Cloning and Bioinformatics Analysis of Cellobiase Gene from Aspergillus niger C112

doi:10.13560/j.cnki.biotech.bull.1985.2016.02.015

Abstract

Abstract: The cellobiase gene(GenBank accession number：KP307454)was cloned by RT-PCR from high-yield cellulase Aspergillus nigerstrain C112 mutated by UV.The whole length of cellobiase was 2 934 bp containing non-coding sequence, and encoding 860 amino acids with the pI of 4.70.The homology of nucleotide sequence with Aspergillus niger(GenBank accession number：JX982101.1)reached 99%.Bioinformatics analysis showed that cellobiase was a hydrophilic, stable, and secreted protein；its secondary structure consisted of the structural elements of α-helix, β-folding and random coil.As expected, recombinant fusion protein was expressed after IPTG induction, and the relative molecular mass was approximately 93.3 kD by SDS-PAGE.The fused β-Glucosidase was expressed in Escherichia coli BL21.The enzyme activity of recombinant protein pNPGwas 5.847 U/mL, the optimal reaction temperature was 50℃, and the optimal reaction pH was 5.0.

Key words: Aspergillus niger C112, cellobiase, cloning, bioinformatics

ZHU Yong-rui, ZENG Bai-quan,ZENG Lei, LIU Hui. Cloning and Bioinformatics Analysis of Cellobiase Gene from Aspergillus niger C112[J]. Biotechnology Bulletin, 2016, 32(2): 116-122.

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