复合菌群对厌氧消化强化效果及沼气产量影响因素分析

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

摘要/Abstract

摘要：

目的为了获得高效的复合菌群，探究不同生物强化组合对厨余垃圾厌氧消化（anaerobic digestion, AD）产气效果的影响，明确复合菌群提升CH₄产量的作用机制。方法设置添加无菌水（CK）、接种巴氏甲烷八叠球菌（Methanosarcina barkeri）（B）、接种富集的产甲烷菌群（J）、接种复合菌群（M. barkeri与产甲烷菌群按生物量1∶1混合）（BJ）4个处理组，进行序批式AD实验。采用气相色谱及修正的Gompertz模型分析各组的产气特性，随机森林模型分析沼气产量的影响因素。结果所有生物强化处理均延长了产气时间，显著提高了产气速率、累积产气量和CH₄含量（P<0.05），其中BJ组提升效果最显著。BJ组产甲烷潜力最大，其次为J组和B组。温度、底物TS含量、pH值、微量元素（trace elements, TE）强化剂添加量、底物C/N（P<0.01）以及复合菌群接种量（P<0.05）均显著影响沼气产量，其中温度影响最大。局部加权回归（locally weighted regression, LOESS）模型表明，各因素与沼气产量之间存在非线性关系，且每个因素都有其最佳范围。结论复合菌群结合M. barkeri和产甲烷菌群的优势，兼顾功能复杂性与稳定性，同时有效强化上游代谢过程与产甲烷阶段，对AD系统沼气产量和CH₄含量提升效果显著。通过优化生物强化组合和调控关键工艺参数，可有效提升AD性能，推动废弃物资源化利用和能源的可持续发展。

关键词: 产甲烷菌, Methanosarcina barkeri, 生物强化, 微量元素强化, 工艺参数

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

安苗苗, 赵国柱, 徐润, 许菲, 国辉, 李强. 复合菌群对厌氧消化强化效果及沼气产量影响因素分析[J]. 生物技术通报, 2026, 42(2): 89-101.

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.

图/表 11

表1 富集培养基配方

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

表2 厨余垃圾特性

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

图1 序批式AD反应器示意图

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

表3 TE强化剂组成

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

图2 产甲烷菌群的累积产气量及CH4含量不同小写和大写字母表示组间差异显著（P<0.05），下同

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

图3 产甲烷菌群的物种组成与功能特征A：古菌在属水平相对丰度；B：细菌在门水平相对丰度；C：细菌属水平物种组成和系统发育树分析；D：古菌群落功能特征；E：细菌群落功能特征

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

图4 产甲烷菌群的生理特性

Fig. 4 Physiological characteristics of the methanogenic communities

图5 不同生物强化处理组的强化效果A：不同处理组三日沼气产量；B：不同处理组累积沼气产量及CH4含量；C：不同处理组累积CH4产量及修正的Gompertz方程拟合曲线；D：复合菌群的乙酸代谢能力

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

表4 修正的Gompertz模型拟合的相关参数

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

图6 沼气产量影响因素的平均预测重要性*P<0.05， **P<0.01

Fig. 6 Average predictive importance of factors influencing biogas yield

图7 局部加权回归（LOESS）分析沼气产量与显著预测因子之间的关系带有95%置信区间的回归曲线反映了特定因素的预测效果

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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