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对流干燥木瓜和菠萝残渣中内肽酶的提取与鉴定:一种应用于更大规模的方法学途径。

Extraction and identification of endopeptidases in convection dried papaya and pineapple residues: A methodological approach for application to higher scale.

作者信息

Rojas Luisa Fernanda, Cortés Carolina Flórez, Zapata Paola, Jiménez Claudio

机构信息

Universidad de Antioquia, Grupo de Biotransformación - Escuela de Microbiología, Calle 70 No 52-21, A.A 1226 Medellín, Colombia.

Instituto de Ciencia y Tecnología Alimentaria, INTAL, Carrera. 50 G # 12 Sur 91, Itagüí, Colombia.

出版信息

Waste Manag. 2018 Aug;78:58-68. doi: 10.1016/j.wasman.2018.05.020. Epub 2018 May 28.

DOI:10.1016/j.wasman.2018.05.020
PMID:32559948
Abstract

The use of agro-industrial waste for application in the obtention of products with high added value has become a trend in recent years, especially in tropical countries whose main economic sector is agricultural exports. In the present study, an applicable method to food industry of extracting proteolytic enzymes from dryed papaya and pineapple residues by convection was developed. Different to other scientific reports the heat treatment at 40 °C of waste residues, to reach 20% moisture, allowed an increase in total soluble protein content and did not alter the proteolytic activity of the extracts when phosphate buffer pH 7.0 was used as solvent. In the residues evaluated as candidates for the extraction of endopeptidases, we observed that green dried papaya peel and dried pineapple core, had higher activity values (914.34 ± 25.47 U/mg and 2152.36 ± 75.99 U/mg, respectively). These results, combined with one-dimensional electrophoresis and protein identification methods by MALDI TOF-TOF, showed the presence of signal peptides characteristic of papain, bromelain and other endopeptidases previously reported in extracts of fresh papaya and pineapple residues. These findings show that the drying of the residues by convection does not alter neither the activity nor the structure of the proteolytic enzymes. Finally, it is confirmed that the use of 20% ammonium sulfate as a precipitating agent allows to reach an efficiency of 74% in different work scales the use of purification and identification protocols in a more adaptable way, making them the most promising waste in Colombia, due to its potential for the production of bromelain on a larger scale.

摘要

近年来,利用农工业废弃物获取高附加值产品已成为一种趋势,特别是在主要经济部门为农产品出口的热带国家。在本研究中,开发了一种适用于食品工业的方法,通过对流从干燥的木瓜和菠萝残渣中提取蛋白水解酶。与其他科学报告不同的是,将废渣在40°C下进行热处理以达到20%的含水量,当使用pH 7.0的磷酸盐缓冲液作为溶剂时,可使总可溶性蛋白含量增加,且不会改变提取物的蛋白水解活性。在被评估为内肽酶提取候选物的残渣中,我们观察到绿色干燥木瓜皮和干燥菠萝芯具有较高的活性值(分别为914.34±25.47 U/mg和2152.36±75.99 U/mg)。这些结果与一维电泳和MALDI TOF - TOF蛋白质鉴定方法相结合,表明在新鲜木瓜和菠萝残渣提取物中先前报道的木瓜蛋白酶、菠萝蛋白酶和其他内肽酶的特征性信号肽存在。这些发现表明,通过对流干燥残渣不会改变蛋白水解酶的活性和结构。最后,证实使用20%硫酸铵作为沉淀剂,在不同工作规模下能够以更灵活的方式达到74%的效率,采用纯化和鉴定方案,使其成为哥伦比亚最有前景的废弃物,因为其具有大规模生产菠萝蛋白酶的潜力。

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