.6mm,0.5μm),流动相A:水(含0.1%v/v三氟乙酸);流动相B:乙腈(含0.1%v/v三氟乙酸),样品浓度为100mg/mL,上样量100μL,检测波长220nm,洗脱条件:0-7min,94%A;7-10min 85%-94%A;10-40min,45%-85%A。每次洗脱结束后平衡色谱柱20min,按图2收集各组分(F1-F6),多次上样,收集冻干。
(2)采用LC-ESI-MS/MS进行结构鉴定,各个样品组分(F1-F6)复溶于水,采用C18分析色谱柱(250×10mm,0.5μm),上样量为20μL,在高效液相色谱-高分辨质谱联用仪中对各组分进行多肽结构鉴定。柱子洗脱方式采用等度和梯度洗脱,其中流动相A为含有0.1%v/v乙酸的超纯水,流动相B为含有0.1%v/v乙酸的甲醇,各组分洗脱条件如下,F1和F2:0-5min,94%A;5-35min,94%-75%A,F3:0-5min,88%A;5-35min,88%-85%A,F4:0-5min,83%A;5-35min,83%-50%A,F5:0-5min,70%A;5-35min,70%-50%A,F6:0-5min,65%A,5-35min,65%-53%A;以220nm波长的吸光度进行检测控制。质谱端采用电喷雾离子源,正离子扫描模式,雾化气和干燥气为高纯氮气,扫描范围从200m/z到2000m/z,并根据一级质谱信号强度选择母离子进行碰撞诱导裂解获得二级质谱数据,通过Mascot数据库进行搜索分析,得到章鱼寡肽OH1的氨基酸序列。
结果共鉴定112条肽段,分子量主要分布在500-2500kDa之间,含7到26个氨基酸。筛选出含有His,Lys,Met,Phe,Tyr,Thr,Leu,Arg的10个氨基酸以下的肽段共88条,根据乳腺对氨基酸利用,His,Lys,Met,Phe,Tyr和Thr为主要限制性氨基酸,Leu,Arg为功能氨基酸,且疏水性较大的肽段更有利于寡肽的转运等报道,其中肽段MGDVLNF(Met-Gly-Asp-Val-Leu-Asn-Phe),MGLAGPR(Met-Gly-Leu-Ala-Gly-Pro-Arg),Glu-Ala-Pro-Leu-Met-His-Val(EAPLMHV)和Thr-Glu-Ala-Pro-Leu-Met-His-Val(TEAPLMHV)(二级质谱图如图3所示)可能对酪蛋白表达具有较大贡献。
实施例6
章鱼寡肽对乳腺上皮细胞生长及酪蛋白表达的影响
(1)细胞培养:选用小鼠乳腺上皮细胞系HC11细胞。细胞株于37℃恒温水箱中快速复苏,洗涤离心数次。于含2mmol/LL-谷氨酰胺,10%的胎牛血清,10ng/mL表皮生长因子和5μg/mL胰岛素的RPMI1640培养基中扩大培养。为诱导β-酪蛋白的合成,随后在RPMI1640基础培养基中补充10%胎牛血清,5μg/mL胰岛素,10ng/mL表皮生长因子,1μg/mL氢化可的松和1μg/mL催乳素。
(2)MTT法测定乳腺上皮细胞HC11的增殖实验:分别为空白对照组(control)、章鱼肽组(OH,实施例1制得)、章鱼寡肽组(OH1-OH4,实施例1制得)。采用MTT法检测乳腺上皮细胞的增殖活性。用含10%胎牛血清,5μg/mL胰岛素,10ng/mL表皮生长因子,1μg/mL氢化可的松和1μg/mL催乳素的RPMI1640培养液将细胞制成单细胞悬液,调整活细胞浓度为1×105个/mL加于96孔培养板,每孔100μL,于CO2培养箱中培养48h,向96孔板内加入含有不同浓度的章鱼寡肽的培养液,设空白对照组,培养2天后吸出孔内培养液,加入MTT溶液,并将其放入细胞培养箱内孵育,弃上清,每孔加入150μL的DMSO,振荡15min后,酶标仪560nm测定OD。按下列公式计算存活率,结果如图4所示。
存活率%=加药孔平均OD值/对照孔平均OD值×100%。
采用MTT法检测乳腺上皮细胞的增殖活性,与空白对照比较,章鱼肽OH,章鱼肽OH的分离组分OH1、OH2对细胞增殖具有一定促进作用,在浓度为25μg/mL作用下,三者都具有较好的活性。
(3)WB法检测β-酪蛋白的表达量:用含10%胎牛血清,5μg/mL胰岛素,10ng/mL表皮生长因子,1μg/mL氢化可的松和1μg/mL催乳素的RPMI1640培养液将细胞制成单细胞悬液,调整活细胞浓度为1×105个/mL加于6cm培养皿中,每孔3mL,于CO2培养箱中培养48h,向6cm培养皿内加入含有不同浓度的章鱼寡肽的培养液,设细胞对照组,培养2天后吸出孔内培养液。
