CN107279972A - 基于马鲛鱼活性多肽的复配物 - Google Patents
基于马鲛鱼活性多肽的复配物 Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Wood Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明公开了基于马鲛鱼活性多肽的复配物,成分及其重量份为:马鲛鱼活性多肽2~3份、1mol/L的氯化亚铁溶液0.5~1份、抗坏血酸0.1~0.3份、水90~100份。复配物的制备步骤包括:原料预处理、脱脂、一步酶解、二步酶解、脱色、脱腥、分离纯化、复配物配制。有益效果为:本发明复配物可通过氨基酸或肽的吸收通道被吸收,提高吸收效率,能高效快速地合成蛋白质,加快机体吸收金属离子的进程;利用率高,无金属异味,性质稳定,无毒副作用,具有改善缺铁性贫血的功能;采用两步酶解法制备活性肽,使得到的多肽分子量低,含量较高,无腥苦味,颜色较淡。
Description
技术领域
本发明涉及鱼类产品深加工技术领域,尤其是涉及基于马鲛鱼活性多肽的复配物。
背景技术
马鲛鱼(Scomberomorus niphonius)体形狭长,头及体背部蓝黑色,一般体长为25~50厘米、体重300~1000克。分布于北太平洋西部,我国产于黄海和渤海。马鲛鱼肉质细嫩洁白,糯软鲜爽,营养丰富,尾巴的味道特别好,物美价廉,素有"鲳鱼嘴,马鲛尾"之说。马鲛鱼生得浑身肥满,是一种经济价值较高的海产优质鱼类,每百克鱼肉含蛋白质19.1克,脂肪2.5克,并含多种维生素等。马鲛鱼胆固醇含量低,富含提高人脑智力的DHA元素和大量蛋白质、氨基酸以及钙、铁、钠等微量元素,还具有提神和防衰老等食疗功效,常食对治疗贫血、早衰、营养不良、产后虚弱和神经衰弱等症有一定辅助疗效。马鲛鱼肉多刺少,肉嫩味美,民间有"山上鹧鸪獐,海里马鲛鱼"的赞誉。
生物活性肽(Bioactive peptides)是指一类有特殊生理活性的肽类,一般由3到20个氨基酸组成,具有各种代谢和生理调节功能。肽的吸收机制要比氨基酸好,蛋白质经过蛋白酶酶解后,大部分以多肽形式被吸收,所以其营养价值要比游离态的氨基酸高。生物活性肽按来源可以分为陆地和海洋生物活性肽;按功能可以分为感官肽和生理功能肽,其中生理功能肽包括抑菌肽、抗氧化肽、降血压肽、降血脂肽等;按获取方式则可以分为天然存在的生物活性肽、人工合成的和酶解制备的生物活性肽。目前生物活性肽的高效制备主要采用酶解的方式,这种方法有许多优点,如条件温和、安全性高、成本低等,因而备受关注。利用海洋生物制备生产海洋生物活性肽已成为现在研究的热点,海洋中存在各种天然生物活性物质,作为海洋活性物质的重要组成部分天然海洋活性多肽依然成为未来发展的焦点。
现有技术如授权公告号为CN 103937864 B的中国发明专利,公开了一种从鳀鱼中分离的活性多肽,得到的抗神经细胞损伤多肽分子量为2,181Da;由天门冬氨酸、苏氨酸、谷氨酸、甘氨酸、丙氨酸、异亮氨酸、酪氨酸、苯丙氨酸、组氨酸、赖氨酸和精氨酸等十一种氨基酸组成,是一种新型的具有活性的多肽。该多肽可以显著提高PC12神经细胞的存活率,在终浓度为50μg/mL时,细胞存活率与损伤组比较提高了26.4%,有效地减轻了谷氨酸盐诱导的神经细胞损伤,但该制备方法比较复杂,不合适工业化生产。
发明内容
本发明的目的在于提供一种吸收效率高,无金属异味,性质稳定,无毒副作用,具有改善缺铁性贫血作用的基于马鲛鱼活性多肽的复配物。
本发明针对上述技术中提到的问题,采取的技术方案为:基于马鲛鱼活性多肽的复配物,成分及其重量份为:马鲛鱼活性多肽2~3份、1mol/L的氯化亚铁溶液0.5~1份、抗坏血酸0.1~0.3份、水90~100份。上述复配物是一种多肽-亚铁螯合物,可制成医用食品,能容易的透过肠粘膜而被肠道有效吸收,促进和提高了人体对铁元素的吸收利用,利用率高,对人体无毒副作用,无金属异味,性质稳定,具有改善缺铁性贫血,促进生长、延缓衰老及增强机体免疫力等生物及营养功能活性。
