CN107691792A - 一种鲎肽膏的制备方法及其产物和应用 - Google Patents
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Abstract
本发明涉及一种鲎肽膏的制备方法及其产物和应用,其制备方法包括如下步骤:1)采集鲎的肉液和籽,通过胶体磨磨浆,得到细浆液;2)在细浆液中加入动物蛋白酶和风味酶进行水解,待水解结束后升温杀酶,取得酶解液;3)将酶解液真空浓缩,得到鲎肽膏。该制备方法得到的鲎肽膏能够作为诱食剂,增加实验鼠的摄食欲望,保证实验鼠的摄食量,提高动物模型的造模质量。
Description
技术领域
本发明涉及实验动物饲料添加剂的制备领域,具体涉及一种鲎肽膏的制备方法及其产物和应用。
背景技术
实验鼠主要包括大鼠和小鼠,实验鼠在毒理、药理、药效等生命科学领域的动物实验中发挥着非常重要的作用。用鼠做实验有很多优点,首先实验鼠与人类的DNA差异很小,有很近的亲缘关系,对后期的临床实验有很重要的指导意义;其次实验鼠与人有很多共患的疾病,便于造模;最后实验鼠繁殖能力强,性成熟快,繁殖数量多,来源充足,而且价格便宜。
在大、小鼠的饲养过程中,一般饲喂全价营养颗粒饲料,饲料中含一定比例的粗纤维,使成型后饲料具一定的硬度,以便实验鼠磨牙。实验鼠饲养采用群养,摄食采用自由摄食,因此为保证实验数据的稳定和可靠,必须让每只鼠均有充足饮食,所以饲喂的饲料就必须满足实验鼠的食用喜好和营养。
一般实验情况下全价营养颗粒饲料可以保证上述饲喂要求,但糖尿病鼠、冠心病鼠和高血脂鼠等疾病模型动物需饲喂的专用饲料常常会使实验鼠产生厌食,减少饲料的摄入量,进而影响实验的稳定性和可靠性。
发明内容
本发明的目的在于针对现有技术的不足,提供一种鲎肽膏的制备方法及其产物和应用,该制备方法得到的鲎肽膏能够作为诱食剂,增加实验鼠的摄食欲望,保证实验鼠的摄食量,提高动物模型的造模质量。
本发明针对上述的技术问题所提供的技术方案为:
一种鲎肽膏的制备方法,包括如下步骤:
1)采集鲎的肉液和籽,通过胶体磨磨浆,得到细浆液;
2)在细浆液中加入动物蛋白酶和风味酶进行水解,待水解结束后升温杀酶,取得酶解液;
3)将酶解液真空浓缩,得到鲎肽膏。
上述技术方案中,采用鲎的肉液和籽作为原料,由于鲎壳具有很高的营养和药用价值,常用于生产甲壳素,而肉液和籽往往被作为废物。采用去壳后的肉液和籽进行废物利用,可降低废弃物的处理成本,减少对环境的污染。
此外,细浆液通过动物蛋白酶和风味酶进行水解,能够对蛋白质不同部位肽键进行内切和外切,形成分子量较小的肽,同时游离出氨基酸,增加了生物学活性,释放出对实验鼠具有诱食作用的肽基团,使得制备成的鲎肽膏,具有蛋白高、脂肪低的特点,对实验鼠具有极强的诱食功能和营养功能。
优选的,所述步骤1)中采集鲎的肉液和籽具体为:将清洗干净的鲎铡成块状,送入采肉机中进行采集肉液和籽。进一步优选,所述块状的尺寸为5cm×5cm块状。
优选的,所述步骤1)中胶体磨磨浆时加入水,使得细浆液的体积百分比浓度为16-24%。加水量根据取得的肉液和籽的含水量决定。
优选的,所述步骤1)中胶体磨磨浆细度120-180目。进一步优选为150目。
优选的,所述步骤2)中水解前,细浆液采用磷酸调整pH至6.8-7.2。
优选的,所述步骤2)中水解温度为53-56℃,水解时间为2-4h。
优选的,所述步骤2)中细浆液与动物蛋白酶的投料质量比为18000-22000:1;所述细浆液与风味酶的投料质量比为38000-42000:1。进一步优选,所述步骤2)中细浆液与动物蛋白酶的投料质量比为20000:1;所述细浆液与风味酶的投料质量比为40000:1。
优选的,所述步骤2)中杀酶温度为75-90℃,时间为8-12min。