①提取总蛋白:将待处理细胞用PBS清洗2次;随后向培养皿内加入250μL蛋白提取匀浆液;在冰上将细胞用细胞铲全部刮下,然后用移液枪将细胞连同匀浆液吸入1.5mL灭菌EP管中,EP管于漩涡振荡仪上高速振荡30s,迅速插入冰中冷却,重复上述过程,EP管内无明显浑浊物说明细胞裂解完全,可用于后续实验。蛋白样品分装后于-80℃保存备用。
②BCA蛋白定量:按BCA蛋白定量试剂盒检测。
③10%SDS-PAGE电泳:配制分离胶、浓缩胶,灌胶凝固。在蛋白样品中加入适量SDS-PAGE蛋白上样缓冲液(3×)和灭菌ddH2O,将蛋白样品连同上样缓冲液在PCR仪中95℃或沸水浴中变性10min,待蛋白充分变性后,将其取出在室温冷却,将冷却后的20μg蛋白样品上样,先恒压70 V电泳30min,后转恒压120V继续电泳,直至溴酚蓝刚好跑出凝胶下端,观察彩虹预染蛋白分子量marker位置,预计目的蛋白已经被分离后可停止电泳。
④转膜:在4℃进行转膜,设定转膜电流为450mA,转膜时间根据分子量不同设定在45~90min。
⑤封闭:将PVDF膜有蛋白一侧朝下放入预先准备好的1×TBST中清洗1次,5min后,将PVDF膜放入含5%脱脂奶粉或5%BSA的封闭液中封闭1h。
⑥一抗孵育:封闭结束后,吸去封闭液,加1×TBST清洗3次,每次10min。清洗后立即加入按比例稀释好的对应一抗,4℃在摇床缓慢摇动孵育过夜。一抗孵育结束,回收一抗,用1×TBST清洗PVDF膜3次,每次10min。
⑦二抗孵育:按照二抗稀释比例准备所需二抗,待孵育前放4℃解冻。TBST清洗后立即加入二抗稀释液,室温摇床缓慢摇动孵育1h,回收二抗稀释液。用1×TBST清洗PVDF膜3次,每次10min。
⑧蛋白检测:PVDF膜用化学发光液进行显色,拍照。数据处理用Image Pro-plμs软件进行灰度值分析。选用β-actin作为内参。结果如图5所示。
图5中,章鱼肽OH和章鱼寡肽OH1对HC11细胞内β-酪蛋白表达具有较好的促进作用,与空白对照组相比,分别提高了137.38%和161.65%。
以上仅是本发明的优选实施方式,应当指出的是,上述优选实施方式不应视为对本发明的限制,本发明的保护范围应当以权利要求所限定的范围为准。对于本技术领域的普通技术人员来说,在不脱离本发明的精神和范围内,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (10)
1.一种促进乳腺酪蛋白合成的章鱼寡肽,其特征在于,包含以下氨基酸序列Met-Gly-Asp-Val-Leu-Asn-Phe,Met-Gly-Leu-Ala-Gly-Pro-Arg,Glu-Ala-Pro-Leu-Met-His-Val和Thr-Glu-Ala-Pro-Leu-Met-His-Val中的一种或二种以上。
2.根据权利要求1所述的促进乳腺酪蛋白合成的章鱼寡肽,其特征在于,所述的章鱼寡肽分子量为500~2500kDa,含7~26个氨基酸,其中含His,Lys,Met,Phe,Tyr,Thr,Leu,Arg的10个氨基酸以下的肽段占75%以上。
3.一种权利要求1所述的促进乳腺酪蛋白合成的章鱼寡肽的制备方法,其特征在于,包括以下步骤:将鲜活章鱼洗净,匀浆绞碎,加入去离子水和蛋白酶,恒温水浴震荡进行酶解,灭酶后离心取上清液,将上清液过滤,取滤液浓缩,用大孔吸附树脂DA201-C脱盐,浓缩,再用葡聚糖凝胶G-25分离,富集第一组峰样品,浓缩,冷冻干燥,得到章鱼寡肽。
4.根据权利要求3所述的促进乳腺酪蛋白合成的章鱼寡肽的制备方法,其特征在于,所述的蛋白酶为碱性蛋白酶或中性蛋白酶,酶添加量为2000~4000U/g。
5.根据权利要求3所述的促进乳腺酪蛋白合成的章鱼寡肽的制备方法,其特征在于,所述的去离子水的加入量为1g/2~4mL;所述的酶解温度为50~60℃,酶解pH为6.5~8.5,酶解时间为4~6h。
6.根据权利要求3所述的促进乳腺酪蛋白合成的章鱼寡肽的制备方法,其特征在于,所述的离心是在10000r/min,4℃低温离心30min;所述的将上清液过滤是将上清液用5kDa超滤膜超滤。
7.权利要求1所述的促进乳腺酪蛋白合成的章鱼寡肽在制备提高乳腺上皮细胞β-酪蛋白表达的制剂,或产后妇女促进泌乳,提高乳汁质量的制剂中的应用。
8.根据权利要求7所述的应用,其特征在,所述的产后妇女促进泌乳,提高乳汁质量的制剂是产后妇女促进泌乳,提高乳汁质量的功能性营养食品。
9.一种提高乳腺上皮细胞β-酪蛋白表达的制剂或产后妇女促进泌乳,提高乳汁质量的制剂,其特征在于,含有权利要求1所述的促进乳腺酪蛋白合成的章鱼寡肽作为活性成分。
