Extracellular novel metalloprotease from Xenorhabdus indica and its potential as an insecticidal agent.
Proteases produced by Xenorhabdus are known to play a significant role in virulence leading to insect mortality. The present study was undertaken to purify and characterize protease from Xenorhabdus indica, an endosymbiont of nematode Steinernema thermophilum, and to decipher its role in insect mortality and its efficacy to control Helicoverpa armigera. A set of 10 strains of Xenorhabdus isolated from different regions of India were screened for protease activity on the basis of zone of clearing on gelatin agar plates. One potent strain of Xenorhabdus indica was selected for the production of protease, and the highest production (1,552 U/ml) was observed at 15-18 h of incubation at 28°C in soya casein digest broth. The extracellular protease was purified from culture supernatant using ammonium sulfate precipitation and ion-exchange chromatography. The enzyme was further characterized by SDS-PAGE and zymography, which confirmed the purity of the protein and its molecular mass was found to be ~52 kDa. Further MALDI-TOF/TOF analysis and effect of metal chelating agent 1,10-phenanthrolin study revealed the nature of the purified protease as a secreted alkaline metalloprotease. The bioefficacy of the purified protease was also tested against cotton bollworm (Helicoverpa armigera) and resulted in 67.9 ± 0.64% mortality within one week. This purified protease has the potential to be developed as a natural insecticidal agent against a broad range of agriculturally important insects.
Division of Microbiology, Indian Agricultural Research Institute, New Delhi-110012, India.
Kumar
K
Pranaw
Surender
S
Singh
Debjani
D
Dutta
Nirpendra
N
Singh
Garima
G
Sharma
Sudershan
S
Ganguly
Vinay
V
Kalia
Lata
L
Nain
D000818:Animals; D001681:Biological Assay; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D007194:India; D007306:Insecticides / Q000302:isolation & purification* / Q000378:metabolism*; D007915:Lepidoptera / Q000187:drug effects* / Q000502:physiology; D045726:Metalloproteases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008970:Molecular Weight; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D016019:Survival Analysis; D020593:Xenorhabdus / Q000201:enzymology* / Q000254:growth & development / Q000302:isolation & purification
D003470:Culture Media; D007306:Insecticides; D045726:Metalloproteases