优选的,所述步骤3)中真空浓缩至含水率小于40%。进一步优选,酶解液真空浓缩后可以密封包装。
本发明还提供一种如上述的制备方法制备得到的鲎肽膏。
本发明还提供一种如上述的鲎肽膏作为实验鼠诱食剂的应用。
优选的,所述实验鼠为糖尿病、冠心病或高血脂等疾病的模型鼠。由于这些特殊的疾病模型鼠饲喂专用饲料时,常常会使实验鼠产生厌食,减少饲料的摄入量,进而影响实验的稳定性和可靠性。而鲎肽膏作为实验鼠诱食剂时,能够有效得增加实验鼠的摄食欲望,保证实验鼠的摄食量,提高动物模型的造模质量。
同现有技术相比,本发明的有益效果体现在:
(1)本发明所提供的制备方法,采用去壳后的肉液和籽进行废物利用,可降低废弃物的处理成本,减少对环境的污染。
(2)本发明所提供的制备方法制备得到的鲎肽膏,具有蛋白高、脂肪低的特点,对实验鼠具有极强的诱食功能和营养功能。
(3)本发明所提供的制备方法工艺简单、合理、实施方便,并易规模化生产。
附图说明
图1为鲎肽膏对SD大鼠摄食量的影响曲线图;
图2为鲎肽膏对SD大鼠体重增长的影响曲线图;
图3为鲎肽膏对SD大鼠总胆固醇生化指标的比较图。
具体实施方式
以下实施例用来解释说明本发明,而不是对本发明进行限制,在本发明的精神和权利要求的保护范围内,对本发明做出的任何修改和改变都落入本发明的保护范围。
实施例1:制备鲎肽膏
(1)将清洗干净的5.4公斤鲎铡成5cm×5cm块状,送入采肉机中进行采集肉和籽,取其中的3.5公斤的肉和籽;
(2)将3.5公斤的肉和籽,加入胶体磨加少量水,得到细度为150目浓度为20%的细浆液;
(3)将细浆液采用磷酸调整pH至6.8-7.2,加入180mg动物蛋白酶和90mg风味蛋白酶,控制水解温度为53℃,时间为4h。
(4)水解结束将温度加热至80℃进行杀酶,时间为10min;
(5)将酶解液真空浓缩至含水率20%,得到1公斤鲎肽膏产品,密封包装。
实施例2:制备鲎肽膏
(1)将清洗干净的4.2公斤鲎铡成5cm×5cm块状,送入采肉机中进行采集肉和籽,取其中的3.2公斤的肉和籽;
(2)将3.2公斤的肉和籽,加入胶体磨加少量水,得到细度为180目浓度为16%的细浆液;
(3)将细浆液采用磷酸调整pH至6.8-7.2。加入160mg动物蛋白酶和80mg风味蛋白酶,控制水解温度为55℃,时间为4h。
(4)水解结束将温度加热至85℃进行杀酶,时间为12min;
(5)将酶解液真空浓缩至含水率35%,得到1公斤鲎肽膏产品,密封包装。
实施例3:制备鲎肽膏
(1)将清洗干净的4.6公斤鲎铡成5cm×5cm块状,送入采肉机中进行采集肉和籽,取其中的3.3公斤的肉和籽;
(2)将3.3公斤的肉和籽,加入胶体磨加少量水,得到细度为130目浓度为24%的细浆液;
(3)将细浆液采用磷酸调整pH至6.8-7.2。加入170mg动物蛋白酶和85mg风味蛋白酶,控制水解温度为56℃,时间为4h。
(4)水解结束将温度加热至90℃进行杀酶,时间为8min;
(5)将酶解液真空浓缩至含水率10%,得到1公斤鲎肽膏产品,密封包装。
应用例:鲎肽膏刺激SD大鼠对冠心病模型饲料的摄食效果
一、实验动物
36只健康雄性(SPF级),体重180-200g,购自上海斯莱克实验动物有限责任公司。
冠心病模型大鼠饲料配方为2%胆固醇、10%的猪油、0.2%丙基硫氧嘧啶、0.5%的胆酸钠、87.3%的基础饲料。
含诱食剂的冠心病模型大鼠饲料配制,其中的鲎肽膏由实施例1制备得到:
(1)含1%鲎肽膏:1%鲎肽膏、2%胆固醇、10%的猪油、0.2%丙基硫氧嘧啶、0.5%的胆酸钠、86.3%的基础饲料。
(2)含2%鲎肽膏:2%鲎肽膏、2%胆固醇、10%的猪油、0.2%丙基硫氧嘧啶、0.5%的胆酸钠、85.3%的基础饲料。
(3)含5%鲎肽膏:5%鲎肽膏、2%胆固醇、10%的猪油、0.2%丙基硫氧嘧啶、0.5%的胆酸钠、82.3%的基础饲料。
