CN107853620A - 一种海参肠卵中生物活性成分集成提取的工艺 - Google Patents
一种海参肠卵中生物活性成分集成提取的工艺 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
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- A—HUMAN NECESSITIES
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- A23L17/65—Addition of, or treatment with, microorganisms or enzymes
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- A—HUMAN NECESSITIES
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
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- A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
本发明公开了一种海参肠卵中生物活性成分集成提取的工艺,涉及生物科技技术领域。取新鲜或解冻海参肠卵加纯水,将调配料通过胶体磨磨成浆液;将浆液通过均质机得均质液;将均质液打入酶解罐中,加复合蛋白酶,搅拌酶解,灭酶,料液经管式离心机分离得水相及油相;净化液经有机膜进行分离;截留液浓缩经乙醇处理,沉淀脱水后干燥即得海参肠卵粗多糖;精制液减压蒸发浓缩,得浓缩液;将浓缩液喷雾干燥得海参肠卵低聚肽;油相经石油醚萃取所得有机相经旋转蒸发除尽石油醚获取海参粗卵油。酶解采用单酶系统筛选,复合蛋白酶复筛后确定其配方,低聚肽含量的提高,直接增加了产品的消化、吸收和利用率。增加了产品收率,提高了产品质量。
Description
技术领域
本发明涉及一种海参肠卵中生物活性成分集成提取的工艺,具体为生物科技技术领域。
背景技术
海参肠卵,俗称“海参籽”,在日本称为“海参花”。海参肠卵不仅含有与海参体壁同样丰富的营养成分,而且许多珍贵的关键营养含量更多,决定了海参肠卵的至尊价值。其中多糖含量是刺参体壁的3-4倍,钒的含量是刺参体壁的3倍;海参肠卵内还含有特有的成分——性腺色素(β-胡萝卜素、海胆紫酮、虾青素(也称虾红素、玉米黄质、角黄素以及一些未知色素)。在滋补、防治和辅助治疗三方面的价值,可谓千金难寻的“无价之宝”。
王共明(2013)研究:(1)仿刺参营养较均衡,其中干品粗蛋白含量达到50.72g/100g,粗多糖含量高达27.25g/100g,粗脂肪中多以磷脂形式存在。高效液相色谱分析仿刺参卵氨基酸组成,总氨基酸和必需氨基酸的总量分别为34.71%和17.62%,必需氨基酸和总氨基酸比值高达50.77%(远高于WHO和FAO规定EA/TA≥40%为优质蛋白源);(2)仿刺参卵的最佳酶解工艺为:以混合酶1.5%,45℃酶解1h,水解度达到77.11%;(3)仿刺参卵多糖经纯化后发现在190nm到200nm处有强烈单一吸收峰,分子质量大于200kDa。
赵鸿霞等(2010)研究,以水解度和肽得率为评价指标,从4种酶解筛选出木瓜蛋白酶为水解最佳用酶,其条件为温度52℃,pH5.5,底物浓度为8-4%,加酶量为2500u/g,时间4h,在此条件下水解度为20.51%。
杨利伟(2012)研究了中类胡萝卜素的组成和含量并优化了提取工艺。分光光度法定量分析表明海参卵中总类胡萝卜素的含量为113μg/g。;高效液相色谱法测定虾青素的含量23μg/g。,占总类胡萝卜素的20.3%。有机溶剂法提取工艺条件为:以乙酸乙酯为提取剂,提取温度40℃,提取时间3h,料液比1:10,类胡萝卜素含量可达62.3μg/g;超声提取的工艺为:超声功率为70W,超声时间40min,类胡萝卜素含量为70.1μg/g;皂化处理法提取工艺为:皂化用碱0.2M NaoH,加碱量2-4μg/g,皂化时间50min,温度40℃,类胡萝卜素含量为72.3μg/g;发酵法提取工艺为:微生物黑曲霉84g,接种量8%,发酵时间3d,类胡萝卜素含量可达85.3μg/g;酶法提取工艺条件:脂肪酶加量8%,35℃反应35min,类胡萝卜素含量可达92.6μg/g。
杨利伟等(2013)研究,海参卵中类胡萝卜素提取条件的优化,以乙酸乙酯为提取溶剂,二次提取,温度40℃,提取时间3h,料液比为1:12,类胡萝卜素提取质量分数为63.20μg/g。
