title: β-1,3-1,4-glucanase gene from Bacillus velezensis ZJ20 exerts antifungal effect on plant pathogenic fungi.
abstract: Bacillus velezensis is a known antifungal bacteria. To understand the role of β-1, 3-1, 4-glucanase played on B. velezensis about the mechanism which exerts effect on fungi, we isolated and cloned the β-1, 3-1, 4-glucanase gene (Bglu1) from B. velezensis ZJ20. The Bglu1 open reading frame was 732 bp that encoded a protein with 243 amino acids and a calculated molecular weight of 27.3 kDa. The same gene without the signal peptide, termed Bglu2, was also cloned and expressed in E. coli BL21. Among the two variants, only Bglu2 protein was expressed. Purified Bglu2 could be eluted with imidazole solution at concentrations ranging from 100 to 500 mM although the highest expression was observed at 150 and 200 mM and the purest was at 500 mM. In addition, activity of the crude enzyme was 1527 U ml(-1) and the highest activity of the purified enzyme was 1706 U ml(-1). The purified β-1, 3-1, 4-glucanase had activity on a wide range of pH and temperatures and displayed optimal activity at pH 5.0 and 35 °C. More importantly, the mycelial morphology of three pathogenic fungi was destroyed by the purified β-1, 3-1, 4-glucanase. In conclusion, β-1, 3-1, 4-glucanase from B. velezensis ZJ20 can be highly expressed in E. coli BL21 and the recombinant protein is pathogenic to fungi.
authors.affiliation: College of Forestry, Sichuan Agricultural University, Ya'an, 625014, China. sweetinxt@163.com.
authors.forename: Ting
authors.initials: T
authors.lastname: Xu
authors.affiliation: College of Forestry, Sichuan Agricultural University, Ya'an, 625014, China. zhuth1227@163.com.
authors.forename: Tianhui
authors.initials: T
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Forest Protection of Sichuan Province, Ya'an, 625014, China. lishujiangsumer@163.com.
authors.forename: Shujiang
authors.initials: S
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology*; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D001483:Base Sequence; D003001:Cloning, Molecular; D044644:Endo-1,3(4)-beta-Glucanase / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005658:Fungi / Q000166:cytology / Q000187:drug effects* / Q000472:pathogenicity; D015870:Gene Expression; D016366:Open Reading Frames; D010572:Pest Control, Biological / Q000379:methods; D010935:Plant Diseases / Q000382:microbiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D000935:Antifungal Agents; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D044644:Endo-1,3(4)-beta-Glucanase
keywords: Antifungal effect; Bacillus velezensis; Gene cloning; Protein expression; β-1,3-1,4-glucanase