CN117598479A - 一种基于菠萝蜜的复合试剂制作方法 - Google Patents
一种基于菠萝蜜的复合试剂制作方法 Download PDFInfo
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Abstract
本发明涉及菠萝蜜制剂技术领域,具体是指一种基于菠萝蜜的复合试剂制作方法,包括如下步骤:S1:原料的预处理:将菠萝蜜的果肉进行剥离破碎成小颗粒,与冷水混合加热熬制,熬制20分钟后再次加入冷水,熬制15分钟后,手动搅拌得到菠萝蜜匀浆;S2:第一次复合酶可控酶解:通过添加复合酶进行酶解,冷却静置;S3:第二次复合酶可控酶解:增加柠檬酸钠将菠萝蜜匀浆的Ph值增加,添加复合酶进行酶解,冷却至室温,得到酶解混悬液;S4:升温过滤:将酶解混悬液通过90目网筛与110目分别进行过滤得到精细滤液;S5:获得提取物:将精细滤液经过搅拌加热蒸发,得到免疫活性提取物。
Description
技术领域
本发明涉及菠萝蜜制剂技术领域,具体是指一种基于菠萝蜜的复合试剂制作方法。
背景技术
菠萝蜜(学名:Artocarpus heterophyllus Lam),又名木菠萝、树菠萝等,属桑科菠萝蜜属常绿乔木,为海南主要农作物之一。菠萝蜜果肉中含有丰富的糖类、蛋白质、维生素、矿物质等,味道爽口香甜。菠萝蜜种子内部含有大量的碳水化合物和蛋白,其质量约占到菠萝蜜的10%左右,可以炒食或煮食;
目前对菠萝蜜果肉的研究主要集中菠萝蜜水果片、蜜饯、果酒、菠萝蜜种子糕点等的制作工艺研究,而对于菠萝蜜活性物质的研究与开发主要集中在多糖、果胶、黄酮和多酚类物质上。有研究在菠萝蜜中提取到具有免疫活性的多糖。上述物质的提取率低,以及相关具有免疫活性的复合制剂研究较少,菠萝蜜没有得到充分有效利用;
因此,以菠萝蜜为基础,提取制备具有免疫活性的混合物质,进而评价其体内相关免疫活性,对食物来源免疫活性物质的相关研究意义重大,同时为提高菠萝蜜产品附加值,开发辅助提高免疫力产品,奠定了研究基础,并产生较好的经济影响。
发明内容
本发明的目的在于提供一种基于菠萝蜜的复合试剂制作方法,以解决上述背景提到的问题。
本发明所采用的技术方案为:一种基于菠萝蜜的复合试剂制作方法,包括如下步骤:
S1:原料的预处理:将菠萝蜜的果肉进行剥离,然后将菠萝蜜果肉破碎成小颗粒,可通过切碎机来进行破碎,与水温10℃左右的冷水混合后,进行加热熬制,其中菠萝蜜果肉与冷水配比为1-6kg:8-18L,熬制20分钟后再次加入冷水,加入冷水的量为5-8L,二次加水能够使熬制温度降低以便于控温,并且保证不会产生烧糊,熬制15分钟后,手动搅拌冷却至45℃-55℃,得到菠萝蜜匀浆;
S2:第一次复合酶可控酶解:菠萝蜜匀浆初始Ph值为3.5-5.2,然后通过添加复合酶进行酶解,添加时间为5min,在添加的过程中需要不间断搅拌,通过搅拌机在常温下来进行搅拌,搅拌机转速为30-50r/min,搅拌时间设定为10min,便于充分进行融合,然后在85℃-98℃下进行保温灭酶,灭酶结束后,冷却至45℃-55℃静置,以进行第二次酶解;
S3:第二次复合酶可控酶解:通过增加柠檬酸钠将菠萝蜜匀浆的Ph值增加至7.2-8.8,然后通过添加复合酶进行酶解,在添加的过程中需要不间断搅拌,便于充分进行融合,然后在85℃-98℃下进行保温灭酶,冷却至室温,得到酶解混悬液;
S4:升温过滤:将酶解混悬液升温至85℃-95℃,然后通过90目网筛进行过滤,静置冷却至35℃后通过110目得到精细滤液,通过两次过滤,能够将酶解混悬液中的絮凝物去除;
S5:获得提取物:将精细滤液经过搅拌加热蒸发,通过多功能化搅拌加热蒸发仪器来进行搅拌加热蒸发的操作,多功能化搅拌加热蒸发仪器转速为1200-1500r/min,温度为40℃-55℃,以便于将精细滤液中的水分蒸发出来,使精细滤液浓缩至初始容量的1/4-1/10,得到免疫活性提取物。
