正交定向调控VFAs奇偶数比率对合成PHAs的影响
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摘要
为了定向调控挥发性脂肪酸(VFAs)中奇偶数碳比率,在合成聚羟基链烷酯(PHAs)过程中改变PHB和PHV的比率,从而改变可生物降解塑料的内在性能。以淘米水为单一发酵对象,将pH值、温度、发酵天数作为影响因素,通过L9(33)正交试验观察VFAs中奇偶数脂肪酸变化,和污泥中聚磷菌合成PHAs的变化规律。结果表明当温度为35℃,pH值为8,第7天时奇偶数碳比率约为1∶1;当温度为25℃,p H值为8,第9天时奇偶数碳比率约为1∶2;当温度为45℃,pH值为6,第7天时奇偶数碳比率约为1∶3。将这3种不同比率的组合等量投加至污泥中,PHAs含量有显著增加,其中发酵液奇偶数碳比率为1∶1时PHAs积累量最高,为80.69 mg/L。研究表明pH值对偶数脂肪酸影响最大,发酵天数对奇数脂肪酸影响最大。而且发酵液比化学碳源更有利于PHAs的合成,使厌氧阶段PHB的质量分数显著增加,好氧阶段PHV的质量分数显著增加。
In order to directionally regulate odd-to-even carbon-number ratio of volatile fatty acids(VFAs) and to alter the poly-β-hydroxybutyrate-to-polyhydroxyvalerate(PHB/PHV) ratio of polyhydroxy chain alkyl ester(PHAs) for the purpose of changing the intrinsic properties of biodegradable plastics,the rice-washing water was used as a single fermentation study object,and through a L9(33) orthogonal test with temperature,pH value and fermentation time as influencing factors,an experiment was conducted to investigate the variation of odd-to-even carbon-number ratio of VFAs,and the PHAs synthesis of polyphosphate accumulating organism(PAO) in the sludge.The study suggested that on condition that pH value was 8,temperature 35 ℃ and the fermentation time seven days,the odd-to-even carbon-number ratio of VFAs was 1∶1;in the case where pH value was 8,temperature 25 ℃,the fermentation time nine days,the ratio was 1∶2;and when pH value was 6,temperature 45 ℃,the fermentation time being seven days,the ratio was 1∶3.Thereafter,to put equal amounts of fermentation broth of those three different ratios into the acclimated sludge,which resulted in a significant increase of PHAs content,and when odd-to-even carbon-number ratio of VFAs was 1∶1,PHAs accumulated attained the highest 80.69 mg/L.In general,pH value affected the even-numbered fatty acids most,while the number of fermentation days has the greatest effect on oddnumbered fatty acids.The fermentation broth is more conducive to PHAs accumulation than the chemical carbon source;therefore,the mass fraction of PHB in the anaerobic phase was remarkably increased,as the mass fraction of PHV in the aerobic phase.
In order to directionally regulate odd-to-even carbon-number ratio of volatile fatty acids(VFAs) and to alter the poly-β-hydroxybutyrate-to-polyhydroxyvalerate(PHB/PHV) ratio of polyhydroxy chain alkyl ester(PHAs) for the purpose of changing the intrinsic properties of biodegradable plastics,the rice-washing water was used as a single fermentation study object,and through a L9(33) orthogonal test with temperature,pH value and fermentation time as influencing factors,an experiment was conducted to investigate the variation of odd-to-even carbon-number ratio of VFAs,and the PHAs synthesis of polyphosphate accumulating organism(PAO) in the sludge.The study suggested that on condition that pH value was 8,temperature 35 ℃ and the fermentation time seven days,the odd-to-even carbon-number ratio of VFAs was 1∶1;in the case where pH value was 8,temperature 25 ℃,the fermentation time nine days,the ratio was 1∶2;and when pH value was 6,temperature 45 ℃,the fermentation time being seven days,the ratio was 1∶3.Thereafter,to put equal amounts of fermentation broth of those three different ratios into the acclimated sludge,which resulted in a significant increase of PHAs content,and when odd-to-even carbon-number ratio of VFAs was 1∶1,PHAs accumulated attained the highest 80.69 mg/L.In general,pH value affected the even-numbered fatty acids most,while the number of fermentation days has the greatest effect on oddnumbered fatty acids.The fermentation broth is more conducive to PHAs accumulation than the chemical carbon source;therefore,the mass fraction of PHB in the anaerobic phase was remarkably increased,as the mass fraction of PHV in the aerobic phase.
引文
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[10]王权,宫常修,蒋建国,等.NaCl对餐厨垃圾厌氧发酵产VFA浓度及组分的影响[J].中国环境科学,2014,34(12):3127-3132.Wang Quan,Gong Changxiu,Jiang Jianguo,et al.Effect of NaCl content on VFA concentration and composition during anaerobic fermentation of kitchen waste[J].China Environmental Science,2014,34(12):3127-3132.
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[12]宋雪健.淘米水物质成分分析[J].科技创新与应用,2015(17):41-41.
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[14]Chaikasem S,Abeynayaka A,Visvanathan C.Effect of polyvinyl alcohol hydrogel as a biocarrier on volatile fatty acids production of a two-stage thermophilic anaerobic membrane bioreactor[J].Bioresource Technology,2014,168(3):100-105.
[15]Moreno andrade I,Carrillo Reyes J,Santiago S G,et al.Biohydrogen from food waste in a discontinuous process:effect of HRT and microbial community analysis[J].International Journal of Hydrogen Energy,2015,40(48):17246-17252.
[16]Page L H,Ni J Q,Zhang H,et al.Reduction of volatile fatty acids and odor offensiveness by anaerobic digestion and solid separation of dairy manure during manure storage[J].Journal of Environmental Management,2015,152:91-98.
