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Recombinant Penicillium oxalicum 16 β-Glucosidase 1 Displays Comprehensive Inhibitory Resistance to Several Lignocellulose Pretreatment Products, Ethanol, and Salt

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Abstract

β-Glucosidase (BGL) is a rate-limiting enzyme of lignocellulose hydrolysis for second-generation bioethanol production, but its inhibition by lignocellulose pretreatment products, ethanol, and salt is apparent. Here, the recombinant Penicillium oxalicum 16 BGL 1 (rPO16BGL1) from Pichia pastoris GS115 kept complete activity at 0.2–1.4 mg/mL furan derivatives and phenolic compounds, 50 mg/mL sodium chloride (potassium chloride), or 100 mg/mL ethanol at 40 °C. rPO16BGL1 retained above 50% residual activity at 30 mg/mL organic acid sodium, and 60% residual activity at 40 °C with 300 mg/mL ethanol. Sodium chloride and potassium chloride had a complicated effect on rPO16BGL1, which resulted in activation or inhibition. The inhibition kinetics of the enzyme reaction demonstrated that organic acids and organic acid sodium were non-competitive inhibitors and that ethanol was a competitive inhibitor at < 1.5 mg/mL salicin. Moreover, substrate inhibition of the enzyme was found at > 2 mg/mL salicin, and the Km/KI and Km/KSI average values revealed that the inhibitory strength was ranked as salicin-organic acids > organic acids > salicin-organic acid sodium salt > organic acid sodium salt > salicin > salicin-KCl > salicin-NaCl > salicin-ethanol > ethanol.

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Funding

This work was financially supported by the National Natural Science Foundation of China [grant numbers 21666010, 31360217], the National Institutes of Health Training Program in Bioinformatics [grant number T32 GM070449], and the Doctoral Starting up Foundation of Jiangxi Normal University [grant number 5451].

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Authors and Affiliations

  1. College of Life Science, Jiangxi Normal University, Nanchang, 330022, China

    Hanxin Li, Shi Yi, Qiuxia Huang & Xihua Zhao

  2. Center for Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA

    Eric W. Bell

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  1. Hanxin Li
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Li, H., Yi, S., Bell, E.W. et al. Recombinant Penicillium oxalicum 16 β-Glucosidase 1 Displays Comprehensive Inhibitory Resistance to Several Lignocellulose Pretreatment Products, Ethanol, and Salt. Appl Biochem Biotechnol 191, 772–784 (2020). https://doi.org/10.1007/s12010-019-03183-y

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  • DOI: https://doi.org/10.1007/s12010-019-03183-y

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