Effect of Carbon Source in Methanogenic Communities Regulation and Methane Production Capacity

doi:10.13560/j.cnki.biotech.bull.1985.2024-1180

Abstract

Abstract:

Objective Methanogens play a pivotal role in methanogenesis during anaerobic digestion. To obtain an efficient methanogenic microbial community, the regulatory effects of different carbon sources on the methane production potential, methanogen abundance, and microbial community structure were investigated. Method Three distinct carbon source culture media (Group A: CH₃COONa and CH₃OH;Group B: CH₃COONa; Group C: CH₃COONa and HCOONa) were designed to enrich methanogenic communities by inoculating with anaerobic digestion liquid from kitchen waste (FW) and activated sludge suspension (SS). The performance and diversity composition of methanogenic microbial community were analyzed using gas chromatography, real-time quantitative PCR (qPCR), ﬂuorescence microscopy, and amplicon sequencing. Result The microbial communities enriched by carbon sources A, B, and C demonstrated a descending order of methane yield when inoculated with the same inoculum. Group 1A-2 achieved the highest cumulative methane yield over 49 d, with a methane yield of 35.9 mL per gram of available carbon (C) in a unit volume (L) of culture medium. The copy number of mcrA gene and fluorescence microscopy observations indicated a successive decrease in methanogen abundance in communities enriched by carbon sources A, B, and C. Specifically, carbon source A enriched methylotrophic methanogens, while carbon source B significantly boosted the relative abundance of aceticlastic Methanothrix compared to the other groups. Carbon source C notably enhanced the relative abundance of hydrogenotrophic methanogens. Conclusion The type of carbon source significantly influence the diversity, richness, and community structure of methanogens and bacteria in the enriched microbial communities. The microbial communities enriched with CH₃COONa and CH₃OH (Group A) showed the highest methanogen abundance, mainly including Methanothrix, Methanoculleus, Methanomethylovorans, Candidatus Methanoplasmaand Methanosarcina, and demonstrated the highest methanogenic capacity. Carbon source A promotes methane (CH₄) production by enriching syntrophic bacteria, balancing the metabolic pathway distribution of methanogens, and facilitating direct interspecies electron transfer (DIET).

Key words: anaerobic culture, methanogens, real-time quantitative PCR, fluorescence observation, amplicon sequencing

AN Miao-miao, LIN Xiang-hao, LIANG Rui-na, ZHAO Guo-zhu. Effect of Carbon Source in Methanogenic Communities Regulation and Methane Production Capacity[J]. Biotechnology Bulletin, 2025, 41(6): 355-366.

Figures/Tables 10

References 42

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富集代数 Enrichment algebra	接种FW（代号1） Inoculated FW （code 1）			接种SS（代号2） Inoculated SS （code 2）
富集代数 Enrichment algebra	碳源A Carbon source A	碳源B Carbon source B	碳源C Carbon source C	碳源A Carbon source A	碳源B Carbon source B	碳源C Carbon source C
第一代（代号-1） First generation （code-1）	1A-1	1B-1	1C-1	2A-1	2B-1	2C-1
第二代（代号-2） Second generation （code-2）	1A-2	1B-2	1C-2	2A-2	2B-2	2C-2
第三代（代号-3） Third generation （code-3）	1A-3	1B-3	1C-3	2A-3	2B-3	2C-3

富集代数 Enrichment algebra	接种FW（代号1） Inoculated FW （code 1）			接种SS（代号2） Inoculated SS （code 2）
富集代数 Enrichment algebra	碳源A Carbon source A	碳源B Carbon source B	碳源C Carbon source C	碳源A Carbon source A	碳源B Carbon source B	碳源C Carbon source C
第一代（代号-1） First generation （code-1）	1A-1	1B-1	1C-1	2A-1	2B-1	2C-1
第二代（代号-2） Second generation （code-2）	1A-2	1B-2	1C-2	2A-2	2B-2	2C-2
第三代（代号-3） Third generation （code-3）	1A-3	1B-3	1C-3	2A-3	2B-3	2C-3

稀释倍数 Dilution ratio	拷贝数 Copy number	C_t值（mean ± SD） Cycle threshold	变异系数 Coefficient of variation （%）
1.00×10^-7	6.53×10³	31.70 ± 0.70	2.21
1.00×10^-6	6.53×10⁴	27.65 ± 0.29	1.05
1.00×10^-5	6.53×10⁵	23.24 ± 0.66	2.84
1.00×10^-4	6.53×10⁶	17.96 ± 0.25	1.40
1.00×10^-3	6.53×10⁷	13.92 ± 0.19	1.36
1.00×10^-2	6.53×10⁸	10.78 ± 0.26	2.39
1.00×10^-1	6.53×10⁹	6.00 ± 0.00	0.00

稀释倍数 Dilution ratio	拷贝数 Copy number	C_t值（mean ± SD） Cycle threshold	变异系数 Coefficient of variation （%）
1.00×10^-7	6.53×10³	31.70 ± 0.70	2.21
1.00×10^-6	6.53×10⁴	27.65 ± 0.29	1.05
1.00×10^-5	6.53×10⁵	23.24 ± 0.66	2.84
1.00×10^-4	6.53×10⁶	17.96 ± 0.25	1.40
1.00×10^-3	6.53×10⁷	13.92 ± 0.19	1.36
1.00×10^-2	6.53×10⁸	10.78 ± 0.26	2.39
1.00×10^-1	6.53×10⁹	6.00 ± 0.00	0.00

Category	Sample	Observed_species	Shannon	Simpson	Pielou	Chao1	ACE	Goods_coverage
产甲烷菌 Methanogens	1A-2	219	4.303	0.911	0.553	291.163	315.584	0.999
	1B-2	204	3.631	0.844	0.483	231.841	254.647	0.999
	1C-2	235	3.709	0.794	0.461	290.724	304.491	0.999
细菌 Bacteria	1A-2	1 644	6.622	0.905	0.620	1 646.686	1 664.078	0.999
	1B-2	1 010	4.234	0.731	0.424	1 028.896	1 062.259	0.997
	1C-2	975	5.925	0.944	0.597	1 046.697	1 079.658	0.995