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A Novel Multifunctional α-Amylase from the Thermophilic Fungus Malbranchea cinnamomea: Biochemical Characterization and Three-Dimensional Structure

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Abstract

A novel α-amylase (McAmyA) from the thermophilic fungus, Malbranchea cinnamomea was purified, characterized and crystallized in the present study. McAmyA was purified to apparent homogeneity with a molecular mass of 60.3 kDa on SDS-PAGE. The enzyme exhibited maximal activity at pH 6.5 and was stable within pH 5.0–10.0. It was most active at 65 °C and was stable up to 50 °C. McAmyA was capable of hydrolyzing amylose, starch, amylopectin, pullulan, cyclodextrins and maltooligosaccharides. The full-length cDNA of an α-amylase gene (McAmyA) from the strain was cloned. McAmyA consisted of a 1,476-bp open reading frame encoding 492 amino acids. It displayed the highest amino acid sequence homology (less than 60 %) with the reported α-amylases. The crystal structure of McAmyA was solved at a resolution of 2.25 Å (PDB code 3VM7). The overall structure of McAmyA reveals three domains with ten α helices and 14 β strands, and the putative catalytic residues are positioned at domain A with somewhat different secondary structural circumstances compared with typical α-amylases.

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Abbreviations

CAPS:

(Cyclohexylamino)-1-propanesulphonic acid

CD:

Cyclodextrin

CHES:

2-(cyclohexylamino) ethanesulfonic acid

GH:

Glycoside hydrolase

McAmyA:

An α-amylase from M. cinnamomea S168

MES:

2-(N-morpholino)ethane sulfonic acid

ORF:

Open reading frame

PCR:

Polymerase chain reaction

RACE:

Rapid amplification of cDNA ends

TLC:

Thin-layer chromatography

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Acknowledgments

This work was supported by a research grant from the National High Technology Research and Development Program of China (863 Program, no. 2011AA100905).

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Authors and Affiliations

  1. Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, PO Box 294, No.17 Qinghua Donglu, Haidian District, Beijing, 100083, China

    Peng Han, Peng Zhou, Shaoqing Yang & Zhengqiang Jiang

  2. College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, Guangdong, 510640, China

    Songqing Hu

  3. Bioresource Utilization Laboratory, College of Engineering, China Agricultural University, Beijing, 100083, China

    Qiaojuan Yan

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  1. Peng Han
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Correspondence to Zhengqiang Jiang.

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Han, P., Zhou, P., Hu, S. et al. A Novel Multifunctional α-Amylase from the Thermophilic Fungus Malbranchea cinnamomea: Biochemical Characterization and Three-Dimensional Structure. Appl Biochem Biotechnol 170, 420–435 (2013). https://doi.org/10.1007/s12010-013-0198-y

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