Identification and Characterization of an Anti-fungi Fusarium oxysporum f. sp. cucumerium Protease from the Bacillus subtilis Strain N7

Yi Luo; Lifei Sun; Zhen Zhu; Wei Ran; Qirong Shen

doi:10.1007/s12275-013-2627-6

Articles

Page Path: HOME > J. Microbiol > Volume 51(3); 2013 > Article

Research Support, Non-U.S. Gov't
Identification and Characterization of an Anti-fungi Fusarium oxysporum f. sp. cucumerium Protease from the Bacillus subtilis Strain N7: Yi Luo , Lifei Sun , Zhen Zhu , Wei Ran , Qirong Shen; Journal of Microbiology 2013;51(3):359-366.
DOI: https://doi.org/10.1007/s12275-013-2627-6
Published online: June 28, 2013

Jiangsu Key Lab for Organic Solid Waste Utilization, Nanjing Agricultural University, Nanjing 210095, P. R. ChinaJiangsu Key Lab for Organic Solid Waste Utilization, Nanjing Agricultural University, Nanjing 210095, P. R. China

Corresponding author: Wei Ran , Tel: +86-025-84396212,

Received: 14 November 2012 • Accepted: 4 February 2013

prev next

57 Views
0 Download
11 Crossref
14 Scopus

Full Article

Abstract

A newly discovered alkaline antifungal protease named P6 from Bacillus subtilis N7 was purified and partially characterized. B. subtilis N7 culture filtrates were purified by 30–60% (NH4)2SO4 precipitation, anion-exchange chromatography and gel filtration chromatography. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) revealed a single band of 41.38 kDa. Peptide sequence of protease P6 was determined using a 4800 Plus MALDI TOF/TOFTM Analyzer System. Self-Formed Adaptor PCR (SEFA-PCR) was used to amplify the 1,149 bp open read frame of P6. Dimensional structure prediction using Automatic Modeling Mode software showed that the protease P6 consisted of two β-barrel domains. Purified P6 strongly inhibited spore and mycelium growth of Fusarium oxysporum f. sp. cucumerium (FOC) by causing hypha lysis when the concentration was 25 μg/ml. Characterization of the purified protease indicated that it had substrate specificity for gelatin and was highly active at pH 8.0–10.6 and 70°C. The P6 protease was inhibited by EDTA (2 mmol/L), phenyl methyl sulfonyl fluoride (PMSF, 1 mmol/L), Na+, Fe3+, Cu2+, Mg2+ (5 mmol/L each) and H2O2 (2%, v/v). However, protease activity was activated by Ca2+, K+, Mn2+ (5 mmol/L each), mercaptoethanol (2%, v/v) and Tween 80 (1%, v/v). In additon, activity was also affected by organic solvents such as acetone, normal butanol and ethanol, but not hexane (25%, v/v each).

Keywords: purification;characterization;Bacillus subtilis;anti-fungal protease;self-formed adaptor PCR

Figure & Data

References

Citations

Citations to this article as recorded by

Mechanisms of Surfactin from Bacillus subtilis SF1 against Fusarium foetens: A Novel Pathogen Inducing Potato Wilt
Lin Liu, Xiaofan Jin, Xiuhua Lu, Lizhong Guo, Peiwei Lu, Hao Yu, Beibei Lv
Journal of Fungi.2023; 9(3): 367. CrossRef
The Extracellular Lipopeptides and Volatile Organic Compounds of Bacillus subtilis DHA41 Display Broad-Spectrum Antifungal Activity against Soil-Borne Phytopathogenic Fungi
Dhabyan Mutar Kareem Al-Mutar, Muhammad Noman, Noor Salih Abduljaleel Alzawar, Hadi Hussein Qasim, Dayong Li, Fengming Song
Journal of Fungi.2023; 9(8): 797. CrossRef
An Overview of Proteases: Production, Downstream Processes and Industrial Applications
Nathiele Contrera Gimenes, Edgar Silveira, Elias Basile Tambourgi
Separation & Purification Reviews.2021; 50(3): 223. CrossRef
Biocontrol of Orchid-pathogenic Mold,Phytophthora palmivora, by Antifungal Proteins fromPseudomonas aeruginosaRS1
Rapeewan Sowanpreecha, Panan Rerngsamran
Mycobiology.2018; 46(2): 129. CrossRef
Microbial Flora Associated with the Halophyte–Salsola imbricate and Its Biotechnical Potential
Fehmida Bibi, Gary A. Strobel, Muhammad I. Naseer, Muhammad Yasir, Ahmed A. Khalaf Al-Ghamdi, Esam I. Azhar
Frontiers in Microbiology.2018;[Epub] CrossRef
Diversity of indigenous endophytic bacteria associated with the roots of Chinese cabbage (Brassica campestris L.) cultivars and their antagonism towards pathogens
Md. Azizul Haque, Han Dae Yun, Kye Man Cho
Journal of Microbiology.2016; 54(5): 353. CrossRef
Isolation and anti- Verticillium dahliae activity from Bacillus axarquiensis TUBP1 protein
Hong Zeng, Rong Chen, Xiaoxia Luo, Jun Tian
Process Biochemistry.2016; 51(10): 1691. CrossRef
Effect of Two Biological Formulations Based onBacillus subtilisandPseudomonas fluorescenson Control ofDidymella applanata, the Causal Agent of Red Raspberry Cane Spur Blight
Margarita Shternshis, Tatyana Shpatova, Anatoly Belyaev
International Journal of Agronomy.2016; 2016: 1. CrossRef
Cucumber Rhizosphere Microbial Community Response to Biocontrol Agent Bacillus subtilis B068150
Lihua Li, Jincai Ma, A. Ibekwe, Qi Wang, Ching-Hong Yang
Agriculture.2015; 6(1): 2. CrossRef
Construction of a heterologous gene expression system in the banana rhizobacterium strain GW-3 and its colonization ability
Yuguang Wang, Qiyu Xia, He Zhang, Xuehua Lu, Jianbo Sun, Xin Zhang
World Journal of Microbiology and Biotechnology.2014; 30(3): 903. CrossRef
A novel antifungal protein of Bacillus subtilis B25
Zhiqiong Tan, Baoying Lin, Rongyi Zhang
SpringerPlus.2013;[Epub] CrossRef

Cite this Article

Cite this Article: export Copy Download Format; Close

Download Citation

Download a citation file in RIS format that can be imported by all major citation management software, including EndNote, ProCite, RefWorks, and Reference Manager.

Format:

RIS — For EndNote, ProCite, RefWorks, and most other reference management software
BibTeX — For JabRef, BibDesk, and other BibTeX-specific software

Include:

Citation for the content below
Citation and abstract for the content below

Identification and Characterization of an Anti-fungi Fusarium oxysporum f. sp. cucumerium Protease from the Bacillus subtilis Strain N7

J. Microbiol. 2013;51(3):359-366. Published online June 28, 2013

DOI: https://doi.org/10.1007/s12275-013-2627-6

Related articles