Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate

Li, Han-Zhi; Liu, Ming-Yu; Wang, Yu-Yue; Luo, Xue-Mei; Feng, Jia-Xun; Zhao, Shuai

doi:10.3390/ijms252211986

Open AccessArticle

Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate

by

Han-Zhi Li

^†,

Ming-Yu Liu

^†,

Yu-Yue Wang

,

Xue-Mei Luo

,

Jia-Xun Feng

^* and

Shuai Zhao

^*

State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi Research Center for Microbial and Enzyme Engineering Technology, College of Life Science and Technology, Guangxi University, Nanning 530004, China

^*

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Int. J. Mol. Sci. 2024, 25(22), 11986; https://doi.org/10.3390/ijms252211986

Submission received: 3 October 2024 / Revised: 29 October 2024 / Accepted: 3 November 2024 / Published: 7 November 2024

(This article belongs to the Section Biochemistry)

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Abstract

Rapeseed meal is severely restricted in its utilization as unconventional animal feed due to anti-nutritive compounds, such as glucosinolate, that are degraded to toxic nitriles such as 3-butenenitrile and 4-pentenenitrile in animals. Few studies on nitrilases that can degrade glucosinolate-derived nitriles have been reported thus far. In the present study, a nitrilase gene GiNIT from Gibberella intermedia was over-expressed in Escherichia coli and the purified recombinant nitrilase rGiNIT showed specific activities of 134.48 U/mg and 122.16 U/mg when using 3-butenenitrile and 4-pentenenitrile as substrates at the optimal pH, 7.5, and temperature, 45 °C, which is the highest reported in the literature. The conversion of 3-butenenitrile and 4-pentenenitrile by rGiNIT reached 81.89% and 80.23% after hydrolysis for 15 min and 300 min, respectively. Site-directed mutagenesis and molecular docking analysis revealed that the catalytic ability of rGiNIT depended on the substrate binding pocket comprising 13 key amino acid residues. These results provide a potential enzyme resource for rapeseed meal detoxification and theoretical guidance for protein engineering.

Keywords: nitrilase; nitriles; glucosinolate; rapeseed meal; substrate binding pocket

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MDPI and ACS Style

Li, H.-Z.; Liu, M.-Y.; Wang, Y.-Y.; Luo, X.-M.; Feng, J.-X.; Zhao, S. Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate. Int. J. Mol. Sci. 2024, 25, 11986. https://doi.org/10.3390/ijms252211986

AMA Style

Li H-Z, Liu M-Y, Wang Y-Y, Luo X-M, Feng J-X, Zhao S. Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate. International Journal of Molecular Sciences. 2024; 25(22):11986. https://doi.org/10.3390/ijms252211986

Chicago/Turabian Style

Li, Han-Zhi, Ming-Yu Liu, Yu-Yue Wang, Xue-Mei Luo, Jia-Xun Feng, and Shuai Zhao. 2024. "Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate" International Journal of Molecular Sciences 25, no. 22: 11986. https://doi.org/10.3390/ijms252211986

APA Style

Li, H.-Z., Liu, M.-Y., Wang, Y.-Y., Luo, X.-M., Feng, J.-X., & Zhao, S. (2024). Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate. International Journal of Molecular Sciences, 25(22), 11986. https://doi.org/10.3390/ijms252211986

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Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate

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