基于马鲛鱼活性多肽的复配物的制备方法,具体操作步骤为:
原料预处理:将马鲛鱼副产物切块,烘干,粉碎后过100~120目筛,2~6℃下密封保存,备用;
脱脂:将鱼粉以料液比为1:2~4加入石油醚,20~30℃下抽提5~7h脱脂,过滤,清水洗净,待石油醚挥发完全后烘干;
一步酶解:将脱脂后鱼粉按料液比为1:2~4加水,混合均匀,调节pH值至6~7,按0.5~1.5%加酶量添加木瓜蛋白酶,在52~75℃下酶解2~3h,灭酶,备用;
二步酶解:将脱脂一步酶解后鱼粉混合液,调节pH值至8~9,按1~2%加酶量添加碱性蛋白酶,在50~60℃下酶解2~3h,灭酶,离心,酶解液备用,因为碱性蛋白酶水解能力强,但水解位点的活性氨基酸较少,得到的大分子肽段多,经离心得到的多肽得率少,不宜作为第一步酶解,而木瓜蛋白酶的水解位点的活性氨基酸较多,故应先用木瓜蛋白酶水解,再用碱性蛋白酶作为第二步深度水解,既保证得到的水解液多肽分子量低,同时含量较高;
脱色、脱腥:将酶解液pH调节至3.5~4.5,加入0.3~0.8wt%的活性炭,在40~50℃下搅拌40~50min,经2~3µm微滤膜微滤,微滤液备用,活性炭具有发达的空隙结构,比表面积大,当被脱除的有色物质分子直径小于或等于活性炭孔的入口直径时,可被活性炭吸附,起到脱色、脱腥作用;
分离纯化:将微滤液pH调节至5.5~6.5,浓度为2~3%,在压力为0.3~0.8MPa下纳滤90~110min,超滤收集分子量为1~3kDa的肽段液,冷冻干燥,即得马鲛鱼活性肽,该活性多肽中含有一种氨基酸序列为HSHACASYSRCYCRCLRCRVLHPGCVNCSR的活性肽,该活性肽带有高浓度的负电荷,可通过多肽的氨基、亚氨基、羰基或羧基以共价键的形式与金属离子发生配位形成螯合物,金属离子借助肽形成有机态,避免了肠道吸收中某些物质对金属离子的沉淀或吸附,可通过氨基酸或肽的吸收通道被吸收,提高吸收效率,能高效快速地合成蛋白质,加快机体吸收金属离子的进程;
复配物配制:按配方称取马鲛鱼活性多肽、1mol/L的氯化亚铁溶液、抗坏血酸、水,在pH为6~7,温度为25~35℃下超声螯合25~35min,即得基于马鲛鱼活性多肽的复配物,该复配物是一种多肽-亚铁螯合物,可制成医用食品,能容易的透过肠粘膜而被肠道有效吸收,促进和提高了人体对铁元素的吸收利用,利用率高,对人体无毒副作用,无金属异味,性质稳定,具有改善缺铁性贫血,促进生长、延缓衰老及增强机体免疫力等生物及营养功能活性。
与现有技术相比,本发明的优点在于:1)本发明复配物是一种多肽-亚铁螯合物,可通过氨基酸或肽的吸收通道被吸收,提高吸收效率,能高效快速地合成蛋白质,加快机体吸收金属离子的进程;2)利用率高,无金属异味,性质稳定,无毒副作用,具有改善缺铁性贫血,促进生长、延缓衰老及增强机体免疫力等生物及营养功能活性;3)该制备方法采用两步酶解法制备活性肽,使得到的多肽分子量低,含量较高,无腥苦味,颜色较淡。
具体实施方式
下面通过实施例对本发明方案作进一步说明:
实施例1:
基于马鲛鱼活性多肽的复配物,成分及其重量份为:马鲛鱼活性多肽2.3份、1mol/L的氯化亚铁溶液0.8份、抗坏血酸0.2份、水95份。
基于马鲛鱼活性多肽的复配物的制备方法,具体操作步骤为:
1)原料预处理:将马鲛鱼副产物切块,在90℃烘箱中烘干,粉碎后过120目筛,5℃下密封保存,备用;
2)脱脂:将鱼粉以料液比为1:3加入石油醚,23℃下抽提6h脱脂,过滤,清水洗净,待石油醚挥发完全后烘干;
3)一步酶解:将脱脂后鱼粉按料液比为1:3加水,混合均匀,调节pH值至6.5,按0.8%加酶量添加木瓜蛋白酶,在55℃下酶解3h,110℃下灭酶17min,备用;
4)二步酶解:将脱脂一步酶解后鱼粉混合液,调节pH值至8,按1.6%加酶量添加碱性蛋白酶,在55℃下酶解2h,110℃下灭酶17min,4000r/min下离心15min,去除沉淀物质,留酶解液备用,因为碱性蛋白酶水解能力强,但水解位点的活性氨基酸较少,得到的大分子肽段多,经离心得到的多肽得率少,不宜作为第一步酶解,而木瓜蛋白酶的水解位点的活性氨基酸较多,故应先用木瓜蛋白酶水解,再用碱性蛋白酶作为第二步深度水解,既保证得到的水解液多肽分子量低,同时含量较高;