10.根据权利要求9所述的制剂,其特征在于,所述的产后妇女促进泌乳,提高乳汁质量的制剂是产后妇女促进泌乳,提高乳汁质量的功能性营养食品。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113501861A (zh) * | 2021-08-01 | 2021-10-15 | 青海瑞肽生物科技有限公司 | 一种高分辨质谱筛选牦牛胶原蛋白特征片段的方法 |
CN114163494A (zh) * | 2021-11-10 | 2022-03-11 | 中国科学院南海海洋研究所 | 一种拮抗氧化应激损伤的章鱼抗氧化肽的制备方法和应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH1142054A (ja) * | 1997-07-29 | 1999-02-16 | Asahi Chem Ind Co Ltd | 小麦加工食品用改良剤 |
CN101791125A (zh) * | 2010-01-29 | 2010-08-04 | 中国科学院南海海洋研究所 | 一种促进产后妇女泌乳及恢复的功能营养品及其制备方法 |
CN106962941A (zh) * | 2017-03-22 | 2017-07-21 | 深圳太太药业有限公司 | 一种用于产后、哺乳调养的海洋膳食组合物及其制备方法 |
-
2019
- 2019-07-08 CN CN201910608720.9A patent/CN110407917B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH1142054A (ja) * | 1997-07-29 | 1999-02-16 | Asahi Chem Ind Co Ltd | 小麦加工食品用改良剤 |
CN101791125A (zh) * | 2010-01-29 | 2010-08-04 | 中国科学院南海海洋研究所 | 一种促进产后妇女泌乳及恢复的功能营养品及其制备方法 |
CN106962941A (zh) * | 2017-03-22 | 2017-07-21 | 深圳太太药业有限公司 | 一种用于产后、哺乳调养的海洋膳食组合物及其制备方法 |
Non-Patent Citations (4)
Title |
---|
BINGNA CAI等: "Identification of octopus peptide and its promotion of -casein synthesis in a mouse mammary epithelial cell line", 《J FOOD BIOCHEM.》 * |
PARK, YEUNG-BEOM等: "Characteristics of Angiotensin Converting Enzyme Inhibitory Peptides from Aroase AP10 Hydrolysate of Octopus", 《JOURNAL OF THE KOREAN SOCIETY OF FOOD SCIENCE AND NUTRITION》 * |
ZHENG FANG等: "The preservative potential of Octopus scraps peptides−Zinc chelate against Staphylococcus aureus: Its fabrication, antibacterial activity and action mode", 《FOOD CONTROL》 * |
贾晨等: "章鱼下脚料发酵液中多肽的分离及抗氧化活性的研究", 《青岛农业大学学报(自然科学版)》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113501861A (zh) * | 2021-08-01 | 2021-10-15 | 青海瑞肽生物科技有限公司 | 一种高分辨质谱筛选牦牛胶原蛋白特征片段的方法 |
CN114163494A (zh) * | 2021-11-10 | 2022-03-11 | 中国科学院南海海洋研究所 | 一种拮抗氧化应激损伤的章鱼抗氧化肽的制备方法和应用 |
CN114163494B (zh) * | 2021-11-10 | 2023-10-10 | 中国科学院南海海洋研究所 | 一种拮抗氧化应激损伤的章鱼抗氧化肽的制备方法和应用 |
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