响应面法优化苦瓜籽蛋白质提取工艺
详细信息Optimization of protein extraction from defatted bitter melon seed using response surface methodology
-
摘要: 以苦瓜籽为原料,采用紫外光谱和响应面法研究盐溶法提取苦瓜籽蛋白质的最佳工艺。以苦瓜籽蛋白质的提取率为指标,考察NaCl浓度、pH、提取时间对苦瓜籽蛋白质提取率的影响。在单因素实验的基础上,通过Box-Benhnken中心组合设计和响应面分析法,确定苦瓜籽蛋白质的最佳提取工艺。结果表明:在料液比为1:6,提取温度为4℃下,NaCl浓度1.3mol/L,pH8.8,提取时间3.6h,苦瓜籽蛋白质的提取率为23.80%,较优化前提高了3.24倍,同时建立了盐提蛋白质的二次多项数学模型,对苦瓜籽蛋白质的提取具有很好的预测作用。此外,通过凯氏定氮法确定苦瓜籽中蛋白质含量为32.13%。Abstract: The optimum extracting conditions of protein from bitter melon seeds by the salt solvent method were studied.The effects of concentration of NaCl, pH and extraction time on the yield of protein were studied based on the extraction rate.According to the single factor experiments, the optimum extract process for bitter melon seed protein was determined through Box-Benhnken central composite design and response surface methodology.The results showed that the optimized extract process for bitter gourd seed protein was that the material/liquid ratio was 1:6 and the temperature was at 4℃, the concentration of NaCl was 1.3mol/L and pH8.8, at a extract time of 3.6h, the extraction rate of bitter gourd seed protein was 23.80%, 3.24 times higher than that before the optimization.In addition, a quadratic equation model for protein extraction was set up and it played an important role in predicting the extraction of protein.Furthermore, the protein content of bitter melon seeds was 32.13% determined by Kjeldahl method.
-
[1] 江苏新医学院.中药大辞典 (上册) [M].上海:上海科技出版社, 1986:1281. [2] Xiong S D, Yu K, Liu X H, et al.Ribosome-inactivating proteins isolated from dietary bitter melon induce apoptosis and inhibit histone deacetylase-1selectively in premalignant and malignant prostate cancer cells[J].International Journal of Cancer, International Journal of Cancer, 2009, 125 (4) :774-782.
[3] Huang L, Adachi T, Shimizu Y, et al.Characterization of lectin isolated from Momordica charantia seed as a B cell activator[J].Immunology letters, 2008, 121 (2) :148-156.
[4] Parkash A, Ng TB, Tso WW.Purification and characterization of charantin, a napin-like ribosome-inactivating peptide from bitter gourd (Momordica charantia) seeds[J].The Journal of Peptide Research:Official Journal of the American Peptide Society, 2002, 59 (5) :197-202.
[5] 蔡马, 于群, 朱新产.苦瓜籽蛋白的PEG/ (NH4) 2SO4双水相分离及抑菌作用的研究[J].天然产物研究与开发, 2011, 23 (4) :612-619. [6] R Horax, N Hettiarachchy, A Kannan, et al.Proximate composition and amino acid and mineral contents of Mormordica charantia L.pericarp and seeds at different maturity stages[J].Food Chemistry, 2010, 122 (4) :1111-1115.
[7] Fang EF, Zhang CZ, Wong JH, et al.The MAP30protein from bitter gourd (Momordica charantia) seeds promotes apoptosis in liver cancer cells in vitro and in vivo[J].Cancer letters, 2012, 324 (1) :66-74.
[8] Fang E F, Zhang C Z, Zhang L, et al.In vitro and in vivo anticarcinogenic effects of RNase MC2, a ribonuclease isolated from dietary bitter gourd, toward human liver cancer cells[J].The International Journal of Biochemistry&Cell Biology, 2012, 44 (8) :1351-1360.
[9] 郎海滨, 糜漫天, 程天民, 等.苦瓜核糖体失活蛋白的纯化及其诱导HL-60细胞凋亡的研究[J].营养学报, 2004, 26 (2) :139-143. [10] Fang E F, Zhang C Z, Ng T B, et al.Momordica Charantia lectin, a type II ribosome inactivating protein, exhibits antitumor activity toward human nasopharyngeal carcinoma cells In Vitro and In Vivo[J].Cancer Prevention Research (Philadelphia, Pa.) , 2012, 5 (1) :109-121.
[11] Jiratchariyakul W, Wiwat C, Vongsakul M, et al.HIV Inhibitor from Thai bitter gourd[J].Planta Medica, 2001, 67 (4) :350-353.
[12] 樊剑鸣, 马建新, 高德富, 等.苦瓜蛋白MAP30对大肠癌细胞株LoVo细胞生长及凋亡的影响[J].郑州大学学报:医学版, 2009, 44 (5) :983-985. [13] Li M, Chen Y, Liu Z, et al.Anti-tumor activity and immunological modification of ribosome-inactivating protein (RIP) from Momordica Charantia by covalent attachment of polyethylene glycol[J].Acta Biochimica et Biophysica Sinica, 2009, 41 (9) :792-799.
[14] Ng T B, Wong C M, Li W W, et al.Isolation and characterization of a galactose binding lectin with insulinomimetic activities.From the seeds of the bitter gourd Momordica charantia (Family Cucurbitaceae) [J].International Journal of Peptide andProtein Research, 1986, 28 (2) :163-172.
[15] Hara S, Makino J, Ikenaka T.Amino acid sequences and disulfide bridges of serine Proteinase inhibitors from bitter gourd (Momordica charantia Linn) seeds[J].Journal of Biochemistry, 1989, 105 (1) :88-92.
[16] Hamato N, Koshiba T, Pham TN, et al.Trypsin and elastase inhibitors from bitter gourd (Momordica charantia Linn) seeds:purification, amino acid sequences, and inhibitory activities of four new inhibitors[J].Journal of Biochemistry, 1995, 117 (2) :432-437.
[17] Huang B, Ng TB, Fong WP, et al.Isolation of a trypsin inhibitor with deletion of N-terminal pentapeptide from the seeds of Momordica cochinchinensis, the Chinese drug mubiezhi[J].The International Journal of Biochemistry&Cell Biology, 1999, 31 (6) :707-715.
[18] Lee-Huang S, Huang PL, Nara PL, et al.MAP30:a new inhibitor of HIV-1infection replication[J].FEBS Letters, 1990, 272 (1-2) :12-18.
[19] Khanna P, Jain SC, Panagariya A, et al.Hypoglycemic activity of polypeptide-p from a plant source[J].Journal of Natural Products, 1981, 44 (6) :648-655.
[20] Horax R, Hettiarachchy N, Over K, et al.Extraction, Fractionation and Characterization of Bitter Melon Seed Proteins[J].Journal of Agricultural and Food Chemistry, 2010, 58 (3) :1892-1897.
[21] 王镜岩.生物化学 (上册) [M].北京:高等教育出版社, 2002:305. [22] 周瑞宝.植物蛋白功能原理与工艺[M].北京:化学工业出版社, 2007:30. [23] 张建社.蛋白质分离与纯化技术[M].北京:军事医学科学出版社, 2009:93.
计量
- 文章访问数: 102
- HTML全文浏览量: 11
- PDF下载量: 814
下载:
