Identification and Functional Exploration of the Expansin Gene from Thermophilic Talaromyces leycettanus JCM12802

doi:10.13560/j.cnki.biotech.bull.1985.2018-0234

Abstract

Abstract: This study aims to obtain novel expansin-like genes,enrich expansin-like genes resources,explore its function,deepen our understanding of expansin-like and its mechanism of action,and promote its industrial application. In this study,an expansin-like gene Tlexlx1,1 196 bp in length,was cloned from thermophilic Talaromyces leycettanus JCM12802 by RT-PCR. In comparison to most expansin derived from fungi,TlEXLX1 lacks a CBM domain at the N-terminus. Deduced TlEXLX1 has the highest sequence similarities of 81% with the hypothetical protein ASPZODRAFT_140583 from Penicilliopsis zonata and 56% with the expansin-like protein 1 from Penicillium digitatum Pd1. Recombinant plasmids harboring CBM-TlEXLX1 with the CBM domain of TlSWOl at the N-terminus and wild TlEXLX1were constructed,their gene products were expressed in Pichia pastoris,and the basic features of their products were analyzed. This gene contained 1 intron (83 bp) and encodes a polypepetide of 370 amino acids and a stop codon. Deduced TlEXLX1 consisted of a 22-residue signal peptide at the N-terminus,a GH45 domain and an expansin-like domain. The purified recombinant TlEXLX1 and fusion protein CBM-TlEXLX1 had significant glucanase activities of degrading lichenan (7.2 and 17.2 U/mg,and barley β-glucan (4.4 and 9.4 U/mg),but weak hydrolytic activity of microcrystalline cellulose. Using lichenan as the substrate,both enzymes showed similar temperature and pH optima (60°C and 6.0). Moreover,TlEXLX1 had capability of destroying the neat and smooth surface structure of microcrystalline cellulose,and exhibited synergistic effect with commercial cellulase. Conclusively,a novel expansin obtained has hydrolytic activity and has potential value in the lignocellulose degradation industry..

Key words: expansin-like, Talaromyces leycettanus JCM12802, gene cloning, heterologous expression, CBM-fusion protein

GU Yuan, XUN Zi-qi, ZHENG Fei, TU Tao, YAO Bin, LUO Hui-ying. Identification and Functional Exploration of the Expansin Gene from Thermophilic Talaromyces leycettanus JCM12802[J]. Biotechnology Bulletin, 2018, 34(10): 172-181.

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