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Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont
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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565265/ https://www.ncbi.nlm.nih.gov/pubmed/23344035 http://dx.doi.org/10.3390/ijms14010308 |
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author | Chang, Yu-Tzu Hsieh, Chienyan Wu, Li-Ching Chang, Hebron C. Kao, Suey-Sheng Meng, Menghsiao Hsieh, Feng-Chia |
author_facet | Chang, Yu-Tzu Hsieh, Chienyan Wu, Li-Ching Chang, Hebron C. Kao, Suey-Sheng Meng, Menghsiao Hsieh, Feng-Chia |
author_sort | Chang, Yu-Tzu |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3565265 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-35652652013-03-13 Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont Chang, Yu-Tzu Hsieh, Chienyan Wu, Li-Ching Chang, Hebron C. Kao, Suey-Sheng Meng, Menghsiao Hsieh, Feng-Chia Int J Mol Sci Article 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. MDPI 2012-12-21 /pmc/articles/PMC3565265/ /pubmed/23344035 http://dx.doi.org/10.3390/ijms14010308 Text en © 2013 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/). |
spellingShingle | Article Chang, Yu-Tzu Hsieh, Chienyan Wu, Li-Ching Chang, Hebron C. Kao, Suey-Sheng Meng, Menghsiao Hsieh, Feng-Chia Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont |
title | Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont |
title_full | Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont |
title_fullStr | Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont |
title_full_unstemmed | Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont |
title_short | Purification and Properties of an Insecticidal Metalloprotease Produced by Photorhabdus luminescens Strain 0805-P5G, the Entomopathogenic Nematode Symbiont |
title_sort | purification and properties of an insecticidal metalloprotease produced by photorhabdus luminescens strain 0805-p5g, the entomopathogenic nematode symbiont |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565265/ https://www.ncbi.nlm.nih.gov/pubmed/23344035 http://dx.doi.org/10.3390/ijms14010308 |
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