Green-synthesized nickel oxide nanoparticles enhances biohydrogen production of Klebsiella sp. WL1316 using lignocellulosic hydrolysate and its regulatory mechanism

Abstract

Green synthesis of nickel oxide nanoparticle (NiO-NP) from Eichhornia crassipes (Ec) extract was novelly conducted, and regulatory effect of this NP on fermentative hydrogen production was evaluated. Characterization of the Ec-NiO-NP revealed their cubic and spherical shape, small diameter (9.1 ± 2.6 nm) and high purity. The maximum cumulative hydrogen production and hydrogen yield Y(H2/S) reached 4842.19 ± 23.43 mL/L and 101.45 ± 3.32 mL/gsubstrate, respectively, in the presence of 20 mg/L Ec-NiO-NP, which were 47.29% and 37.78% higher than the control without NPs addition. Evaluation of glucose and xylose utilization efficiency as well as key node metabolites further established the potential of Ec-NiO-NP to improve the reducing sugar conversion and metabolic flux distribution in hydrogen synthesis pathway. Furthermore, addition of 20 mg/L Ec-NiO-NP resulted in enhanced hydrogenase activity with a maximum increase of 623% comparing to the control, and led to changes in the gene expression of both hydrogenase and formate-hydrogen lyase, which play important roles in promoting hydrogen production at different stages of fermentation. This study demonstrates that supplementation with green-synthesized Ec-NiO-NP effectively improves fermentative hydrogen production and regulates key node metabolites alteration and functional gene expression.