厌氧消化反应器中同型产乙酸菌产乙酸机制研究
摘要
本文通过对厌氧发酵过程中同型产乙酸作用的研究,提出提高同型产乙酸作用以提高有机质厌氧发酵产乙酸比例的思路;通过对CO_2浓度的变化对有机质厌氧发酵产酸体系中乙酸产率影响的研究,提出CO_2资源化,提高厌氧发酵产酸过程中乙酸比例的方法;通过对富集同型产乙酸菌的富集方法的研究,建立了以甲酸钠作为碳源的混菌富集方法;通过对接种富集前、后的污泥对有机质厌氧发酵产酸结果的比较,验证了提高同型产乙酸作用有利于提高乙酸比例的思路;通过在接种富集后污泥的厌氧发酵产酸体系中通入CO_2的乙酸产率的结果,建立了提高乙酸产率、CO_2减排资源化的方法。全文主要研究内容如下:
1.在中温厌氧发酵产酸体系中,产氢产乙酸菌和同型产乙酸菌具有互营关系。产氢产乙酸菌为同型产乙酸菌提供底物,同型产乙酸菌为产氢产乙酸菌降低氢分压,使产氢产乙酸作用顺利进行。在高氢分压条件下,产氢产乙酸作用不能进行,在CO_2缺失情况下,同型产乙酸作用不能顺利进行。结果表明,改变反应体系中气体成分可以控制两反应的发生状况。以1g/L葡萄糖做模拟废水碳源,接种1g VS/L的种泥,在中温、初始pH为7,抑制产甲烷时,得到同型产乙酸作用对总产乙酸量贡献14.1%,对总VFA产量贡献为6.36%。
2.研究不同顶空CO_2浓度对有机物厌氧发酵体系中底物转化速率、挥发性脂肪酸(VFA)产率及微生物相变化的影响。结果表明,顶空低浓度CO_2有助于葡萄糖底物转化,在16h转化率达到93%以上,顶空高浓度CO_2和对照组在20h时转化率分别为88.3%、87.6%。顶空高浓度CO_2有助于乙酸积累,乙酸产率达8.2mmol/gCOD,分别是对照组和低浓度CO_2组的1.52倍和1.87倍。顶空CO_2浓度与同型产乙酸菌数量正相关,顶空低浓度CO_2组fhs基因为9.83×106拷贝数/mL,高浓度CO_2组fhs基因为5.32×108拷贝数/mL,对照组fhs基因为6.97×107拷贝数/mL。提高顶空CO_2浓度有利于在混和培养环境中富集同型产乙酸菌。
3.在抑制产甲烷的有机质中温厌氧发酵体系中,研究混菌条件下富集同型产乙酸菌的方法。结果表明,甲酸钠作为培养液的碳源可以富集同型产乙酸菌。当甲酸钠浓度为3.25g/L、6.5g/L、13g/L,培养后得到的同型产乙酸菌浓度分别为2.93×109 /mL、6.09×109 /mL和6.08×109 /mL占总菌数的比例为5.68%、12.01%、11.68%。其中甲酸钠为3.25g/L时同型产乙酸菌增长最快,甲酸钠利用率相对较高,为最佳培养液浓度。
4.研究接种富集后的污泥对有机质厌氧发酵产乙酸量的影响。结果表明,接种富集后污泥的乙酸产率是接种未富集污泥的1.38倍。在接种富集污泥的厌氧发酵产酸体系中充入CO_2时,乙酸产率比未充入CO_2提高1.77倍。根据理论核算,接种富集后的污泥并在充入CO_2的方法在经济、环境方面是可行的,在工业方面是具有可操作性的。
This study investigated the contribution of homoacetogenic bacteria during the anaerobic fermentation and proposed the methods for improving the acetate percentage from the organic matters by anaerobic fermentation. By the influence of CO_2 concentration on the acetate yield, a strategy of enhancing acetate production was developed. A novel method for the homoacetogens enrichment was developed based on the formate as carbon source after the study of the enrichment methods. Comparing the production of volatile fatty acids before and after the inoculum of homoacetogen enriched sludge, the idea that homoacetogen is favorable of the acetate production was demonstrated. The detailed contents were as follows:
1. The syntrophic acetogenic bacteria and the homoacetogenic bacteria are belonged to syntrophic relationship in the anaerobic fermentation system. The syntrophic acetogen provides substrate for the homoacetogens and homoacetogen decreased hydrogen partial pressure for the syntrophic acetogens. Under the conditions of high hydrogen partial pressure, the syntrophic acetogenesis can not be occurred while under the condition of absent of CO_2, the homoacetogenesis was not happened. The results showed that change of the composition of the headspace gas can control the acetogenesis process. When the fermentation was run at 1g/L glucose as substrate, 1 1g VS/L seed sludge, pH at 7.0 and the mesophilic conditions, the contribution of the acetogenesis was 14.1% for the total acetate production and 6.36% 2. The effects of CO_2 concentrations in headspace of anaerobic fermentation system on the volatile fatty acids generation, substrate degradation and homoacetogen change were investigated. The results showed that low concentration of headspace CO_2 was helpful for the substrate degradation. The substrate degradation efficiency of low CO_2, high CO_2 and control are over 93% at 16h, 87.6% and 88.3% at 