Lee Seong Hyuk, Kim Min-Sik, Kang Sung Gyun, Lee Hyun Sook
1Korea Institute of Ocean Science and Technology, Busan, 49111 Republic of Korea.
2Biomass and Waste Energy Laboratory, Korea Institute of Energy Research, Daejeon, 34129 Republic of Korea.
Biotechnol Biofuels. 2019 Feb 8;12:24. doi: 10.1186/s13068-019-1365-3. eCollection 2019.
The production of biohydrogen (H) as a promising future fuel in anaerobic hyperthermophiles has attracted great attention because H formation is more thermodynamically feasible at elevated temperatures and fewer undesired side products are produced. However, these microbes require anoxic culture conditions for growth and H production, thereby necessitating costly and time-consuming physical or chemical methods to remove molecular oxygen (O). Therefore, the development of an O-tolerant strain would be useful for industrial applications.
In this study, we found that the overexpression of -encoding hydrogenase genes in NA1, an obligate anaerobic archaeon and robust H producer, enhanced O tolerance. When the recombinant FO strain was exposed to levels of O up to 20% in the headspace of a sealed bottle, it showed significant growth. Whole transcriptome analysis of the FO strain revealed that several genes involved in the stress response such as chaperonin β subunit, universal stress protein, peroxiredoxin, and alkyl hydroperoxide reductase subunit C, were significantly up-regulated. The O tolerance of the FO strain enabled it to grow on formate and produce H under oxic conditions, where prior O-removing steps were omitted, such as the addition of reducing agent NaS, autoclaving, and inert gas purging.
Via the overexpression of genes, the obligate anaerobic archaeon NA1 gained the ability to overcome the inhibitory effect of O. This O-tolerant property of the strain may provide another advantage to this hyperthermophilic archaeon as a platform for biofuel H production.
在厌氧嗜热微生物中生产生物氢(H₂)作为一种未来有前景的燃料已引起广泛关注,因为在高温下H₂的形成在热力学上更可行,并且产生的不需要的副产物较少。然而,这些微生物生长和产生H₂需要无氧培养条件,因此需要昂贵且耗时的物理或化学方法来去除分子氧(O₂)。因此,开发一种耐氧菌株将对工业应用有用。
在本研究中,我们发现,在专性厌氧古菌且高效H₂生产者嗜热栖热菌NA1中过表达编码氢化酶的基因可增强其耐氧性。当重组FO菌株暴露于密封瓶顶空中高达20%的O₂水平时,它显示出显著生长。对FO菌株的全转录组分析表明,几个参与应激反应的基因,如伴侣蛋白β亚基、通用应激蛋白、过氧化物还原酶和烷基过氧化氢还原酶亚基C,均显著上调。FO菌株的耐氧性使其能够在甲酸盐上生长并在有氧条件下产生H₂,在此之前省略了诸如添加还原剂Na₂S、高压灭菌和惰性气体吹扫等除氧步骤。
通过基因的过表达,专性厌氧古菌嗜热栖热菌NA1获得了克服O₂抑制作用的能力。该菌株的这种耐氧特性可能为这种嗜热古菌作为生物燃料H₂生产平台提供另一个优势。