(4)含10%鲎肽膏:10%鲎肽膏、2%胆固醇、10%的猪油、0.2%丙基硫氧嘧啶、0.5%的胆酸钠、77.3%的基础饲料。
上述配方饲料制备成颗粒饲料后进行钴60辐照。
二、实验方法
取健康SD大鼠36只,随机分成6组,每组6只,分别为组Ⅰ(对照组),饲喂正常基础饲料;组Ⅱ(模型饲料对照组),饲喂模型饲料;组Ⅲ(1%鲎肽膏),饲喂含1%鲎肽膏的模型饲料;组Ⅳ(2%鲎肽膏),饲喂含2%鲎肽膏的模型饲料;组Ⅴ(5%鲎肽膏),饲喂含5%鲎肽膏的模型饲料;组Ⅵ(10%鲎肽膏),饲喂含10%鲎肽膏的模型饲料。连续饲喂30天观察各组实验动物的摄食情况、体重增长情况、生化指标改变情况。
三、实验结果
1.各组SD大鼠的摄食情况:
如图1所示,各组SD大鼠在实验期间每隔3天进行一次摄食量的检测,结果饲喂模型饲料的实验鼠摄食量明显下降,而加入诱食剂后可以明显提高实验鼠的摄食量,且诱食剂的含量与摄食量成正相关。在实验前9天饲喂含10%鲎肽膏的模型饲料的摄食量甚至高于对照组摄食量。结果说明本发明所述鲎肽膏可以有效增加SD大鼠的摄食。
2.各组SD大鼠的体重增长情况:
如图2所示,各组SD大鼠在实验期间每隔3天进行一次体重称量,结果饲喂模型饲料的实验鼠的体重增长速度明显低于对照组,而加入诱食剂后在实验前期可以明显提高SD大鼠的体重增长速度。
3.诱食剂对总胆固醇生化指标的影响:
各组SD大鼠在实验分别在实验第16天、31天各组随机取3只,用1.0%的戊巴比妥钠腹腔注射麻醉后,腹主动脉采血并分离血清,用全自动生化分析仪检测总胆固醇的含量。
如图3所示,在实验第16天,饲喂含鲎肽膏的模型饲料的SD大鼠血清中总胆固醇的含量明显升高(与饲喂模型饲料的SD大鼠比较,P<0.01),且总胆固醇的含量与鲎肽膏的掺入量成正相关,特别是饲喂含5%和10%鲎肽膏的模型饲料后,血清中总胆固醇在非常短的时间内即可达到非常高的水平。
同样在实验第31天,饲喂含鲎肽膏的模型饲料的SD大鼠血清中总胆固醇的含量也出现一定的升高(与饲喂模型饲料的SD大鼠比较),但升高幅度较第16天明显下降。说明利用鲎肽膏作为诱食剂制备冠心病或高血脂动物模型可以减少造模时间。
Claims (10)
1.一种鲎肽膏的制备方法,其特征在于,包括如下步骤:
1)采集鲎的肉液和籽,通过胶体磨磨浆,得到细浆液;
2)在细浆液中加入动物蛋白酶和风味酶进行水解,待水解结束后升温杀酶,取得酶解液;
3)将酶解液真空浓缩,得到鲎肽膏。
2.根据权利要求1所述的鲎肽膏的制备方法,其特征在于,所述步骤1)中采集鲎的肉液和籽具体为:将清洗干净的鲎铡成块状,送入采肉机中进行采集肉液和籽。
3.根据权利要求1所述的鲎肽膏的制备方法,其特征在于,所述步骤1)中胶体磨磨浆细度120-180目。
4.根据权利要求1所述的鲎肽膏的制备方法,其特征在于,所述步骤2)中水解前,细浆液采用磷酸调整pH至6.8-7.2。
5.根据权利要求1所述的鲎肽膏的制备方法,其特征在于,所述步骤2)中水解温度为53-56℃,水解时间为2-4h。
6.根据权利要求1所述的鲎肽膏的制备方法,其特征在于,所述步骤2)中细浆液与动物蛋白酶的投料质量比为18000-22000:1;所述细浆液与风味酶的投料质量比为38000-42000:1。
7.根据权利要求1所述的鲎肽膏的制备方法,其特征在于,所述步骤2)中杀酶温度为75-90℃,时间为8-12min。
8.根据权利要求1所述的鲎肽膏的制备方法,其特征在于,所述步骤3)中真空浓缩至含水率小于40%。
9.一种如权利要求1~8任一所述的制备方法制备得到的鲎肽膏。
10.一种如权利要求9所述的鲎肽膏作为实验鼠诱食剂的应用。
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