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Expression of a wheat β-1,3-glucanase in Pichia pastoris and its inhibitory effect on fungi commonly associated with wheat kernel.
- Source :
-
Protein Expression & Purification . Feb2019, Vol. 154, p134-139. 6p. - Publication Year :
- 2019
-
Abstract
- Abstract β -1,3-glucanases, the plant PR-2 family of pathogenesis-related (PR) proteins, can be constitutively expressed and induced in wheat crop to enhance its anti-fungal pathogen defense. This study aimed to investigate the inhibitory effect of wheat β -1,3-glucanase on fungi most commonly associated with wheat kernel. A β -1,3-glucanase from wheat was successfully expressed in Pichia pastoris X-33 and its biochemical and antifungal properties were characterized herein. The molecular weight of recombinant β -1,3-glucanase is approximately 33 kDa. β -1,3-glucanase displays optimal activity at pH 6.5, remaining relatively high at pH 5.5–8.0. The optimal reaction temperature of β -1,3-glucanase is 50 °C, retaining approximately 84.0% residual activity after heat-treated at 50 °C for 1 h. The steady-state kinetic parameters of β -1,3-glucanase against laminarin was determined and the K m and V max were 1.32 ± 0.20 mg/ml and 96.4 ± 4.4 U mg−1 protein, respectively. The inhibitory effect of purified β -1,3-glucanase against the seven fungi commonly associated with wheat kernel was assessed in vitro. β -1,3-glucanase exerted differential inhibitory effects on hyphal growth of Fusarium graminearum , Alternaria sp. , A. glaucus , A. flavus , A. niger , and Penicillium sp. Spore formation and mycelial morphology of Alternaria sp. , A. flavus , and A. niger were significantly affected by β -1,3-glucanase (1U). The present results would help elucidate the mechanism underlying the inhibition of wheat β -1,3-glucanases on pathogens. Highlights • A wheat β-1,3-glucanase was successfully expressed in Pichia pastoris X-33. • The biochemical and antifungal properties of wheat β-1,3-glucanase were characterized. • The β -1,3-glucanase has different inhibition effect on the seven fungi commonly associated with wheat kernel. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 154
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 133044247
- Full Text :
- https://doi.org/10.1016/j.pep.2018.10.011