Identification and characterization of a novel endo-β-1,4-glucanase from a soil metagenomic library

• Published in: Carbohydrate research 2021-12, Vol.510, p.108460-108460, Article 108460
• Main Authors: Chai, Shumao, Zhang, Xueliang, Gao, Yuejiao, Nie, Lishuang, Xiang, Chenchen, Cao, Mingming, Wang, Shaochen, Feng, Zhiyang
• Format: Article
• Language: English
• Subjects: endo-β-1,4-Glucanase; Functional metagenomics; Gene Library; Glycoside hydrolase family 6; Glycoside Hydrolases - genetics; Glycoside Hydrolases - metabolism; Metagenomics; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Soil; Transmembrane region
• ISSN: 0008-6215; 1873-426X
• DOI: 10.1016/j.carres.2021.108460

A cosmid clone cZFYN1413 with CMCase activity was identified from a soil metagenomic library. The sequence analysis of a subclone of cZFYN1413 revealed an endo-β-1,4-glucanase gene ZFYN1413 belonging to glycoside hydrolase family 6 and a transmembrane region in the N-terminal of ZFYN1413. Expression of ZFYN1413 in Escherichia coli BL21 (DE3) resulted in ZFYN1413-87, which was a truncated protein cleaved in transmembrane region of ZFYN1413. ZFYN1413-87 was expressed and its enzyme properties were studied. ZFYN1413-87 possessed strong endo-β-1,4-glucanase activity, and 52% of the activity could be retained after the protein was treated in buffer of pH 3.0 for 2 h. The study provided a special example of endo-β-1,4-glucanase in GH6 family. [Display omitted] •A novel endo-β-1,4-glucanase was identified from a soil metagenomic library.•The enzyme is stable at pH 3.0-9.0 and 20-40°C.•Cellulase self-cleaved from the transmembrane region during the protein processing.•The novel enzyme provides a special example of cellulase in glycoside hydrolase family 6.