20h, respectively. The high CO_2 concentration improved the acetate accumulation with the conversion yield at 8.2 mmol/g COD at the end of fermentation, which was 1.52 and 1.87 folds of the control and low CO_2 reactors, respectively. There is a positive relationship between CO_2 concentration and the quantity of homoacetogen. The fhs gene copy numbers are 9.83×106/mL, 5.32×108/mL, 6.97×107/mL in the low CO_2, high CO_2 and the control reactors, respectively. The homoacetogen can be enriched from a mixed culture by a high CO_2 concentration in the headspace.
3. The method of homoacetogenic bacteria enrichment was studied in mixed strains. The results showed that the homoacetogenic bacteria could be enriched when the carbon source was sodium formate. When the concentration of sodium formate were 3.25g/L, 6.5g/L and 13g/L, the homoacetogenic bacteria were 2.93×109 /mL, 6.09×109 /mL and 6.08×109 /mL, respectively, which were 5.68%, 12.01% and 11.68% of the total bacteria. The growth rate of homoacetogenic bacteria and utilization of sodium formate were the fastest during the enrichment period when the sodium formate was 3.25g/L.
4. The effect of inoculum with homoacetogenic enriched sludge on the acetate yield was investigated. The results showed that the yield of acetate in the reactors which homoacetogen enriched sludge was 1.38 folds higher than the reactors without homoacetogen enriched sludge. The yield of acetate which filled with CO_2 in the reactors with homoacetogen enriched sludge was 1.77 folds higher than the CO_2 absent group. Based on the theoretic calculation, the inoculum with homoacetogen in coupled with CO_2 filling is a promising and feasible strategy for the pollution control.
1.在中温厌氧发酵产酸体系中,产氢产乙酸菌和同型产乙酸菌具有互营关系。产氢产乙酸菌为同型产乙酸菌提供底物,同型产乙酸菌为产氢产乙酸菌降低氢分压,使产氢产乙酸作用顺利进行。在高氢分压条件下,产氢产乙酸作用不能进行,在CO_2缺失情况下,同型产乙酸作用不能顺利进行。结果表明,改变反应体系中气体成分可以控制两反应的发生状况。以1g/L葡萄糖做模拟废水碳源,接种1g VS/L的种泥,在中温、初始pH为7,抑制产甲烷时,得到同型产乙酸作用对总产乙酸量贡献14.1%,对总VFA产量贡献为6.36%。
2.研究不同顶空CO_2浓度对有机物厌氧发酵体系中底物转化速率、挥发性脂肪酸(VFA)产率及微生物相变化的影响。结果表明,顶空低浓度CO_2有助于葡萄糖底物转化,在16h转化率达到93%以上,顶空高浓度CO_2和对照组在20h时转化率分别为88.3%、87.6%。顶空高浓度CO_2有助于乙酸积累,乙酸产率达8.2mmol/gCOD,分别是对照组和低浓度CO_2组的1.52倍和1.87倍。顶空CO_2浓度与同型产乙酸菌数量正相关,顶空低浓度CO_2组fhs基因为9.83×106拷贝数/mL,高浓度CO_2组fhs基因为5.32×108拷贝数/mL,对照组fhs基因为6.97×107拷贝数/mL。提高顶空CO_2浓度有利于在混和培养环境中富集同型产乙酸菌。
3.在抑制产甲烷的有机质中温厌氧发酵体系中,研究混菌条件下富集同型产乙酸菌的方法。结果表明,甲酸钠作为培养液的碳源可以富集同型产乙酸菌。当甲酸钠浓度为3.25g/L、6.5g/L、13g/L,培养后得到的同型产乙酸菌浓度分别为2.93×109 /mL、6.09×109 /mL和6.08×109 /mL占总菌数的比例为5.68%、12.01%、11.68%。其中甲酸钠为3.25g/L时同型产乙酸菌增长最快,甲酸钠利用率相对较高,为最佳培养液浓度。
4.研究接种富集后的污泥对有机质厌氧发酵产乙酸量的影响。结果表明,接种富集后污泥的乙酸产率是接种未富集污泥的1.38倍。在接种富集污泥的厌氧发酵产酸体系中充入CO_2时,乙酸产率比未充入CO_2提高1.77倍。根据理论核算,接种富集后的污泥并在充入CO_2的方法在经济、环境方面是可行的,在工业方面是具有可操作性的。
This study investigated the contribution of homoacetogenic bacteria during the anaerobic fermentation and proposed the methods for improving the acetate percentage from the organic matters by anaerobic fermentation. By the influence of CO_2 concentration on the acetate yield, a strategy of enhancing acetate production was developed. A novel method for the homoacetogens enrichment was developed based on the formate as carbon source after the study of the enrichment methods. Comparing the production of volatile fatty acids before and after the inoculum of homoacetogen enriched sludge, the idea that homoacetogen is favorable of the acetate production was demonstrated. The detailed contents were as follows:
