碱性蛋白酶水解花粉中肽的鉴定及其与蜂王浆生物活性的比较。
Peptide identification in alcalase hydrolysated pollen and comparison of its bioactivity with royal jelly.
机构信息
Department of Food Science and Technology, Gorgan university of Agricultural Sciences and Natural Resources, Gorgan, Iran.
Department of Food Science and Technology, Gorgan university of Agricultural Sciences and Natural Resources, Gorgan, Iran.
出版信息
Food Res Int. 2019 Feb;116:905-915. doi: 10.1016/j.foodres.2018.09.027. Epub 2018 Sep 11.
Peptides with a similar antioxidant and ACE-inhibitory activity of royal jelly (RJ) generated from Alcalase hydrolysated pollen (AHP) were predicted by Response Surface Methodology (RSM). The model equations were proposed according to the effects of time and enzyme concentration on the antioxidant and ACE-inhibitory activity. The optimum values for Alcalase concentration and hydrolysis time were 1.5% and 4 h, respectively. Later, AHP was prepared and deproteinised to be further analysed using size-exclusion chromatography (SEC). After SEC separation, fractions with the highest activity of ACE-inhibitory, DPPH radical scavenging and ferric-reducing power were purified by RP-HPLC. The highest ACE-inhibitory and DPPH scavenging activity of fractions was found 100% and 66.61%, respectively. The most active fractions were analysed by nano-liquid chromatography and mass spectrometry in tandem (nLC-MS/MS) and a total of 195 peptide sequences were identified. The origins of all peptides were herbal proteins and certain coincidences with previously described bioactive sequences were discussed.
通过响应面法（RSM）预测了从花粉中用碱性蛋白酶水解（AHP）得到的、具有类似蜂王浆（RJ）抗氧化和 ACE 抑制活性的肽。根据时间和酶浓度对抗氧化和 ACE 抑制活性的影响，提出了模型方程。最佳的碱性蛋白酶浓度和水解时间分别为 1.5%和 4 小时。之后，制备 AHP 并进行除蛋白处理，然后使用尺寸排阻色谱（SEC）进一步分析。SEC 分离后，通过反相高效液相色谱（RP-HPLC）对 ACE 抑制活性、DPPH 自由基清除活性和铁还原能力最高的级分进行纯化。发现级分的 ACE 抑制活性和 DPPH 清除活性最高分别为 100%和 66.61%。通过纳升液相色谱和串联质谱（nLC-MS/MS）对最活跃的级分进行分析，共鉴定出 195 条肽序列。所有肽的来源都是植物蛋白，并讨论了与先前描述的生物活性序列的某些巧合。
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