Abstract

Aspergillus fumigatus HBFH5 is a thermophilic fungus which can efficiently degrade lignocellulose and which produces a variety of glycoside hydrolase. In the present study, a novel β-mannanase gene (AfMan5A) was expressed in Pichia pastoris and characterized. AfMan5A is composed of 373 amino acids residues, and has a calculated molecular weight of 40 kDa. It has been observed that the amino acid sequence of AfMan5A showed 74.4% homology with the ManBK from Aspergillus niger. In addition, the recombined AfMan5A exhibited optimal hydrolytic activity at 60 °C and pH 6.0. It had no activity loss after incubation for 1h at 60 °C, while 65% of the initial activity was observed after 1 h at 70 °C. Additionally, it maintained about 80% of its activity in the pH range from 3.0 to 9.0. When carob bean gum was used as the substrate, the Km and Vmax values of AfMan5A were 0.21 ± 0.05 mg·mL−1 and 15.22 ± 0.33 U mg−1·min−1, respectively. AfMan5A and AfSwol showed a strong synergistic interaction on galactomannan degradation, increasing the reduction of the sugars by up to 31%. Therefore, these findings contribute to new strategies for improving the hydrolysis of galactomannan using the enzyme cocktail.

Highlights

  • Mannans are one of the significant hemicelluloses, and are an inexpensive and sustainable source of renewable energy, functional food and other uses [1,2]

  • A. fumigatus, which is widely distributed in nature, has received attention as an excellent producer of glycoside hydrolase, including amylase, tannase and cellulase

  • Some excellent β-mannanase obtained from A. fumigatus has been reported

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Summary

Introduction

Mannans are one of the significant hemicelluloses, and are an inexpensive and sustainable source of renewable energy, functional food and other uses [1,2]. They are composed of mannose, glucose and galactose by linked β-1, 4 or α-1, 6 glycosidic bonds [3,4]. The most widely known β-mannanase belong to the GH5 and GH26 families, and some of them have been cloned and functionally characterized [8] These enzymes showed optimal temperatures around 40–60 ◦ C and optimal pH around 5.0–8.0, respectively [9]. Poor tolerance and low catalytic efficiency are serious constraints to the application of β-mannanase

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