Departamento de Química Analítica, Química Física e Ingeniería Química, Instituto de Investigación Química "Andrés M. del Río", Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871, Alcalá de Henares (Madrid), Spain.
Departamento de Química Analítica, Química Física e Ingeniería Química, Instituto de Investigación Química "Andrés M. del Río", Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871, Alcalá de Henares (Madrid), Spain.
J Chromatogr A. 2019 Jun 7;1594:82-92. doi: 10.1016/j.chroma.2019.02.018. Epub 2019 Feb 7.
This work proposes a method to separate proteins and polyphenols in a food byproduct with high bioactive properties and demonstrate by high performance liquid chromatography tandem mass spectrometry its efficiency. Bioactive substances were extracted using high intensity focused ultrasounds. Resulting extract (SR) was submitted to a step for the purification of proteins and to obtain a protein isolate (PI). Both extracts (SR and PI) were digested using two different enzymes (alcalase and thermolysin). Antioxidant, hypocholesterolemic, and antihypertensive properties of hydrolysates were explored. High antioxidant capacity, mainly due to the presence of polyphenols, was observed in SR and its hydrolysates. Hydrolysates obtained from PI using the alcalase enzyme showed simultaneously a high capacity to inhibit cholesterol esterase and to reduce micellar cholesterol solubility. Hydrolysate obtained from PI using the thermolysin enzyme showed a high antihypertensive capacity. Peptides and polyphenols in hydrolysates were identified by RP-HPLC-ESI-Q-TOF. Hydrolysates obtained from PI showed a high amount of peptides and a negligible amount of polyphenols while polyphenols were mainly present in hydrolysates from SR.
本工作提出了一种从具有高生物活性特性的食品副产物中分离蛋白质和多酚的方法，并通过高效液相色谱串联质谱法证明了其效率。使用高强度聚焦超声提取生物活性物质。所得提取物（SR）经过蛋白质纯化步骤以获得蛋白质分离物（PI）。将两种提取物（SR 和 PI）分别用两种不同的酶（碱性蛋白酶和胰酶）进行酶解。研究了水解产物的抗氧化、降胆固醇和降血压特性。SR 及其水解产物具有高抗氧化能力，主要归因于多酚的存在。用碱性蛋白酶水解 PI 得到的水解产物同时具有高胆固醇酯酶抑制能力和降低胶束胆固醇溶解度的能力。用胰酶酶解 PI 得到的水解产物具有高的降压能力。通过反相高效液相色谱-电喷雾-四极杆-飞行时间质谱（RP-HPLC-ESI-Q-TOF）鉴定了水解产物中的肽和多酚。PI 来源的水解产物中含有大量肽和极少量多酚，而 SR 来源的水解产物中主要含有多酚。