进一步地,所述免疫活性提取物中的蛋白质含量为整体重量的10.3%-32.5%,多糖含量为35%-80%,淋巴细胞转化值为0.35-0.63,NK细胞活性为55-70%。
进一步地,所述蛋白质中分子量为300-3000Da的多肽质量分数为55-85%。
进一步地,S2中第一次酶解复合酶原料为菠萝蛋白酶、木聚糖酶、辅酶、辅助剂,菠萝蛋白酶、木聚糖酶、辅酶、辅助剂之间的配比为1:5:0.2:0.1混合组成,第一次酶解复合酶的加入质量为菠萝蜜果肉重的0.5-1.5%,酶解时间为2-4h。
进一步地,S3中第二次酶解复合酶原料为溶菌酶、碱性蛋白酶、辅酶、辅助剂,溶菌酶、碱性蛋白酶、辅酶、辅助剂之间的配比为6:2.5:0.2:0.1混合组成,第二次酶解复合酶的加入质量为菠萝蜜果肉重的0.1-0.35%,酶解时间为2-4h。
进一步地,S4中进行过滤之前酶解混悬液的温度处于85℃以上,以保持酶解混悬液内部分子活性,以便于加快过滤速率。
进一步地,所述辅酶为辅酶A或辅酶Q10,所述辅助剂包括保护剂、抗氧化剂。
本发明与现有技术相比优点在于:本发明以菠萝蜜为来源,提取了免疫活性提取物,并将该免疫活性提取物与助剂进行复合,形成复合制剂,利用助剂对免疫活性提取物中的多肽、蛋白质、多糖进行负载,一方面避免活性物质的活性降低,另一方面,提高免疫活性提取物的加工性能,可用于食品、医药和护肤品等方面;所述免疫活性提取物中含有多肽分子量为300-3000Da,蛋白质含量为10.3%-32.5%,多糖含量为35%-80%,且该免疫活性提取物的淋巴细胞转化值为0.35-0.63,NK细胞活性为55-70%;本发明中的提取工艺简单,投入设备较少,产品利用价值高,有助于提高菠萝蜜附加值,可用于工业推广。
具体实施方式
下面将结合本发明实施例,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作出创造性劳动前提下所获得的所有其它实施例,都属于本发明保护的范围。
实施例1:
免疫活性提取物通过以下步骤制成:
S1:原料的预处理:将菠萝蜜的果肉进行剥离,然后将菠萝蜜果肉破碎成小颗粒,可通过切碎机来进行破碎,与水温10℃左右的冷水混合后,进行加热熬制,其中菠萝蜜果肉与冷水配比为2kg:8L,熬制20分钟后再次加入冷水,加入冷水的量为5L,二次加水能够使熬制温度降低以便于控温,并且保证不会产生烧糊,熬制15分钟后,手动搅拌冷却至45℃,得到菠萝蜜匀浆;
S2:第一次复合酶可控酶解:菠萝蜜匀浆初始Ph值为3.7,然后通过添加复合酶进行酶解,添加时间为5min,在添加的过程中需要不间断搅拌,通过搅拌机在常温下来进行搅拌,将搅拌机转速设置为35r/min,搅拌时间设定为10min,便于充分进行融合,然后在85℃下进行保温灭酶,灭酶结束后,冷却至45℃静置,以进行第二次酶解;
S3:第二次复合酶可控酶解:通过增加柠檬酸钠将菠萝蜜匀浆的Ph值增加至7.8,然后通过添加复合酶进行酶解,在添加的过程中需要不间断搅拌,便于充分进行融合,然后在85℃下进行保温灭酶,冷却至室温,得到酶解混悬液;
S4:升温过滤:将酶解混悬液升温至85℃,然后通过90目网筛进行过滤,静置冷却至35℃后通过110目得到精细滤液,通过两次过滤,能够将酶解混悬液中的絮凝物去除;
S5:获得提取物:将精细滤液经过搅拌加热蒸发,通过多功能化搅拌加热蒸发仪器来进行搅拌加热蒸发的操作,将多功能化搅拌加热蒸发仪器转速设置为1200r/min,温度为40℃,以便于将精细滤液中的水分蒸发出来,使精细滤液浓缩至初始容量的1/5,得到免疫活性提取物。
将所得免疫活性提取物进行以下成分测定:
蛋白质含量:采用凯氏定氮法,以干基计;
蛋白质中180-2000Da的多肽含量:按照GB/T22729-2008进行测试;
多糖:采用苯酚硫酸法,且以葡萄糖计,以干基计;
上述成分含量测定结果为:蛋白质含量为13%,其中300-3000Da的多肽占56%,多糖含量为35%;