5)脱色、脱腥:将酶解液pH调节至4,加入0.6wt%的活性炭,在48℃下搅拌50min,经2µm微滤膜微滤掉活性炭,微滤液备用,活性炭具有发达的空隙结构,比表面积大,当被脱除的有色物质分子直径小于或等于活性炭孔的入口直径时,可被活性炭吸附,起到脱色、脱腥作用;
6)分离纯化:将微滤液pH调节至6,浓度为2%,在压力为0.8MPa下纳滤100min,超滤收集分子量为1~3kDa的肽段液,冷冻干燥,即得马鲛鱼活性肽,该活性多肽中含有一种氨基酸序列为HSHACASYSRCYCRCLRCRVLHPGCVNCSR的活性肽,该活性肽带有高浓度的负电荷,可通过多肽的氨基、亚氨基、羰基或羧基以共价键的形式与金属离子发生配位形成螯合物,金属离子借助肽形成有机态,避免了肠道吸收中某些物质对金属离子的沉淀或吸附,可通过氨基酸或肽的吸收通道被吸收,提高吸收效率,能高效快速地合成蛋白质,加快机体吸收金属离子的进程;
7)复配物配制:按配方称取马鲛鱼活性多肽、1mol/L的氯化亚铁溶液、抗坏血酸、水,在pH为6.8,温度为30℃下超声螯合28min,即得基于马鲛鱼活性多肽的复配物,该复配物是一种多肽-亚铁螯合物,可制成医用食品,能容易的透过肠粘膜而被肠道有效吸收,促进和提高了人体对铁元素的吸收利用,利用率高,对人体无毒副作用,无金属异味,性质稳定,具有改善缺铁性贫血,促进生长、延缓衰老及增强机体免疫力等生物及营养功能活性。
实施例2:
基于马鲛鱼活性多肽的复配物,成分及其重量份为:马鲛鱼活性多肽3份、1mol/L的氯化亚铁溶液0.7份、抗坏血酸0.1份、水95份。
基于马鲛鱼活性多肽的复配物的制备方法,具体操作步骤为:
1)原料预处理:将马鲛鱼副产物切块,烘干,粉碎后过100目筛,3℃下密封保存,备用;
2)脱脂:将鱼粉以料液比为1:3加入石油醚,25℃下抽提5h脱脂,过滤,清水洗净,待石油醚挥发完全后烘干,
3)一步酶解:将脱脂后鱼粉按料液比为1:3加水,混合均匀,调节pH值至7,按1.1%加酶量添加木瓜蛋白酶,在55℃下酶解2.5h,灭酶,备用;
4)二步酶解:将脱脂一步酶解后鱼粉混合液,调节pH值至8.5,按1.3%加酶量添加碱性蛋白酶,在57℃下酶解2h,灭酶,离心,酶解液备用,因为碱性蛋白酶水解能力强,但水解位点的活性氨基酸较少,得到的大分子肽段多,经离心得到的多肽得率少,不宜作为第一步酶解,而木瓜蛋白酶的水解位点的活性氨基酸较多,故应先用木瓜蛋白酶水解,再用碱性蛋白酶作为第二步深度水解,既保证得到的水解液多肽分子量低,同时含量较高;
5)脱色、脱腥:将酶解液pH调节至4,加入0.5wt%的活性炭,在45℃下搅拌45min,经3µm微滤膜微滤,微滤液备用,活性炭具有发达的空隙结构,比表面积大,当被脱除的有色物质分子直径小于或等于活性炭孔的入口直径时,可被活性炭吸附,起到脱色、脱腥作用;
6)分离纯化:将微滤液pH调节至5.5,浓度为3%,在压力为0.5MPa下纳滤90min,超滤收集分子量为1~3kDa的肽段液,冷冻干燥,即得马鲛鱼活性肽,该活性多肽中含有一种氨基酸序列为HSHACASYSRCYCRCLRCRVLHPGCVNCSR的活性肽,该活性肽带有高浓度的负电荷,可通过多肽的氨基、亚氨基、羰基或羧基以共价键的形式与金属离子发生配位形成螯合物,金属离子借助肽形成有机态,避免了肠道吸收中某些物质对金属离子的沉淀或吸附,可通过氨基酸或肽的吸收通道被吸收,提高吸收效率,能高效快速地合成蛋白质,加快机体吸收金属离子的进程;
7)复配物配制:按配方称取马鲛鱼活性多肽、1mol/L的氯化亚铁溶液、抗坏血酸、水,在pH为6.5,温度为33℃下超声螯合30min,即得基于马鲛鱼活性多肽的复配物,该复配物是一种多肽-亚铁螯合物,可制成医用食品,能容易的透过肠粘膜而被肠道有效吸收,促进和提高了人体对铁元素的吸收利用,利用率高,对人体无毒副作用,无金属异味,性质稳定,具有改善缺铁性贫血,促进生长、延缓衰老及增强机体免疫力等生物及营养功能活性。
