Ricarte Gabriella N, Coelho Maria Alice Z, Marrucho Isabel M, Ribeiro Bernardo Dias
Escola de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-598 Brazil.
Centro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, 1049-001 Lisboa, Portugal.
3 Biotech. 2020 Sep;10(9):405. doi: 10.1007/s13205-020-02393-0. Epub 2020 Aug 25.
The aim of this work is to develop an optimized enzymatic assisted extraction methodology to extract carotenoids and phenolic compounds from sunflower wastes (petals and florets) using natural hydrophobic green solvents. Several natural green hydrophobic solvents were used as well as natural hydrophobic eutectic solvents composed of d,l-menthol and different acids, with different hydrophobicity. The multi-enzyme complex Viscozyme was used to disrupt the cell wall of petals and disc florets. The extracted carotenoids content into the hydrophobic phase was quantified using UV-Vis spectrophotometry and the carotenoids profile was studied using high-performance liquid and thin layer chromatography. The amount of total sugars in the aqueous phase was also analyzed using the dinitrosalicylic acid (DNS) method to infer about the enzymatic action in cell wall. Phenolic compounds also in the aqueous phase were analyzed by Folin Denis method. The eutectic solvent d,l-menthol:d,l-lactic acid (M:HLac) (1:2) was the best solvent for extraction of carotenoids from sunflower wastes, with 147 ppm of carotenoids extracted, in comparison to 115 ppm obtained with the standard solvent, -hexane. In what concerns phenolic compounds, M:HLac was again better than the standard solvent. The use of the multi-enzyme complex Viscozyme had different responses, depending on the solvent tested. For the green solvent M:HLac, the enzyme improved the carotenoids extraction, achieving 335 ppm of carotenoids in the extract. The role of enzyme, solvent, water and sunflower quantity in the carotenoid extraction was evaluated and optimized through a central composite rotatable design (CCRD), using the M:HLac as solvent. According to the analysis of CCRD, the most efficient extractions were carried out using more solvent and less raw material, whose best result reached 1449 mg carotenoids/100 g biomass ppm of carotenoids. This work emphasizes the possibility of developing more sustainable enzyme-assisted separation processes, through the substitution of toxic solvents with natural, environmentally friendly, solvents.
这项工作的目的是开发一种优化的酶辅助提取方法，使用天然疏水性绿色溶剂从向日葵废料（花瓣和小花）中提取类胡萝卜素和酚类化合物。使用了几种天然绿色疏水性溶剂以及由d，l-薄荷醇和不同酸组成的具有不同疏水性的天然疏水性低共熔溶剂。使用多酶复合物Viscozyme破坏花瓣和盘状小花的细胞壁。使用紫外可见分光光度法定量疏水性相中提取的类胡萝卜素含量，并使用高效液相色谱和薄层色谱研究类胡萝卜素谱。还使用二硝基水杨酸（DNS）方法分析水相中的总糖量，以推断细胞壁中的酶促作用。水相中的酚类化合物通过Folin Denis方法进行分析。低共熔溶剂d，l-薄荷醇：d，l-乳酸（M：HLac）（1：2）是从向日葵废料中提取类胡萝卜素的最佳溶剂，提取出147 ppm的类胡萝卜素，相比之下，标准溶剂正己烷提取的为115 ppm。在酚类化合物方面，M：HLac再次优于标准溶剂。根据所测试的溶剂不同，多酶复合物Viscozyme的使用有不同的反应。对于绿色溶剂M：HLac，该酶改善了类胡萝卜素的提取，提取物中类胡萝卜素含量达到335 ppm。以M：HLac为溶剂，通过中心复合旋转设计（CCRD）评估并优化了酶、溶剂、水和向日葵用量在类胡萝卜素提取中的作用。根据CCRD分析，使用更多溶剂和更少原料进行的提取效率最高，其最佳结果达到1449 mg类胡萝卜素/100 g生物质ppm类胡萝卜素。这项工作强调了通过用天然、环境友好的溶剂替代有毒溶剂来开发更可持续的酶辅助分离工艺的可能性。
