Cloning and overexpression of a new chitosanase gene from Penicillium sp. D-1.

AMB Express
Xu-Fen ZhuMin Wu

Abstract

A chitosanase gene, csn, was cloned from Penicillium sp. D-1 by inverse PCR. The cDNA sequence analysis revealed that csn had no intron. The deduced CSN protein consists of 250 amino acids including a 20-amino acid signal peptide, and shared 83.6% identity with the family 75 chitosanase from Talaromyces stipitatus (B8M2R4). The mature protein was overexpressed in Escherichia coli and purified with the affinity chromatography of Ni2+-NTA. The novel recombinant chitosanase showed maximal catalytic activity at pH 4.0 and 48°C. Moreover, the activity of CSN was stable over a broad pH range of 3.0-8.0, and the enzymatic activity was significantly enhanced by Mg2+ and Mn2+. The CSN could effectively hydrolyze colloidal chitosan and chitosan, while could not hydrolyze chitin and carboxymethylcellulose (CMC). Due to the particular acidophily, CSN has the potential application in the recycling of chitosan wastes.The GenBank/EMBL/DDBJ accession numbers for the 18S rRNA gene and chitosanase gene of strain D-1 are JF950269 and JF950270, respectively.

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Citations

Oct 4, 2015·International Journal of Biological Macromolecules·Nathália Kelly de AraújoEveraldo Silvino Dos Santos
Jul 7, 2015·Biochimica Et Biophysica Acta·Qianqian LyuWeizhi Liu
Dec 24, 2013·Food Chemistry·Nidheesh Thadathil, Suresh Puthanveetil Velappan
Jan 19, 2019·World Journal of Microbiology & Biotechnology·Gleb E AktuganovVioletta R Safina
Dec 19, 2020·Comprehensive Reviews in Food Science and Food Safety·Huihui SunXiangzhao Mao
Aug 19, 2015·International Journal of Biological Macromolecules·Jianping ZhangHeng Yin
Apr 4, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yuanxiang PangYuanhong Xie
Jun 26, 2021·International Journal of Biological Macromolecules·Beata Kaczmarek-SzczepańskaMaria Swiontek Brzezinska

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Datasets Mentioned

BETA
JF950269
JF950270

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

BLAST
Mega
Primer Premier
Clustal X
SignalP
BoxShade

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