Faculty of Food Technology and Biotechnology, University of Zagreb, 10000 Zagreb, Croatia.
Department of Biology, Faculty of Science, University of Zagreb, 10000 Zagreb, Croatia.
Ultrason Sonochem. 2023 Oct;99:106535. doi: 10.1016/j.ultsonch.2023.106535. Epub 2023 Jul 27.
Ultrasound (US) and high voltage electric discharge (HVED) with water as a green solvent represent promising novel non-thermal techniques for protein extraction from sugar beet (Beta vulgaris subsp. vulgaris var. altissima) leaves. Compared to HVED, US proved to be a better alternative method for total soluble protein extraction with the aim of obtaining high yield of ribulose-1,5-bisphosphate carboxylase-oxygenase enzyme (RuBisCO). Regardless of the solvent temperature, the highest protein yields were observed at 100% amplitude and 9 min treatment time (84.60 ± 3.98 mg/g with cold and 96.75 ± 4.30 mg/g with room temperature deionized water). US treatments at 75% amplitude and 9 min treatment time showed the highest abundance of RuBisCO obtained by immunoblotting assay. The highest protein yields recorded among HVED-treated samples were observed at a voltage of 20 kV and a treatment time of 3 min, disregarding the used gas (33.33 ± 1.06 mg/g with argon and 34.89 ± 1.59 mg/g with nitrogen as injected gas), while the highest abundance of the RuBisCO among HVED-treated samples was noticed at 25 kV voltage and 3 min treatment time. By optimizing the US and HVED parameters, it is possible to affect the solubility and improve the isolation of RuBisCO, which could then be purified and implemented into new or already existing functional products.
超声（US）和高压电放电（HVED）与水作为绿色溶剂结合，代表了从糖甜菜（Beta vulgaris subsp. vulgaris var. altissima）叶片中提取蛋白质的有前途的新型非热技术。与 HVED 相比，US 被证明是一种更好的替代方法，用于提取总可溶性蛋白质，目的是获得高产量的核酮糖-1,5-二磷酸羧化酶-加氧酶（RuBisCO）。无论溶剂温度如何，在 100%幅度和 9 分钟处理时间下（冷水中为 84.60 ± 3.98mg/g，室温去离子水中为 96.75 ± 4.30mg/g）观察到最高的蛋白质产量。在 75%幅度和 9 分钟处理时间的 US 处理下，通过免疫印迹分析获得的 RuBisCO 丰度最高。在 HVED 处理的样品中，记录到的最高蛋白质产量出现在电压为 20 kV 和处理时间为 3 分钟的情况下，不考虑使用的气体（氩气为 33.33 ± 1.06mg/g，氮气为 34.89 ± 1.59mg/g），而在 HVED 处理的样品中 RuBisCO 的丰度最高出现在 25 kV 电压和 3 分钟处理时间。通过优化 US 和 HVED 参数，可以影响 RuBisCO 的溶解度并改善其分离，然后可以对其进行纯化并应用于新的或现有的功能性产品中。
