Cloning, expression and characterization of a pectate lyase from Paenibacillus sp. 0602 in recombinant Escherichia coliReportar como inadecuado




Cloning, expression and characterization of a pectate lyase from Paenibacillus sp. 0602 in recombinant Escherichia coli - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

BMC Biotechnology

, 14:18

Protein and enzyme technology

Abstract

BackgroundBiotechnological applications of microbial pectate lyases Pels in plant fiber processing are considered as environmentally friendly. As such, they become promising substitutes for conventional chemical degumming process. Since applications of Pels in various fields are widening, it is necessary to explore new pectolytic microorganisms and enzymes for efficient and effective usage. Here, we describe the cloning, expression, characterization and application of the recombinant Pel protein from a pectolytic bacterium of the genus Paenibacillus in Escherichia coli.

ResultsA Pel gene pelN was cloned using degenerate PCR and inverse PCR from the chromosomal DNA of Paenibacillus sp. 0602. The open reading frame of pelN encodes a 30 amino acid signal peptide and a 445 amino acid mature protein belonging to the polysaccharide lyase family 1. The maximum Pel activity produced by E. coli in shake flasks reached 2,467.4 U mL, and the purified recombinant enzyme exhibits a specific activity of 2,060 U mg on polygalacturonic acid PGA. The maximum activity was observed in a buffer with 5 mM Ca at pH 9.8 and 65°C. PelN displays a half-life of around 9 h and 42 h at 50°C and 45°C, respectively. The biochemical treatment achieved the maximal reduction of percentage weight 30.5% of the ramie bast fiber.

ConclusionsThis work represents the first study that describes the extracellular expression of a Pel gene from Paenibacillus species in E. coli. The high yield of the extracellular overexpression, relevant thermostability and efficient degumming using combined treatments indicate its strong potential for large-scale industrial production.

KeywordsPectate lyase High-level expression Characterization Degumming Electronic supplementary materialThe online version of this article doi:10.1186-1472-6750-14-18 contains supplementary material, which is available to authorized users.

Xiaoman Li, Huilin Wang contributed equally to this work.

Download fulltext PDF



Autor: Xiaoman Li - Huilin Wang - Cheng Zhou - Yanhe Ma - Jian Li - Jiangning Song

Fuente: https://link.springer.com/







Documentos relacionados