Int. J. Mol. Sci. 2013, 14(1), 308-321; doi:10.3390/ijms14010308
Article

Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont

Yu-Tzu Chang 1 email, Chienyan Hsieh 2 email, Li-Ching Wu 3 email, Hebron C. Chang 1 email, Suey-Sheng Kao 3 email, Menghsiao Meng 4,* email and Feng-Chia Hsieh 3,* email
1 Institute of Biotechnology and Bioinformatics, Asia University, Wufeng, Taichung 413, Taiwan 2 Department of Biotechnology, National Kaohsiung Normal University, Kaohsiung 824, Taiwan 3 Biopesticides Division, Taiwan Agricultural Chemicals and Toxic Substances Research Institute, Council of Agriculture, Wufeng, Taichung 413, Taiwan 4 Graduate Institute of Biotechnology, National Chung Hsing University, Taichung 402, Taiwan
* Authors to whom correspondence should be addressed.
Received: 29 October 2012; in revised form: 3 December 2012 / Accepted: 4 December 2012 / Published: 21 December 2012
(This article belongs to the Special Issue Green Biocides)
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Abstract: A total of 13 Photorhabdus luminescens strains were screened for proteolytic activity. The P. luminescens strain 0805-P5G had the highest activity on both skim milk and gelatin plates. The protease was purified to electrophoretical homogeneity by using a two-step column chromatographic procedure. It had a molecular weight of 51.8 kDa, as determined by MALDI-TOF mass spectrometry. The optimum pH, temperature, as well as pH and thermal stabilities were 8, 60 °C, 5–10, and 14–60 °C, respectively. It was completely inhibited by EDTA and 1,10-phenanthroline. Bioassay of the purified protease against Galleria mellonella by injection showed high insecticidal activity. The protease also showed high oral toxicity to the diamondback moth (Plutella xylostella) of a Taiwan field-collected strain, but low toxicity to an American strain. To our knowledge, this is the first report to demonstrate that the purified protease of P. luminescens has direct toxicity to P. xylostella and biopesticide potentiality.
Keywords: protease; Photorhabdus luminescens; Plutella xylostella; insecticidal activity

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MDPI and ACS Style

Chang, Y.-T.; Hsieh, C.; Wu, L.-C.; Chang, H.C.; Kao, S.-S.; Meng, M.; Hsieh, F.-C. Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont. Int. J. Mol. Sci. 2013, 14, 308-321.

AMA Style

Chang Y-T, Hsieh C, Wu L-C, Chang HC, Kao S-S, Meng M, Hsieh F-C. Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont. International Journal of Molecular Sciences. 2013; 14(1):308-321.

Chicago/Turabian Style

Chang, Yu-Tzu; Hsieh, Chienyan; Wu, Li-Ching; Chang, Hebron C.; Kao, Suey-Sheng; Meng, Menghsiao; Hsieh, Feng-Chia. 2013. "Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont." Int. J. Mol. Sci. 14, no. 1: 308-321.

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