Identification and biochemical analysis of a novel pectate lyase 3 gene in Bursaphelenchus xylophilus

• Published in: Journal of Asia-Pacific entomology Vol. 16; no. 3; pp. 335 - 342
• Main Authors: Lee, Dae-Weon, Kang, Jae Soon, Jung, Chan Sik, Han, Hye Rhym, Moon, Yil Sung, Park, Seok Ju, Lee, Si Hyeock, Koh, Young Ho
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.09.2013; 한국응용곤충학회
• Subjects: AGENT PATHOGENE; Bacillus; BURSAPHELENCHUS XYLOPHILUS; calcium; catalytic activity; cell walls; expressed sequence tags; Fluorescence in situ hybridization; ORGANISMOS PATOGENOS; Pathogen; pathogenesis; PATHOGENS; pectate lyase; Pectate lyase 3; petate lyase 3,pine wilt disease,Fluorescence in situ hybridization; Pine wilt disease; quantitative polymerase chain reaction; secretion; stylets; 농수해양학; Pine wilt disease; Pathogen; Pectate lyase 3; Bursaphelenchus xylophilus; Fluorescence in situ hybridization
• ISSN: 1226-8615; 1876-7990
• DOI: 10.1016/j.aspen.2013.04.016

A novel pectate lyase 3 (Bx-PEL3) gene from the stage-specifically expressed sequence tag library of Bursaphelenchus xylophilus was cloned. Bx-PEL1 and Bx-PEL2 were relatively predominant to Bx-PEL3 in quantitative real-time PCR analysis. The variation of expression level among Bx-PELs according to different life stages suggests that each Bx-PEL may play different biochemical roles in pathogenesis. Recombinant Bx-PEL3 showed activity against polygalacturonic acid and its optimized physical conditions for pH and Ca2+ concentrations were 9.0 and 0.5mM, respectively. Homology modeling revealed that Bx-PELs were structurally similar to PEL from the Bacillus strain KSM P-15 and shared essential residues for calcium binding and catalytic activity. Bx-PEL3 was expressed near the esophageal gland as shown in other Bx-PELs, indicating that Bx-PEL3 may be involved in the degradation of the cell wall after secretion from the stylet. Our data suggest that the novel PEL3 gene is a putative pathogenic factor which is biochemically functional. [Display omitted] •A novel pectate lyase 3 was cloned from EST library of pinewood nematode.•Pectate lyase 3 was expressed at the stylet of the nematode.•Bacteria expressed pectate lyase 3 was functional.•3-D structure of pectate lyase 3 was similar to pectate lyase from Bacillus subsp. KSM P-15.