Faculty of Chemistry, Al. I. Cuza University of Iasi, 11, Carol I Boulevard, 700506 Iasi, Romania.
Department of Chemistry & Biomolecular Science, Clarkson University, 8 Clarkson Avenue, Potsdam, NY 13699, USA.
Biomolecules. 2021 Dec 6;11(12):1838. doi: 10.3390/biom11121838.
Zein is a type of prolamin storage protein that has a variety of biomedical and industrial applications. Due to the considerable genetic variability and polyploidity of the starting material, as well as the extraction methods used, the characterization of the protein composition of zein requires a combination of different analytical processes. Therefore, we combined modern analytical methods such as mass spectrometry (MS), Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), atomic force microscopy (AFM), or Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) for a better characterization of the extracted zein. In this study, we present an enhanced eco-friendly extraction method, including grinding and sieving corn seeds, for prolamins proteins using an ultrasonic extraction methodology. The use of an ultrasonic homogenizer, 65% ethanol extraction buffer, and 710 µm maize granulation yielded the highest protein extraction from all experimental conditions we employed. An SDS PAGE analysis of the extracted zein protein mainly revealed two intense bands of approximatively 20 and 23 kDa, suggesting that the extracted zein was mostly α-zein monomer. Additionally, MS analysis revealed as a main component the α-zein PMS2 (Uniprot accession no. P24450) type protein in the maize flour extract. Moreover, AFM studies show that extracting zein with a 65% ethanol and a 710 µm granulation yields a homogeneous content that could allow these proteins to be employed in future medical applications. This research leads to a better understanding of zeins content critical for developing new applications of zein in food and pharmaceutical industries, such as biocompatible medical vehicles based on polyplexes complex nanoparticles of zein with antimicrobial or drug delivery properties.
玉米醇溶蛋白是一种醇溶蛋白储存蛋白，具有多种生物医学和工业应用。由于起始材料的遗传变异性和多倍性以及所使用的提取方法，玉米醇溶蛋白的蛋白质组成的特性需要结合不同的分析过程。因此，我们结合了现代分析方法，如质谱（MS）、十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）、原子力显微镜（AFM）或傅里叶变换红外光谱-衰减全反射（FTIR-ATR），以更好地表征提取的玉米醇溶蛋白。在这项研究中，我们提出了一种增强的环保提取方法，包括研磨和筛分玉米种子，以及使用超声波提取方法提取醇溶蛋白。使用超声波匀浆器、65%乙醇提取缓冲液和 710 µm 玉米粒可从我们采用的所有实验条件中获得最高的蛋白质提取率。提取的玉米醇溶蛋白的 SDS-PAGE 分析主要显示出两条大约 20 和 23 kDa 的强烈条带，表明提取的玉米醇溶蛋白主要是α-醇溶蛋白单体。此外，MS 分析表明，在玉米粉提取物中，主要成分是α-醇溶蛋白 PMS2（Uniprot 登记号 P24450）类型蛋白。此外，AFM 研究表明，用 65%乙醇和 710 µm 粒度提取醇溶蛋白可得到均匀的含量，这可能使这些蛋白质能够用于未来的医学应用。这项研究有助于更好地了解玉米醇溶蛋白的含量，这对于开发玉米醇溶蛋白在食品和制药行业的新应用至关重要，例如基于具有抗菌或药物递送性能的醇溶蛋白与多聚物复合纳米颗粒的生物相容医学载体。
