Purification and characterization of an exo-polygalacturonase from Pycnoporus sanguineus

Mycological research Pub Date : 2009-12-01 DOI:10.1016/j.mycres.2009.09.007

Emma N. Quiroga , Melina A. Sgariglia , César F. Molina , Diego A. Sampietro , José R. Soberón , Marta A. Vattuone

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引用次数: 12

Abstract

The present work describes the purification and characterization of a novel extracellular polygalacturonase, PGase I, produced by Pycnoporus sanguineus when grown on citrus fruit pectin. This substrate gave enhanced enzyme production as compared to sucrose and lactose. PGase I is an exocellular enzyme releasing galacturonic acid as its principal hydrolysis product as determined by TLC and orcinol-sulphuric acid staining. Its capacity to hydrolyze digalacturonate identified PGase I as an exo-polygalacturonase. SDS-PAGE showed that PGase I is an N-glycosidated monomer. The enzyme has a molecular mass of 42 kDa, optimum pH 4.8 and stability between pH 3.8 and 8.0. A temperature optimum was observed at 50–60 °C, with some enzyme activity retained up to 80 °C. Its activation energy was 5.352 cal mol⁻¹. PGase I showed a higher affinity towards PGA than citric pectin (Km = 0.55 ± 0.02 and 0.72 ± 0.02 mg ml⁻¹, respectively). Consequently, PGase I is an exo-PGase, EC 3.2.1.82.

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血螺外聚半乳糖醛酸酶的纯化及特性研究

本工作描述了一种新的细胞外多半乳糖醛酸酶，PGase I的纯化和特性，该酶是由生长在柑橘果实果胶上的血蝎螺(Pycnoporus sanguineus)产生的。与蔗糖和乳糖相比，这种底物能提高酶的产量。通过薄层色谱法和orcinol-硫酸染色测定，PGase I是一种细胞外酶，其主要水解产物为半乳糖醛酸。它水解二半乳糖酸的能力鉴定了PGase I是一种外聚半乳糖酸酶。SDS-PAGE显示PGase I是一个n -糖苷化单体。酶的分子量为42 kDa，最适pH为4.8，稳定性在pH 3.8 ~ 8.0之间。在50-60°C的温度下，酶的活性保持在80°C。其活化能为5.352 cal mol−1。PGase I对PGA的亲和力高于柠檬酸果胶(Km分别为0.55±0.02和0.72±0.02 mg ml−1)。因此，PGase I是一个外显子PGase, EC 3.2.1.82。

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Mycological research

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