CN101824457B - 利用凝乳酶制备酪蛋白抗氧化肽的方法 - Google Patents
利用凝乳酶制备酪蛋白抗氧化肽的方法 Download PDFInfo
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Abstract
本发明采用凝乳酶专一性作用于酪蛋白获得具有较高自由基清除率的酪蛋白抗氧化肽,具有抗癌,抗诱导和抗衰老等生物活性,在食品工业、医药等领域得到广泛应用而且作为食品和机体的抗氧化剂,为保健食品的开发提供了一个有前途的基料。研究表明在pH8.0,温度50℃,E/S=2.5%,S%=6%,水解时间为60min时,酪蛋白肽的抗氧化效果即对二苯代苦味酰基最高清除率为90.94%,羟自由基清除率最高为73.33%。
Description
技术领域
本发明属于生物技术领域,利用酶化法生成酪蛋白抗氧化肽,具体的说是利用凝乳酶酶化生成酪蛋白抗氧化肽。
背景技术
研究发现,人类的许多疾病如心肌梗死,动脉硬化,癌症,骨质疏松症和肥胖等都与氧化有关。因此,通过摄入具有抗氧化活性的物质可降低体内的自由基含量,防止氧化反应的发生,提高机体对疾病的抵抗能力。抗氧化剂可分为天然抗氧化剂与合成抗氧化剂,丁基羟基茴香醚和2,6-二叔丁基对甲酚等合成的化学抗氧化剂通常作为食品添加剂用在食品工业上,用于防止食品的氧化变色或者因氧导致的品质劣变,但由于合成抗氧化剂具有一定的毒性和致癌作用,对人体健康存在着潜在的危险,因此更安全的天然抗氧化剂成为近来研究的热点。生物活性肽由于其特定的结构和性质决定其是蛋白质中最有前途的抗氧化物质。
目前制备抗氧化肽底物多源自胶原蛋白、玉米蛋白、大豆蛋白。主要方法为酶解和微生物发酵,常用的酶包括消化道蛋白酶,微生物和植物来源的酶,如胃蛋白酶和胰蛋白酶等。制备抗氧化肽的技术难点为蛋白的限制性水解,特定的氨基酸排列顺序与组成决定了其抗氧化效果。
发明内容
本发明的目的是,以酪蛋白为底料,通过酶化法生成高效高产率的酪蛋白抗氧化肽。
本发明为实现上述目的,所采用的技术方案为:提供一种制备酪蛋白抗氧化肽的制备方法方法:
(1)将酪蛋白溶于蒸馏水中,置电磁搅拌器上,至充分溶解;
(2)1mol氢氧化钠将上述溶液调节pH至8.0
(3)将预热后的酶制剂加入上述溶液。
(4)水解反应30min-90min后100℃灭酶15min,生成酪蛋白水解物。
(5)调节酪蛋白水解物PH值至等电点4.6,离心分离后采用超滤,得到酪蛋白肽。
根据本发明所述的一种制备蛋白抗氧化肽一优选技术方案:所使用的酶制剂为凝乳酶。
根据本发明所述的一种制备蛋白抗氧化肽一优选技术方案:所使用的酶制剂按2%-10%的比例加入。
本发明的有益效果在于:
1、酪蛋白肽浓度为10mg/mL时,二苯代苦味酰基最高清除率为90.94%,酪蛋白肽浓度为8mg/mL时,羟自由基清除率最高为73.33%。
2、抗氧化效果明显并且没有合成抗氧化剂的毒害作用和致癌作用。
3、凝乳酶制备的小分子量抗氧化肽羟自由基清除率明显高于胃蛋白酶,枯草杆菌酶和木瓜蛋白酶制备的抗氧化肽。
4、本发明凝乳酶制备抗氧化肽经济方便,效果显著,并且具有较高的自由基清除能力,易于实现。
具体实施方式
实施例1
称取12.5g酪蛋白溶于200mL蒸馏水中。置电磁搅拌器上,待充分溶解后用1mol氢氧化钠调节pH至8.0。将预热后的酶制剂-凝乳酶按2.5%的比例加入。水解反应时间60min后100℃灭酶15min停止酶解反应,生成酪蛋白水解物。调节酪蛋白水解物pH至4.6去除酶解物中大分子物质,3500r/min离心15min。采用截留分子量3000,1000超滤膜进行分级分离,获得三组抗氧化肽溶液。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明披露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (1)
1.一种制备酪蛋白抗氧化肽的方法,其特征在于:制备酪蛋白肽的工艺为:
(1)将12.5g酪蛋白溶于200mL蒸馏水中,置电磁搅拌器上,至充分溶解;
(2)1mol氢氧化钠将上述溶液调节pH至8.0;
(3)将预热后的酶制剂加入上述溶液;
(4)水解反应30min-90min后100℃灭酶15min,生成酪蛋白水解物;
(5)调节酪蛋白水解物PH值至等电点4.6,离心分离后采用超滤,得到酪蛋白肽;
所使用的酶制剂为凝乳酶;
所使用的酶制剂按2%-10%的比例加入。
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Citations (3)
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EP1188767A1 (en) * | 2000-08-28 | 2002-03-20 | Kraft Foods Holdings, Inc. | Isolated antioxidant peptides from casein and methods for preparing, isolating and identifying antioxidant peptides |
CN1957757A (zh) * | 2006-09-20 | 2007-05-09 | 中国海洋大学 | 一种天然食品抗氧化剂的制备方法 |
CN101008027A (zh) * | 2007-01-16 | 2007-08-01 | 江南大学 | 一种酶法水解酪蛋白同时制备磷酸肽和非磷酸肽的方法 |
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EP1188767A1 (en) * | 2000-08-28 | 2002-03-20 | Kraft Foods Holdings, Inc. | Isolated antioxidant peptides from casein and methods for preparing, isolating and identifying antioxidant peptides |
CN1957757A (zh) * | 2006-09-20 | 2007-05-09 | 中国海洋大学 | 一种天然食品抗氧化剂的制备方法 |
CN101008027A (zh) * | 2007-01-16 | 2007-08-01 | 江南大学 | 一种酶法水解酪蛋白同时制备磷酸肽和非磷酸肽的方法 |
Non-Patent Citations (5)
Title |
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Kunio Suetsuna等.Isolation and characterization of free radical scavenging activities peptides derived from casein.《J. Nutr. Biochem.》.2000,第11卷128-131. * |
Sandrine G. Rival等.Caseins and Casein Hydrolysates. 2. Antioxidative Properties and Relevance to Lipoxygenase Inhibition.《J. Agric. Food Chem.》.2000,第49卷295-302. * |
刘志东等.酪蛋白酶解物的抗氧化活性研究.《天然产物研究与开发》.2008,第20卷(第1期),19-23. * |
吴 丹,赵新淮.高活性酪蛋白抗氧化肽的制备方法研究.《食品科学》.2009,第30卷(第21期),283-287. * |
胡文琴等.酪蛋白酶解物体外抗氧化作用的研究.《食品科学》.2004,第25卷(第4期),158-162. * |
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