张婕(2014)研究海参肠卵提取物对雌性小鼠生殖系统的影响作用,结果发现(1)水溶液超声辅助提取,液料比1.5倍,.30'C条件下超声25min,提取2h,提率为8.1%;有机溶剂提取,液料比10倍,30℃条件下提取3h,提率为20.3%;(2)海参花提取物通过提高小鼠血清中激素含量,增加脏器指数和子宫内膜厚度,同时增加卵泡数量,改善卵巢形态,保护小鼠的生殖系统;(3)海参花提取物通过增加总蛋白含量,提高SOD、GSH-Px的活性,降低MDA的活性,抑制脂质过氧化反应,可能是它抗氧化能力的机制;(4)海参花提取物可以通过抑制Bax、Caspase-3的表达,增加Bcl-2的表达,抑制细胞凋亡。
刘昕(2016)以仿刺参卵为原料提取多糖,在单因素实验的基础上,通过响应面分析优化双酶提取仿刺参卵多糖工艺,并对其体外抗氧化活性进行初步研究。结果表明:仿刺参卵多糖的最佳提取条件为复合蛋白酶与木瓜蛋白酶配比1∶2、加酶量2.78%、酶解温度60℃、酶解时间6h,在此条件下卵多糖的预测得率为8.84‰,实际得率为8.76‰;仿刺参卵多糖的自由基清除活性呈明显的剂量依赖关系,对DPPH自由基的IC50为1.71mg/mL,是一种良好的天然抗氧化剂。
综叙上述对海参肠卵的研究,仍有如下不足:
(1)对海参肠卵的酶解用酶多采用单酶或是双酶酶解,导致酶解收率偏低;
(2)对酶解产物没有进行分子量测定,因此无法评价该产品的消化吸收率。
(3)酶解液未经精制、脱腥,可能影响口感。
(4)对海参肠卵的研究只是对单一物质的提取,而没有在一条工艺上最大限度的将其活性物质综合考虑提取,造成海参肠卵资源的浪费。
发明内容
本发明的目的在于提供一种海参肠卵中生物活性成分集成提取的工艺,以解决上述背景技术存在的问题。
为实现上述目的,本发明提供如下技术方案:一种海参肠卵中生物活性成分集成提取的工艺包括:1、取新鲜或解冻海参肠卵加纯水,其中料液比1:2-4,搅拌分散;
2、将调配料通过胶体磨磨成浆液;
3、将浆液通过均质机,40-60MPa压力下均质,得均质液;
4、将均质液打入酶解罐中,加热至45-55℃为料液;
5、料液用10%NaOH调pH7.5-8.5;
6、加占蛋白总量1.0-5.0%的复合蛋白酶,搅拌酶解3.0-6.0h;
7、料液pH调至5.0-7.0;
8、料液升温加热至70-90℃,灭酶10-30min;
9、料液经管式离心机分离得水相及油相;
10、水相过孔径10-100μm陶瓷膜进一步净化;
11、净化液经截留分子量为800-1200Da的有机膜进行分离;
12、截留液浓缩至固含量≥25%,经3-6倍体积95%乙醇处理,沉淀经无水乙醇脱水后干燥即得海参肠卵粗多糖;或加入1-10%糊精作为助凝剂,搅拌溶解后喷雾干燥得海参肠卵粗多糖;
13、透过液加1-5%的β-环糊精,在50℃-70℃搅拌处理15min-60min进行包埋脱腥,脱腥液过颗粒活性炭柱进一步脱腥脱色;
14、过柱液用孔径10-100μm陶瓷膜脱炭,得澄清精制液;
15、精制液减压蒸发浓缩至固形物30%-40%,得浓缩液;
16、将浓缩液喷雾干燥得海参肠卵低聚肽;
17、油相经2-5倍体积石油醚反复萃取3-5次,萃取所得有机相经旋转蒸发除尽石油醚获取海参粗卵油。
与现有技术相比,本发明的有益效果是:a、海参肠卵用纯水调配,经胶体磨磨浆后,再过40-60MPa高压均质机处理成浆料,使海参肠卵100%破壁利于酶解。
b、酶解采用单酶系统筛选,复合蛋白酶复筛后确定其配方,使酶解收率达80%以上,产品中分子量<1000Da部分占总蛋白比例超过90%。
c、低聚肽含量的提高,直接增加了产品的消化、吸收和利用率。
d、增加了产品收率,提高了产品质量,降低了产品成本,大幅度提高产
品在市场上的竞争力。
d、本工艺采用集成提取工艺,一次投料,获取多种活性产物,最大限度
提高海参肠卵的利用率。
f、工艺经脱腥处理,产品没有海参肠卵特有的腥味,口感好。
附图说明
图1是海参肠卵中集成提取生物活性成分的工艺流程图。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1
