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Heterologous expression and periplasmic secretion of an antifungal Bacillus amyloliquefaciens BLB 369 endo‐β‐1,3‐1,4‐glucanase in Escherichia coli
Author(s) -
ZalilaKolsi Imen,
Sellami Sameh,
Tounsi Slim,
Jamoussi Kaïs
Publication year - 2018
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12656
Subject(s) - bacillus amyloliquefaciens , biology , escherichia coli , glucanase , periplasmic space , signal peptide , microbiology and biotechnology , molecular mass , biochemistry , heterologous expression , peptide sequence , gene , recombinant dna , enzyme , fermentation
The endo‐β‐1,3‐1,4‐glucanases are glycoside hydrolases involved in the enzymatic depolymerization of 1,3‐1,4 β‐glucans and showed an antifungal activity against some fungi. Bacillus amyloliquefaciens BLB 369 has a high antagonistic activity against phytopathogenic fungi. Its glu369 full‐coding sequence of the endo‐β‐1,3‐1,4‐glucanase gene (732 bp) was sequenced, cloned and successfully expressed in Escherichia coli Top10. The encoded protein (243 amino acids) has a calculated molecular mass of 27.3 kD a. To simplify the purification procedure, the glu369 coding sequence was cloned into the vector pKJD 4. The produced OmpA‐His‐Glu369 harboured OmpA signal sequence for E. coli periplasmic localization and followed by a 6His residues for its purification. The purified His‐tagged proteins revealed two bands on SDS ‐ PAGE analysis with molecular masses of about 30.5 (His‐Glu369) and 32.5 kD a (OmpA‐His‐Glu369). They had the ability to inhibit the growth of phytopathogenic fungus Alternaria alternata . These favourable properties make the endo‐β‐1,3‐1,4‐glucanase a good candidate for biotechnological applications.