Expression and biochemical characterization of two novel feruloyl esterases derived from fecal samples of Rusa unicolor and Equus burchelli

Thulasi, A.; Vijayarani, K.; Kumar, Duvvuri Prasanna; Santosh, Sunil Singh; Palanivel, C.; Jose, Vazhakkala Lyju; Sampath, K. T.

doi:10.1016/j.gene.2012.03.027

Cited by 13 publications

(1 citation statement)

References 22 publications

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“…Alkaline proteases have wide applications in different industries, such as detergent, feed processing, protein processing, and peptide synthesis (Chandrasekharaiah et al 2012;Adrio and Demain 2014). Many alkaline protease genes have been cloned, and their encoded proteases have been expressed, purified, and characterized (Deng et al 2010;Deng et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Cloning, secretory expression, partial characterization, and structural modeling of an alkaline protease from Bacillus subtilis D-2

Ding

et al. 2019

BioRes

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To develop a large-scale production of the protease of Bacillus subtilis strain D-2, the full-length gene apr-D2 (1,149 bp) encoding the alkaline protease was cloned into plasmid pET-32a and expressed as a secretory protein in Escherichia coli. Sequence analysis of the deduced amino acid sequence revealed high homology with the catalytic domains of the subtilisin serine proteases. From SDS-PAGE analysis, the recombinant protein had a molecular mass of 60.4 kDa. The expressed protease was secreted into the culture medium in a functional active form. The purified recombinant protease showed a pH optimum of 10.5 and temperature optimum of 55 °C, and it was stable in the pH range from 5.0 to 13.0. The enzyme activity was slightly enhanced by Ca2+, Mg2+, Ba2+, and SBT1. However, it was highly inhibited by Ag+ and PMSF. A theoretical structural model of mature protein was constructed by comparative modeling, which showed a putative catalytic triad (Asp-32, His-64 and Ser-221) with high similarity to the template. The structural characteristics that confer enzymatic specificity of the protease were analyzed. Taken together, the data suggested that the secretory expression system with pET-32a in E. coli was successfully constructed. Additionally, enzymatic specificity analysis of the alkaline protease indicated that it was suitable for various processing industries.

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