Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120

BMC Biotechnology
Hui QianGuang-Yu Yang

Abstract

Lipoxygenase (LOX) from Anabaena sp. PCC 7120 (Ana-rLOX) offers important applications in the food industry, especially for improving aroma and dough rheological properties. However, industrial applications of LOXs have been limited by their poor thermostability. Herein, we report a bioinformatics-based consensus concept approach for the engineering of thermostable Ana-rLOX. A series of mutations (N130D, G260A, S437T, N130D/G260Q, N130D/S437Y) showed higher thermostability and activity than the wild-type enzyme. Thus, N130D/G260Q exhibited a 6.6-fold increase in half-life and 2.45 °C increase in unfolding temperature; N130D/S437Y showed a 10 °C increase in optimal temperature. The secondary structure did not change much that contributed to improved thermostability were investigated in detail using circular dichroism. Homology modeling suggested that enhanced thermostability and specific activity may result from favorable hydrophobic interactions. A series of mutations were achieved, showing higher thermostability and activity than the wild-type enzyme by semi-rational mutagenesis with limited structure information. Our findings provide important new insights into molecular modifications aimed at improving Ana-rLOX thermostabili...Continue Reading

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Citations

Sep 5, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jiivittha VenoNor Hafizah Ahmad Kamarudin

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Methods Mentioned

BETA
circular dichroism
differential scanning calorimetry

Software Mentioned

ESpript
DICHROWEB
Basic Local Alignment Search Tool ( BLAST
Phyre2
ClustalX2
rLOX

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