酶法破壁提取蜂花粉功能成分的工艺优化及抗氧化活性分析

Process optimization of enzymatic wall-disruption and extraction of functional components in bee pollen and analysis of antioxidant activities

中文关键词:  蜂花粉，破壁，酶解，碱处理，酚类物质

英文关键词:bee pollen  wall-disruption  enzymatic  alkali treatment  phenols

中文摘要:

      目的 探索酶解结合超声和碱解从而高效破壁和提取蜂花粉中的活性成分。方法 采用扫描电镜对蜂花粉微观结构进行分析, 并使用高效液相色谱法分析酚类物质的组成和含量变化。结果 经12.5%碱性蛋白酶酶解1 h后, 蜂花粉酶解上清液的总酚含量、总黄酮含量和2,2’-联氮-二(3-乙基-苯并噻唑啉-6-磺酸)二铵盐[2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) ammonium salt, ABTS]阳离子自由基清除能力最高, 分别为(14.14±0.23) mg GAE/(g·dw), (2.34±0.04) mg RE/(g·dw)和(167.06±1.39) μmol TE/(g·dw)。将酶解辅以超声协同0.3 mol/L NaOH处理后, 总酚、总黄酮、ABTS阳离子自由基清除能力及铁离子还原能力相比未处理花粉分别提高了191%、39%、69%和162%。扫描电镜结果显示, 碱处理酶解后花粉细胞壁破碎程度显著增加, 花粉内含物质从裂口处向外溢出, 与理化指标趋势一致。最佳工艺条件为0.3 mol/L NaOH超声-碱处理1 h, 调节pH至7, 12.5%的碱性蛋白酶于50℃下酶解1 h。结论 超声碱处理可辅助酶解实现蜂花粉的高效破壁, 同时小肽、多酚、黄酮等营养成分也进一步溶出, 促进了蜂花粉的高值化利用。

英文摘要:

      Objective To explore the potential combination of enzymatic hydrolysis and treatments with ultrasonication or alkaline hydrolysis with the purpose of achieving the effective wall-disruption and extraction of nutrients in bee pollen. Methods Scanning electron microscopy was used to observe the microstructure of bee pollen, and high performance liquid chromatography was used to achieve the qualitative and quantitative analysis of phenols. Results The antioxidant values such as total phenols, total flavonoids, and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) ammonium salt (ABTS) radical scavenging ability of bee pollen enzymatic supernatant turned to be the highest in the treatment with 12.5% alkaline protease for 1 h, with the specific values reaching (14.14±0.23) mg GAE/(g·dw), (2.34±0.04) mg RE/(g·dw) and (167.06±1.39) μmol TE/(g·dw), respectively. Total phenols, total flavonoids, ABTS free radical scavenging ability and iron ion reduction ability increased by 191%, 39%, 69% and 162% respectively compared with untreated pollen after enzymatic hydrolysis combined with ultrasound and 0.3 mol/L NaOH treatment. The results of scanning electron microscopy revealed that the wall-disruption of bee pollen was further intensified after the treatment with enzymatic hydrolysis-ultrasonication- alkaline hydrolysis, and the pollen content were spilled outward from the clefts, which was consistent with the trends of physicochemical indexes. The optimal process conditions were 0.3 mol/L NaOH ultrasonic-alkali treatment for 1 h, adjusting the pH to 7, and enzymatic hydrolysis with 12.5% alkaline protease at 50℃ for 1 hour. Conclusion Ultrasonic alkali treatment can assist enzymatic hydrolysis to break the wall of bee pollen, and nutrients such as small peptides, polyphenols and flavonoids are further dissolved, which promotes the high-value utilization of bee pollen.