Bioaugmentation Effect of Compound Microbial Consortium on Anaerobic Digestion and Factors Influencing Biogas Yield

doi:10.13560/j.cnki.biotech.bull.1985.2025-0169

Abstract

Abstract:

Objective To obtain an efficient compound microbial consortium, the effects of different bioaugmentation treatments on biogas yield in food waste anaerobic digestion (AD) were investigated to elucidate the mechanisms by which the compound microbial consortium enhances CH₄ yield. Method Four treatments were set up for the sequential batch AD experiment: addition of sterile water (CK), inoculation with Methanosarcina barkeri (B), inoculation with enriched methanogenic community (J), and inoculation with constructed microbial consortium comprising M. barkeri and methanogenic community in a 1∶1 based on biomass (BJ). The characteristics of biogas production were analyzed using gas chromatography and the modified Gompertz model. Additionally, a random forest model was utilized to identify the key factors influencing biogas yield. Result The results showed that all bioaugmentation treatments extended the biogas production period and significantly enhanced the biogas production rate, cumulative biogas yield, and CH₄ content (P<0.05). Notably, the BJ group showed the most significant improvement. BJ group presented the highest methanogenic potential, followed by J and B groups. Temperature, substrate TS content, pH value, trace elements (TE) concentration, substrate C/N ratio (P<0.01), and inoculation amount of the compound microbial consortium all had significant effects on biogas yield (P<0.05), with temperature exerting the most substantial impact. The locally weighted regression (LOESS) model showed a nonlinear relationship between each factor and biogas yield, with each factor having its optimal range. Conclusion The compound microbial consortium integrated the advantages of the methanogenic community and M. barkeri, balancing functional complexity and stability. This effectively enhanced upstream metabolic processes and methanogenesis, significantly increasing biogas yield and CH₄ content in the AD system. By optimizing the bioaugmentation combination and regulating key process parameters, the performance of AD can be effectively improved, promoting waste resource utilization and the sustainable development of energy.

Key words: methanogens, Methanosarcina barkeri, bioaugmentation, trace elements supplementation, process parameter

AN Miao-miao, ZHAO Guo-zhu, XU Run, XU Fei, GUO Hui, LI Qiang. Bioaugmentation Effect of Compound Microbial Consortium on Anaerobic Digestion and Factors Influencing Biogas Yield[J]. Biotechnology Bulletin, 2026, 42(2): 89-101.

Figures/Tables 11

Table 1 Formula of enrichment culture medium

成分Component	含量Content	成分Component	含量Content
NH₄Cl	1 g	FeSO₄·7H₂O溶液	2 mL （0.1 g/mL）
MgSO₄·7H₂O	0.5 g	Cysteine-HCl	0.3 g
K₂HPO₄	0.4 g	CaCl₂·2H₂O	0.2 g
KH₂PO₄	0.2 g	Na₂S·9H₂O	25.0 mL （0.012 g/mL）
HCOONa	5.0 g	NaHCO₃	20 mL （0.1 g/mL）
CH₃COONa	5.0 g	TE溶液^［19］	1.0 mL
50% CH₃OH	4 mL	维生素溶液^［20］	10.0 mL
Yeast extract	1.0 g	氨苄青霉素	4.0 mL （0.1 g/mL）
Tryptone	1.0 g	刃天青钠溶液	0.5 mL （0.001 g/mL）
NaCl	2.3 g	蒸馏水	至1 000 mL

Table 2 Characteristics of food waste

参数 Parameter	Mean ± SD （%）
TS	25.94 ± 0.07
VS	24.21 ± 0.08
VS/TS	93.33 ± 0.01
C	47.91 ± 0.79
H	6.31 ± 0.03
N	3.70 ± 0.07
S	0.36 ± 0.05
C/N	12.96 ± 0.03

Fig. 1 Diagram of anaerobic digestion (AD) reactor in batch mode

Table 3 Trace elements enhancer composition

成分 Component	含量 Content（g/L）	成分 Component	含量 Content（g/L）
NH₄Cl	20.0	CoCl₂·6H₂O	0.2
K₂HPO₄	4.0	NiCl₂·6H₂O	0.2
KH₂PO₄	2.0	CaO	0.2
FeSO₄·7H₂O	2.0	Na₂MoO₄·2H₂O	0.2
MgCl₂	2.0	CuSO₄·5H₂O	0.2
KCl	2.0	ZnCl₂	0.2

Fig. 2 Cumulative gas yield and methane content of the methanogenic communityDifferent lowercase and uppercase letters indicate significant differences between groups, the same below

Fig. 3 Species composition and functional characteristics of the methanogenic communityA: Distribution of archaeal at the genus level. B: Distribution of bacterial at the phylum level. C: Composition and cladogram analysis of bacteria at the genus level. D: Function characteristics of archaeal community. E: Function characteristics of bacterial community

Fig. 4 Physiological characteristics of the methanogenic communities

Fig. 5 Enhancement effects of different bioaugmentation treatment groupsA: Three-day biogas yield of different groups. B: Cumulative biogas yield and methane content of different groups. C: Cumulative methane yield and the modified Gompertz equation fitting curve of different groups. D: Acetic acid metabolic capacity of compound microbial consortium

Table 4 Fitting parameters obtained from the modified Gompertz model

组别Group	最大产甲烷潜力Maximum methane production potential M_max （mL）	最大产甲烷速率Maximum methane production rate R_max （mL/d）	延滞期Lag phase λ （d）	相关系数Correlation coefficient R²
CK	2.839 ± 0.042	0.327 ± 0.026	3.917 ± 0.363	0.995 5
B	195.108 ± 3.507	21.256 ± 1.820	5.499 ± 0.408	0.995 0
J	327.591 ± 6.058	28.595 ± 1.989	5.124 ± 0.400	0.996 1
BJ	972.266 ± 43.703	66.523 ± 7.121	6.698 ± 0.732	0.989 8

Fig. 6 Average predictive importance of factors influencing biogas yield

Fig. 7 Relationships between biogas yield and the significant predictors by locally weighted regression analysisThe regression curve with a 95% confidence interval reflects the predictive effect of specific factors

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