1. The syntrophic acetogenic bacteria and the homoacetogenic bacteria are belonged to syntrophic relationship in the anaerobic fermentation system. The syntrophic acetogen provides substrate for the homoacetogens and homoacetogen decreased hydrogen partial pressure for the syntrophic acetogens. Under the conditions of high hydrogen partial pressure, the syntrophic acetogenesis can not be occurred while under the condition of absent of CO_2, the homoacetogenesis was not happened. The results showed that change of the composition of the headspace gas can control the acetogenesis process. When the fermentation was run at 1g/L glucose as substrate, 1 1g VS/L seed sludge, pH at 7.0 and the mesophilic conditions, the contribution of the acetogenesis was 14.1% for the total acetate production and 6.36% 2. The effects of CO_2 concentrations in headspace of anaerobic fermentation system on the volatile fatty acids generation, substrate degradation and homoacetogen change were investigated. The results showed that low concentration of headspace CO_2 was helpful for the substrate degradation. The substrate degradation efficiency of low CO_2, high CO_2 and control are over 93% at 16h, 87.6% and 88.3% at 20h, respectively. The high CO_2 concentration improved the acetate accumulation with the conversion yield at 8.2 mmol/g COD at the end of fermentation, which was 1.52 and 1.87 folds of the control and low CO_2 reactors, respectively. There is a positive relationship between CO_2 concentration and the quantity of homoacetogen. The fhs gene copy numbers are 9.83×106/mL, 5.32×108/mL, 6.97×107/mL in the low CO_2, high CO_2 and the control reactors, respectively. The homoacetogen can be enriched from a mixed culture by a high CO_2 concentration in the headspace.
3. The method of homoacetogenic bacteria enrichment was studied in mixed strains. The results showed that the homoacetogenic bacteria could be enriched when the carbon source was sodium formate. When the concentration of sodium formate were 3.25g/L, 6.5g/L and 13g/L, the homoacetogenic bacteria were 2.93×109 /mL, 6.09×109 /mL and 6.08×109 /mL, respectively, which were 5.68%, 12.01% and 11.68% of the total bacteria. The growth rate of homoacetogenic bacteria and utilization of sodium formate were the fastest during the enrichment period when the sodium formate was 3.25g/L.
4. The effect of inoculum with homoacetogenic enriched sludge on the acetate yield was investigated. The results showed that the yield of acetate in the reactors which homoacetogen enriched sludge was 1.38 folds higher than the reactors without homoacetogen enriched sludge. The yield of acetate which filled with CO_2 in the reactors with homoacetogen enriched sludge was 1.77 folds higher than the CO_2 absent group. Based on the theoretic calculation, the inoculum with homoacetogen in coupled with CO_2 filling is a promising and feasible strategy for the pollution control.
引文
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