Purification, characterization, and heterologous expression of an antifungal protein from the endophytic Bacillus subtilis strain Em7 and its activity against Sclerotinia sclerotiorum

被引：8

作者：

Wang, N. N. ^{[1
,2
]}

Gao, X. N. ^{[1
,2
]}

Yan, X. ^{[1
,2
]}

Li, Z. P. ^{[1
,2
]}

Kang, Z. S. ^{[1
,2
]}

Huang, L. L. ^{[1
,2
]}

Han, Q. M. ^{[1
,2
]}

机构：

[1] Northwest A&F Univ, State Key Lab Crop Stress Biol Arid Areas, Yangling, Shaanxi, Peoples R China

[2] Northwest A&F Univ, Coll Plant Protect, Yangling, Shaanxi, Peoples R China

来源：

GENETICS AND MOLECULAR RESEARCH | 2015年 / 14卷 / 04期

关键词：

Antifungal activity; Heterologous expression; Bacillus subtilis; beta-1,3-1,4-glucanase; Sclerotinia sclerotiorum; BETA-1,3-1,4-GLUCANASE GENE; BIOLOGICAL-CONTROL; BACILLOMYCIN-D; ANTIBIOTICS; CLONING; MYCOBACILLIN; LIPOPEPTIDES; MYCOSUBTILIN; PEPTIDE; DISEASE;

D O I：

10.4238/2015.November.30.27

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

An antifungal protein exhibiting a high activity against Sclerotinia sclerotiorum in vivo was purified by ammonium sulfate precipitation, hydrophobic chromatography, and gel filtration chromatography from the culture filtrate of the endophytic Bacillus subtilis strain Em7. The protein was characterized as a beta-1,3-1,4-glucanase according to amino acid analysis, and showed excellent properties in thermal stability and acid resistance. At the same time, the antifungal protein was cloned and heterologously expressed in Escherichia coli BL21. The recombinant protein was purified and showed similar enzymatic properties to the native protein, exhibiting strong inhibitory activity against S. sclerotiorum. This shows that the beta-1,3-1,4-glucanase may play a very important role in B. subtilis Em7 biocontrol function. In addition, many physiochemical properties of the native and purified recombinant protein were compared, including the effect of pH, temperature, metal cations, substrate specificity, and kinetic parameters. All parameters were similar between the native and recombinant purified protein, indicating that the purified recombinant protein has potential for industrial applications.

引用

页码：15488 / 15504

页数：17

共 39 条

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