| 鲮鱼皮胶原肽的制备及其抗氧化活性的检测 |
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| 引用本文： | 武翠玲,吴日帮,刘丹,杨兴昊,张姜,黄嘉丰,何海伦.鲮鱼皮胶原肽的制备及其抗氧化活性的检测[J].生物工程学报,2016,32(12):1727-1734. |
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| 作者姓名： | 武翠玲 吴日帮 刘丹 杨兴昊 张姜 黄嘉丰 何海伦 |
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| 作者单位： | 1 中南大学生命科学学院 医学遗传学国家重点实验室，湖南 长沙 410013；2 长治医学院 生物化学教研室，山西 长治 046000,1 中南大学生命科学学院 医学遗传学国家重点实验室，湖南 长沙 410013,1 中南大学生命科学学院 医学遗传学国家重点实验室，湖南 长沙 410013,1 中南大学生命科学学院 医学遗传学国家重点实验室，湖南 长沙 410013,1 中南大学生命科学学院 医学遗传学国家重点实验室，湖南 长沙 410013,1 中南大学生命科学学院 医学遗传学国家重点实验室，湖南 长沙 410013,1 中南大学生命科学学院 医学遗传学国家重点实验室，湖南 长沙 410013 |
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| 基金项目： | 国家自然科学基金 (No. 31370104)，国家星火计划面上项目 (No. 2013GA770009)，长治医学院科研启动基金项目 (No. QDZ201517) 资助。 |
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| 摘 要： | 为了以细菌胞外蛋白酶酶解低值蛋白资源制备抗氧化活性肽以及挖掘新型蛋白酶,采用液体发酵培养的方法对假交替单胞菌Pseudoalteromonas sp.SHK1-2进行发酵产酶,获得胞外蛋白酶粗酶液用于酶解热水法提取的鲮鱼胶原蛋白,通过超滤、sephadex LH-20分子筛层析获得具有DPPH自由基清除能力(35.6%±7%)、氧自由基清除能力(Oxygen radical absorbance capacity,ORAC)及DNA氧化损伤抑制活性的小分子肽,液相色谱质谱联用鉴定该活性肽分子量为776.2 Da,预测氨基酸序列为Thr-Ala-Gly-His-Pro-Gly-Thr-His。ORAC检测验证了人工合成的多肽同样具有抗氧化活性。研究表明细菌胞外蛋白酶在低值资源高值化中具有一定的应用前景,对于新型蛋白酶及其应用的挖掘具有一定的参考意义。
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| 关 键 词： | 抗氧化肽，鲮鱼，胶原蛋白，DPPH，ORAC，DNA氧化损伤 |
| 收稿时间： | 2016/4/21 0:00:00 |
Preparation and antioxidant activity detection of collagen peptide from Cirrhinus molitorella skin |
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| Cuiling Wu,Ribang Wu,Dan Liu,Xinghao Yang,Jiang Zhang,Jiafeng Huang and Hailun He.Preparation and antioxidant activity detection of collagen peptide from Cirrhinus molitorella skin[J].Chinese Journal of Biotechnology,2016,32(12):1727-1734. |
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| Authors: | Cuiling Wu Ribang Wu Dan Liu Xinghao Yang Jiang Zhang Jiafeng Huang and Hailun He |
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| Institution: | 1 State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, Hunan, China; 2 Department of Biochemistry, Changzhi Medical College, Changzhi 046000, Shanxi, China,1 State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, Hunan, China,1 State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, Hunan, China,1 State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, Hunan, China,1 State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, Hunan, China,1 State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, Hunan, China and 1 State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, Hunan, China |
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| Abstract: | We prepared antioxidant peptide through hydrolyzing low-value protein resources with bacterial extracellular proteases and to discovered novel proteases. Crude extracellular protease obtained from Pseudoalteromonas sp. SHK1-2 fermentation was used to hydrolyze collagen extracted from Cirrhinus molitorella skin. Small peptide fraction was isolated from hydrolysate by ultrafiltration and Sephadex LH-20 size exclusion chromatography and showed 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity (35.6%±7%), oxygen radical absorbance capacity and inhibition of DNA oxidation damage. The molecule weight was 776.2 Da, and amino acid sequence was Thr-Ala-Gly-His-Pro- Gly-Thr-His through liquid chromatography mass spectrum. Our findings suggest that peptide obtained from low-value protein of fish waste by hydrolysis with bacterial protease has antioxidant activity. |
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| Keywords: | antioxidant peptode Corrhinus molitorella collageb DPPH OKAC DNA oxidation damage |
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