取新鲜或解冻海参肠卵100g,加纯水200g,放入破壁机中打成浆料;将浆液放入烧杯,并放入水浴加热至45℃±1℃;用10%NaOH调料液pH7.5±0.1;加1.0%复合蛋白酶(占蛋白总量),搅拌酶解3.0h;料液pH调至5.5;升温加热至70℃,30min灭酶;料液4000rpm离心10min,获取水相及油相;水相加1%β-环糊精,50℃搅拌处理60min进行包埋脱腥,再加1%颗粒活性炭,50℃搅拌处理60min进一步脱腥脱色,过0.2μm微孔滤膜抽滤得精制液;精制液减压蒸发浓缩至固形物30-40%,得浓缩液;浓缩液经实验型喷雾干燥塔喷干得海参肠卵低聚肽和粗多糖混合物;油相经石油醚反复萃取3-4次,萃取所得有机相经蒸馏获取海参粗卵油。
实施例2
取新鲜或解冻海参肠卵20kg,加纯水60kg,搅拌分散;将调配料通过胶体磨磨成浆液;将浆液通过均质机,50MPa压力下均质,得均质液;将均质液打入酶解罐中,加热至50℃±1℃;料液用10%NaOH调pH8.0±0.1;加3.0%复合蛋白酶(占蛋白总量),搅拌酶解4.5h;料液pH调至6.0;升温加热至75℃,35min灭酶;料液经管式离心机分离得水相及油相;水相过孔径10-100μm陶瓷膜进一步净化;净化液经截留分子量为800Da的有机膜进行分离;截留液浓缩至固含量≥25%,经3倍体积95%乙醇处理,沉淀经无水乙醇脱水后真空干燥即得海参肠卵粗多糖;透过液加3%β-环糊精,60℃搅拌处理40min进行包埋脱腥,再通过颗粒活性炭柱,60℃处理40min,进一步脱色脱味精制;过柱液用孔径10-100μm陶瓷膜脱炭,得澄清精制液;精制液减压蒸发浓缩至固形物30-40%,得浓缩液;将浓缩液喷雾干燥得海参肠卵低聚肽;油相经2倍体积石油醚反复萃取3-4次,萃取所得有机相经旋转蒸发除尽石油醚获取海参粗卵油。
实施例3
取新鲜或解冻海参肠卵300kg,加纯水1200L,搅拌分散;将调配料通过胶体磨磨成浆液;将浆液通过均质机,55MPa压力下均质,得均质液;将均质液打入酶解罐中,加热至52℃±1℃;料液用10%NaOH调pH8.5±0.1;加5.0%复合蛋白酶(占蛋白总量),搅拌酶解6h;料液pH调至6.5;料液升温加热至80℃,10min灭酶;料液经管式离心机分离得水相及油相;水相过孔径10-100μm陶瓷膜进一步净化;净化液经截留分子量为1000Da的有机膜进行分离;截留液浓缩至固含量≥25%,加入8%糊精作为助凝剂,搅拌溶解后喷雾干燥得海参肠卵粗多糖;透过液加5%β-环糊精,70℃搅拌处理15min脱腥脱色,再通过颗粒活性炭柱,70℃处理15min,进一步脱色脱味精制;过柱液用孔径10-100μm陶瓷膜脱炭,得澄清精制液;精制液减压蒸发浓缩至固形物30-40%,得浓缩液;将浓缩液喷雾干燥得海参肠卵低聚肽;油相经4倍体积石油醚反复萃取4-5次,萃取所得有机相经旋转蒸发除尽石油醚获取海参粗卵油。
尽管已经示出和描述了本发明的实施例,对于本领域的普通技术人员而言,可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由所附权利要求及其等同物限定。
Claims (1)
1.一种海参肠卵中生物活性成分集成提取的工艺,其特征在于:具体的提取工艺步骤为:(1)、取新鲜或解冻海参肠卵加纯水,其中料液比为1:2-4,搅拌分散;
(2)、将调配料通过胶体磨磨成浆液;
(3)、将浆液通过均质机,40-60MPa压力下均质,得均质液;
(4)、将均质液打入酶解罐中,加热至45-55℃为料液;
(5)、料液用10%NaOH调pH7.5-8.5;
(6)、加占蛋白总量1.0-5.0%的复合蛋白酶,搅拌酶解3.0-6.0h;
(7)、料液pH调至5.0-7.0;
(8)、料液升温加热至70-90℃,灭酶10-30min;
(9)、料液经管式离心机分离得水相及油相;
(10)、水相过孔径10-100μm陶瓷膜进一步净化;
(11)、净化液经截留分子量为800-1200Da的有机膜进行分离;
(12)、截留液浓缩至固含量≥25%,经3-6倍体积95%乙醇处理,沉淀经无水乙醇脱水后干燥即得海参肠卵粗多糖;或加入1-10%糊精作为助凝剂,搅拌溶解后喷雾干燥得海参肠卵粗多糖;
(13)、透过液加1-5%β-环糊精,在50℃-70℃搅拌处理15min-60min进行包埋脱腥,脱腥液过颗粒活性炭柱进一步脱腥脱色;
(14)、过柱液用孔径10-100μm陶瓷膜脱炭,得澄清精制液;
(15)、精制液减压蒸发浓缩至固形物30%-40%,得浓缩液;
(16)、将浓缩液喷雾干燥得海参肠卵低聚肽;
(17)、油相经2-5倍体积石油醚反复萃取3-5次,萃取所得有机相经旋转蒸发除尽石油醚获取海参粗卵油。
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