采用淋巴细胞转化法和NK细胞活性测定法测定其活性。
实施例2:
免疫活性提取物通过以下步骤制成:
S1:原料的预处理:将菠萝蜜的果肉进行剥离,然后将菠萝蜜果肉破碎成小颗粒,可通过切碎机来进行破碎,与水温10℃左右的冷水混合后,进行加热熬制,其中菠萝蜜果肉与冷水配比为3.5kg:12L,熬制20分钟后再次加入冷水,加入冷水的量为6.5L,二次加水能够使熬制温度降低以便于控温,并且保证不会产生烧糊,熬制15分钟后,手动搅拌冷却至50,得到菠萝蜜匀浆;
S2:第一次复合酶可控酶解:菠萝蜜匀浆初始Ph值为4.1,然后通过添加复合酶进行酶解,添加时间为5min,在添加的过程中需要不间断搅拌,通过搅拌机在常温下来进行搅拌,搅拌机转速为35r/min,搅拌时间设定为10min,便于充分进行融合,然后在90℃下进行保温灭酶,灭酶结束后,冷却至50℃静置,以进行第二次酶解;
S3:第二次复合酶可控酶解:通过增加柠檬酸钠将菠萝蜜匀浆的Ph值增加至7.8,然后通过添加复合酶进行酶解,在添加的过程中需要不间断搅拌,便于充分进行融合,然后在90℃下进行保温灭酶,冷却至室温,得到酶解混悬液;
S4:升温过滤:将酶解混悬液升温至90℃,然后通过90目网筛进行过滤,静置冷却至35℃后通过110目得到精细滤液,通过两次过滤,能够将酶解混悬液中的絮凝物去除;
S5:获得提取物:将精细滤液经过搅拌加热蒸发,通过多功能化搅拌加热蒸发仪器来进行搅拌加热蒸发的操作,多功能化搅拌加热蒸发仪器转速为1400r/min,温度为50℃,以便于将精细滤液中的水分蒸发出来,使精细滤液浓缩至初始容量的1/7,得到免疫活性提取物。。
将所得免疫活性提取物进行以下成分测定:
蛋白质含量:采用凯氏定氮法,以干基计;
蛋白质中180-2000Da的多肽含量:按照GB/T22729-2008进行测试;
多糖:采用苯酚硫酸法,且以葡萄糖计,以干基计;
上述成分含量测定结果为:蛋白质含量为25.7%,其中300-3000Da的多肽占66%,多糖含量为55%;
采用淋巴细胞转化法和NK细胞活性测定法测定其活性。
实施例3
S1:原料的预处理:将菠萝蜜的果肉进行剥离,然后将菠萝蜜果肉破碎成小颗粒,可通过切碎机来进行破碎,与水温10℃左右的冷水混合后,进行加热熬制,其中菠萝蜜果肉与冷水配比为6kg:18L,熬制20分钟后再次加入冷水,加入冷水的量为8L,二次加水能够使熬制温度降低以便于控温,并且保证不会产生烧糊,熬制15分钟后,手动搅拌冷却至55℃,得到菠萝蜜匀浆;
S2:第一次复合酶可控酶解:菠萝蜜匀浆初始Ph值为4.3,然后通过添加复合酶进行酶解,添加时间为5min,在添加的过程中需要不间断搅拌,通过搅拌机在常温下来进行搅拌,搅拌机转速为50r/min,搅拌时间设定为10min,便于充分进行融合,然后在98℃下进行保温灭酶,灭酶结束后,冷却至55℃静置,以进行第二次酶解;
S3:第二次复合酶可控酶解:通过增加柠檬酸钠将菠萝蜜匀浆的Ph值增加至8.4,然后通过添加复合酶进行酶解,在添加的过程中需要不间断搅拌,便于充分进行融合,然后在98℃下进行保温灭酶,冷却至室温,得到酶解混悬液;
S4:升温过滤:将酶解混悬液升温至95℃,然后通过90目网筛进行过滤,静置冷却至35℃后通过110目得到精细滤液,通过两次过滤,能够将酶解混悬液中的絮凝物去除;
S5:获得提取物:将精细滤液经过搅拌加热蒸发,通过多功能化搅拌加热蒸发仪器来进行搅拌加热蒸发的操作,多功能化搅拌加热蒸发仪器转速为1500r/min,温度为55℃,以便于将精细滤液中的水分蒸发出来,使精细滤液浓缩至初始容量的1/10,得到免疫活性提取物。
将所得免疫活性提取物进行以下成分测定:
蛋白质含量:采用凯氏定氮法,以干基计;
蛋白质中180-2000Da的多肽含量:按照GB/T22729-2008进行测试;
多糖:采用苯酚硫酸法,且以葡萄糖计,以干基计;
