Hydrogenase-catalyzed H₂/air biofuel cells have attracted wide attention. However, how to obtain thermostable and highly active hydrogenase remains challenging. Herein, we developed a thermophilic archaea host system to heterologously express a thermostable Pyrococcus furiosus cytoplasmic [NiFe]-hydrogenase I (PfSHI). The recombinant hydrogenase could be easily isolated and then immobilized onto a carbon nanotube-modified carbon felt electrode for further electrochemical characterization. The bioanode was able to work at a wide range of temperatures from 40 °C to 80 °C and no high potential deactivation was observed until 0.3 V at 50 °C. Cyclic voltammograms at elevated temperatures for reverse H₂ oxidation performance indicated that an inactive state might be formed and then reversed when scanning to negative position. Also, the power density of a whole cell reached 1.08 mW cm⁻² using Pt/C as the cathode with a brilliant open circuit potential reaching 1.2 V. The voltage could retain 90% of its initial value after 33 h discharge at a current of 10 μA. Besides, the bioanode exhibited decent oxygen tolerance both in ambient and elevated temperatures. These results suggest that this recombinant PfSHI can be a good candidate for catalyzing H₂/air biofuel cells.

氢酶催化的H ₂ /空气生物燃料电池引起了广泛的关注。然而，如何获得热稳定的和高活性的氢化酶仍然具有挑战性。在这里，我们开发了嗜热古细菌宿主系统，以异源表达热稳定的激烈热球菌胞质[NiFe]-氢化酶I（Pf SHI）。可以容易地分离重组氢化酶，然后将其固定在碳纳米管修饰的碳毡电极上，以进行进一步的电化学表征。生物阳极能够在40°C至80°C的宽温度范围内工作，直到在50°C达到0.3 V时才观察到高电位失活。高温下反向H _2的循环伏安图氧化性能表明，当扫描到负位置时，可能会形成惰性状态，然后反转。同样，使用Pt / C作为阴极，整个电池的功率密度达到1.08 mW cm ^-2，其开路电势达到1.2V。经过33 h的电流放电后，电压可以保持其初始值的90％。 10微安 此外，生物阳极在环境温度和高温下均表现出良好的耐氧性。这些结果表明，该重组Pf SHI可以是催化H ₂ /空气生物燃料电池的良好候选者。