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. 2021 May 28;11(1):77.
doi: 10.1186/s13568-021-01236-2.

Effects of cofD gene knock-out on the methanogenesis of Methanobrevibacter ruminantium

Jian Ma #  1 , Xueying Wang #  1 , Ting Zhou  1 , Rui Hu  1 , Huawei Zou  1 , Zhisheng Wang  2 , Cui Tan  1 , Xiangfei Zhang  1 , Quanhui Peng  1 , Bai Xue  1 , Lizhi Wang  1
Affiliations

Effects of cofD gene knock-out on the methanogenesis of Methanobrevibacter ruminantium

Jian Ma et al. AMB Express. .

Abstract

This study aimed to investigate the effects of cofD gene knock-out on the synthesis of coenzyme F420 and production of methane in Methanobrevibacter ruminantium (M. ruminantium). The experiment successfully constructed a cofD gene knock-out M. ruminantium via homologous recombination technology. The results showed that the logarithmic phase of mutant M. ruminantium (12 h) was lower than the wild-type (24 h). The maximum biomass and specific growth rate of mutant M. ruminantium were significantly lower (P < 0.05) than those of wild-type, and the maximum biomass of mutant M. ruminantium was approximately half of the wild-type; meanwhile, the proliferation was reduced. The synthesis amount of coenzyme F420 of M. ruminantium was significantly decreased (P < 0.05) after the cofD gene knock-out. Moreover, the maximum amount of H2 consumed and CH4 produced by mutant were 14 and 2% of wild-type M. ruminantium respectively. In conclusion, cofD gene knock-out induced the decreased growth rate and reproductive ability of M. ruminantium. Subsequently, the synthesis of coenzyme F420 was decreased. Ultimately, the production capacity of CH4 in M. ruminantium was reduced. Our research provides evidence that cofD gene plays an indispensable role in the regulation of coenzyme F420 synthesis and CH4 production in M. ruminantium.

Keywords: Coenzyme F420; Gene knock-out; M ethanobrevibacter ruminantium; Methane; cofD gene.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
The differences of mRNA expression (mean and SD) of cofD in WT and KO M. ruminantium. Strains were incubated at 30° C for 72 h. SD standard deviation, WT wild-type, KO knock-out. The asterisk (*) indicates a significant difference (P < 0.05)
Fig. 2
Fig. 2
The differences protein expression (mean) of cofD in WT and KO M. ruminantium. Strains were incubated at 30° C for 72 h. WT wild-type, KO knock-out. The asterisk (*) indicates a significant difference (P < 0.05)
Fig. 3
Fig. 3
Growth kinetics of WT and cofD KO strains of M. ruminantium grown in BJ medium at 39 ℃ (n = 3). Growth was determined by measuring the absorbance of the cultures at 600 nm. WT wild-type, KO knock-out
Fig. 4
Fig. 4
The content (mean and SD) of coenzyme F420 of WT and KO M. ruminantium in different culture time. A the content of coenzyme F420 of liquid; B the content of coenzyme F420 per OD600. SD standard deviation, WT wild-type, KO knock-out
Fig. 5
Fig. 5
The concentration changes of methane production and hydrogen consumption in WT and KO cofD type M. ruminantium at 5 time points (12, 24, 36, 48, 96 h) (n = 3). WT wild-type, KO knock-out
See this image and copyright information in PMC

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