上述成分含量测定结果为:蛋白质含量为30.1%,其中300-3000Da的多肽占76%,多糖含量为69%;
采用淋巴细胞转化法和NK细胞活性测定法测定其活性。
测试方法:选择40只小白鼠,在小白鼠背部相同部位进行刮毛(面积大小相同),进行紫外晒伤试验,其皮肤晒伤程度均在I度晒伤,将40只小白鼠分成5组,一组为对照组,间隔2h对烫伤处进行喷敷生理盐水;其余组分别使用实施例1、实施例2、实施例3获得的复合制剂,将1g复合制剂溶于10g药用生理盐水,间隔2h对烫伤处进行喷敷;喷敷1天后2天后分别观察晒伤处发炎情况;
结果为:
第一天:实施例1无发炎情况、实施例2无发炎情况、实施例3无发炎情况;
第二天:实施例1无发炎情况、实施例2无发炎情况、实施例3无发炎情况;
由此可得知实施例1、实施例2、实施例3获得的复合制剂均具有抗菌作用。
以上实施例仅用以说明本发明的技术方案,而非对其限制;尽管参照前述实施例对本发明进行了详细的说明,本领域的普通技术人员应当理解;其依然可以对前述实施例所记载的技术方案进行修改,或者对其中部分技术特征进行等同替换;而这些修改或者替换,并不使相应技术方案的本质脱离本发明实施例技术方案的精神和范围。
Claims (8)
1.一种基于菠萝蜜的复合试剂制作方法,包括如下步骤:
S1:原料的预处理:将菠萝蜜的果肉进行剥离,然后将菠萝蜜果肉破碎成小颗粒,与冷水混合后,进行加热熬制,熬制20分钟后再次加入冷水,熬制15分钟后,手动搅拌冷却至45℃-55℃,得到菠萝蜜匀浆;
S2:第一次复合酶可控酶解:菠萝蜜匀浆初始Ph值为3.5-5.2,然后通过添加复合酶进行酶解,在添加的过程中需要不间断搅拌,然后在85℃-98℃下进行保温灭酶,灭酶结束后,冷却至45℃-55℃静置,以进行第二次酶解;
S3:第二次复合酶可控酶解:通过增加柠檬酸钠将菠萝蜜匀浆的Ph值增加至7.2-8.8,然后通过添加复合酶进行酶解,再添加的过程中需要不间断搅拌,然后在85℃-98℃下进行保温灭酶,冷却至室温,得到酶解混悬液;
S4:升温过滤:将酶解混悬液升温至85℃-95℃,然后通过90目网筛进行过滤,静置冷却至35℃后通过110目得到精细滤液;
S5:获得提取物:将精细滤液经过搅拌加热蒸发,使精细滤液浓缩至初始容量的1/4-1/10,得到免疫活性提取物。
2.根据权利要求1所述的一种基于菠萝蜜的复合试剂制作方法,其特征在于:所述免疫活性提取物中的蛋白质含量为整体重量的10.3%-32.5%,多糖含量为35%-80%,淋巴细胞转化值为0.35-0.63,NK细胞活性为55-70%。
3.根据权利要求2所述的一种基于菠萝蜜的复合试剂制作方法,其特征在于:所述蛋白质中分子量为300-3000Da的多肽质量分数为55-85%。
4.根据权利要求1所述的一种基于菠萝蜜的复合试剂制作方法,其特征在于:S2中第一次酶解复合酶原料为菠萝蛋白酶、木聚糖酶、辅酶、辅助剂,菠萝蛋白酶、木聚糖酶、辅酶、辅助剂之间的配比为1:5:0.2:0.1混合组成,第一次酶解复合酶的加入质量为菠萝蜜果肉重的0.5-1.5%。
5.根据权利要求1所述的一种基于菠萝蜜的复合试剂制作方法,其特征在于:S3中第二次酶解复合酶原料为溶菌酶、碱性蛋白酶、辅酶、辅助剂,溶菌酶、碱性蛋白酶、辅酶、辅助剂之间的配比为6:2.5:0.2:0.1混合组成,第二次酶解复合酶的加入质量为菠萝蜜果肉重的0.1-0.35%。
6.根据权利要求1所述的一种基于菠萝蜜的复合试剂制作方法,其特征在于:S4中进行过滤之前酶解混悬液的温度处于85℃以上。
7.根据权利要求4所述的一种基于菠萝蜜的复合试剂制作方法,其特征在于:所述辅酶为辅酶A或辅酶Q10,所述辅助剂包括保护剂、抗氧化剂。
8.根据权利要求1-7任意一项所述的复合制剂在食品、医药、护肤品方面均有应用。
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