本发明的操作步骤中的常规操作为本领域技术人员所熟知,在此不进行赘述。
以上所述的实施例对本发明的技术方案进行了详细说明,应理解的是以上所述仅为本发明的具体实施例,并不用于限制本发明,凡在本发明的原则范围内所做的任何修改、补充或类似方式替代等,均应包含在本发明的保护范围之内。
SEQUENCE LISTING
<110> 乐清益昌食品技术有限公司
<120> 基于马鲛鱼活性多肽的复配物
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<170> PatentIn version 3.5
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<213> 人工合成
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His Ser His Ala Cys Ala Ser Tyr Ser Arg Cys Tyr Cys Arg Cys Leu
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Claims (10)
1.基于马鲛鱼活性多肽的复配物,其特征在于:所述的复配物的成分及其重量份为:马鲛鱼活性多肽2~3份、1mol/L的氯化亚铁溶液0.5~1份、抗坏血酸0.1~0.3份、水90~100份。
2.根据权利要求1所述的基于马鲛鱼活性多肽的复配物,其特征在于:所述的活性多肽中含有一种氨基酸序列为HSHACASYSRCYCRCLRCRVLHPGCVNCSR的活性肽。
3.根据权利要求1所述的基于马鲛鱼活性多肽的复配物,其特征在于:所述的复配物的制备方法的步骤为:
1)原料预处理:将马鲛鱼副产物切块,烘干,粉碎后筛,备用;
2)脱脂:将鱼粉中加入石油醚,脱脂,过滤,清水洗净,烘干;
3)一步酶解:将脱脂后鱼粉加水,混合均匀,加入木瓜蛋白酶酶解,灭酶,备用;
4)二步酶解:向一步酶解中加入碱性蛋白酶酶解,灭酶,离心,备用;
5)脱色、脱腥:将酶解液加入活性炭,搅拌,微滤,备用;
6)分离纯化:将微滤液纳滤,超滤,冷冻干燥,即得马鲛鱼活性肽;
7)复配物配制:按配方称取配料,混合均匀,螯合。
4.根据权利要求3所述的基于马鲛鱼活性多肽的复配物的制备方法,其特征在于:所述的脱脂步骤中鱼粉与石油醚的料液比为1:2~4,脱脂温度为20~30℃,时间为5~7h。
5.根据权利要求3所述的基于马鲛鱼活性多肽的复配物的制备方法,其特征在于:所述的一步酶解步骤中鱼粉与水的料液比为1:2~4,加酶量为0.5~1.5%,pH为6~7,温度为52~75℃,时间为2~3h。
6.根据权利要求3所述的基于马鲛鱼活性多肽的复配物的制备方法,其特征在于:所述的二步酶解步骤中加酶量为1~2%,酶解pH为8~9,温度为50~60℃,时间为2~3h。
7.根据权利要求3所述的基于马鲛鱼活性多肽的复配物的制备方法,其特征在于:所述的脱色、脱腥步骤中:活性炭添加量为0.3~0.8wt%,pH为3.5~4.5,温度为40~50℃,时间为40~50min。
8.根据权利要求3所述的基于马鲛鱼活性多肽的复配物的制备方法,其特征在于:所述的脱色、脱腥步骤中微滤膜为2~3µm。
9.根据权利要求3所述的基于马鲛鱼活性多肽的复配物的制备方法,其特征在于:所述的分离纯化步骤中纳滤的pH为5.5~6.5,浓度为2~3%,压力为0.3~0.8MPa,时间为90~110min。
10.根据权利要求3所述的基于马鲛鱼活性多肽的复配物的制备方法,其特征在于:所述的复配物配制步骤中螯合pH为6~7,温度为25~35℃,时间为25~35min。
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