Engineering a fungal peroxidase that degrades lignin at very acidic pH

Biotechnology for Biofuels

Elena Fernández-Fueyo¹  Francisco J Ruiz-Dueñas²  Angel T Martínez² 

[1] Department of Biotechnology, TU Delft, Julianalaan 136, 2628 BL Delft, Netherlands

[2] Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, E-28040 Madrid, Spain

关键词: White-rot fungal genomes;    Versatile peroxidase;    Manganese peroxidase;    Lignin model dimer;    Catalytic tryptophan;    Acidic pH stability;   

Others  :  1084782
DOI  :  10.1186/1754-6834-7-114

 received in 2014-04-28, accepted in 2014-07-15,  发布年份 2014

【 摘 要 】

Background

Ligninolytic peroxidases are divided into three families: manganese peroxidases (MnPs), lignin peroxidases (LiPs), and versatile peroxidases (VPs). The latter two are able to degrade intact lignins, as shown using nonphenolic lignin model compounds, with VP oxidizing the widest range of recalcitrant substrates. One of the main limiting issues for the use of these two enzymes in lignocellulose biorefineries (for delignification and production of cellulose-based products or modification of industrial lignins to added-value products) is their progressive inactivation under acidic pH conditions, where they exhibit the highest oxidative activities.

Results

In the screening of peroxidases from basidiomycete genomes, one MnP from Ceriporiopsis subvermispora was found to have a remarkable acidic stability. The crystal structure of this enzyme recently became available and, after comparison with Pleurotus ostreatus VP and Phanerochaete chrysosporium LiP structures, it was used as a robust scaffold to engineer a stable VP by introducing an exposed catalytic tryptophan, with different protein environments. The variants obtained largely maintain the acidic stability and strong Mn²⁺-oxidizing activity of the parent enzyme, and the ability to oxidize veratryl alcohol and Reactive Black 5 (two simple VP substrates) was introduced. The engineered peroxidases present more acidic optimal pH than the best VP from P. ostreatus, enabling higher catalytic efficiency oxidizing lignins, by lowering the reaction pH, as shown using a nonphenolic model dimer.

Conclusions

A peroxidase that degrades lignin at very acidic pH could be obtained by engineering an exposed catalytic site, able to oxidize the bulky and recalcitrant lignin polymers, in a different peroxidase type selected because of its high stability at acidic pH. The potential of this type of engineered peroxidases as industrial biocatalysts in lignocellulose biorefineries is strongly enhanced by the possibility to perform the delignification (or lignin modification) reactions under extremely acidic pH conditions (below pH 2), resulting in enhanced oxidative power of the enzymes.

【 授权许可】

   
2014 Fernández-Fueyo et al.; licensee BioMed Central Ltd.

【 图 表 】

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