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[18]Arreola vargas J,Flores larios A,GonzalezáLvarez V,et al.Single and two-stage anaerobic digestion for hydrogen and methane production from acid and enzymatic hydrolysates of Agave tequilana,bagasse[J].International Journal of Hydrogen Energy,2016,41(2):897-904.
[19]李定龙,戴肖云,赵宋敏,等.pH对厨余垃圾厌氧产酸的影响[J].环境科学与技术,2011,34(4):125-128.Li Dinglong,Dai Xiaoyun,Zhao Songmin,et al.Influence of pH on acidity during anaerobic fermentation of kitchen waste[J].Environmental Science&Technology,2011,34(4):125-128.
[20]Han S K,Kim S H,Shin H S.UASB treatment of wastewater with VFA and alcohol generated during hydrogen fermentation of food waste[J].Process Biochemistry,2005,40(8):2897-2905.
[21]Rikame S S,Mungray A A,Mungray A K.Electricity generation from acidogenic food waste leachate using dual chamber mediator less microbial fuel cell[J].International Biodeterioration&Biodegradation,2012,75(6):131-137.
[22]吕凡,何品晶,邵立明,等.p H值对易腐有机垃圾厌氧发酵产物分布的影响[J].环境科学,2006,27(5):991-997.Lyu Fan,He Pinjing,Shao Liming,et al.Effect of pH value fermentation pathways of biodegradable organic waste[J].Environmental Science,2006,27(5):991-997.
[23]李阳,周涛,陈善平,等.温度对接种酵母菌和醋酸菌餐厨垃圾微氧发酵产乙酸的影响[J].中国环境科学,2016,36(1):175-180.Li Yang,Zhou Tao,Chen Shanping,et al.Effects of temperature on the production of acetic acid from food wastes by yeast and acetic acid bacteria during micro-aerobic fermentation[J].China Environmental Science,2016,36(1):175-180.
[24]Lim S J,Kim B J,Jeong C M,et al.Anaerobic organic acid production of food waste in once-a-day feeding and drawing-off bioreactor[J].Bioresour Technol,2008,99(16):7866-7874.
[25]国家环境保护总局《水和废水监测分析方法》编委会.水和废水监测分析方法[M].第四版.北京:中国环境科学出版社,2002.
[26]Li X,Chen Y,Zhao S,et al.Lactic acid accumulation from sludge and food waste to improve the yield of propionic acid-enriched VFA[J].Biochemical Engineering Journal,2014,84(6):28-35.
[27]蔡萌萌.剩余活性污泥中的微生物利用实际废液合成聚羟基烷酸酯[D].哈尔滨:哈尔滨工业大学,2009.Cai Mengmeng.Polyalkanoates Synthesis from Real Waste by Microorganisms in Excess Sludge[D].Harbin:Harbin Institute of Technology,2009.
[28]周爱娟.预处理促进剩余污泥发酵制取短链脂肪酸的效能及机制研究[D].哈尔滨:哈尔滨工业大学,2014.Zhou Aijuan.Short Chain Fatty Acids Production from Waste Activated Sludge Fermentation Stimulated by Pretreatments[D].Harbin:Harbin Institute of Technology,2014.
[29]Elefsiniotis P,Wareham D G.Utilization patterns of volatile fatty acids in the denitrification reaction[J].Enzyme&Microbial Technology,2007,41(2):92-97.
[30]余晓琴.餐厨垃圾厌氧发酵产酸优化及蛋白质组分的产酸特性研究[D].杭州:浙江工商大学,2017.Yu Xiaoqin.Optimization of Acidogenic Fermentation from Food Waste and Fermentative Characteristics of Proteinrich Substrates[D].Hangzhou:Zhejiang Gongshang University,2017.
[31]黄龙.混和菌群合成PHA工艺影响因子与优化策略研究[D].哈尔滨:哈尔滨工业大学,2013.Huang Long.Research on Influence Factors and Optimized Strategies of Mixed Cultures PHA Production Process[D].Harbin:Harbin Institute of Technology,2013.
[32]Serafim L S,Lemos P C,Albuquerque M G E,et al.Strategies for PHA production by mixed cultures and renewable waste materials[J].Applied Microbiology&Biotechnology,2008,81(4):615-628.
[2]Volova T,Shishatskaya E,Sevastianov V,et al.Results of biomedical investigations of PHB and PHB/PHV fibers[J].Biochemical Engineering Journal,2003,16(2):125-133.
[3]Brigham C J,Zhila N,Shishatskaya E,et al.Manipulation of Ralstonia eutropha Carbon Storage Pathways to Produce Useful Bio-based Products:Reprogramming Microbial Metabolic Pathways[M].Springer Netherlands,2012.
[4]Albuquerquel M G E,Martino V,Pollet E,et al.Mixed culture polyhydroxyalkanoate(PHA)production from volatile fatty acid(VFA)-rich streams:effect of substrate composition and feeding regime on PHA productivity,composition and properties[J].Journal of Biotechnology,2011,151(1):66.
[5]Fradinho J C,Oehmen A,Reis M A.Photosynthetic mixed culture polyhydroxyalkanoate(PHA)production from individual and mixed volatile fatty acids(VFAs):substrate preferences and co-substrate uptake[J].Journal of Biotechnology,2014,185:19.
[6]Suriyamongkol P,Weselake R,Narine S,et al.Biotechnological approaches for the production of polyhydroxyalkanoates in microorganisms and plants-a review[J].Biotechnology Advances,2007,25(2):148-175.
[7]Oliveira F C,Dias M L,Castilho L R,et al.Characterization of poly(3-hydroxybutyrate)produced by Cupriavidus necator in solid-state fermentation[J].Bioresource Technology,2007,98(3):633-638.
[8]Zhang M,Wu H,Chen H.Coupling of polyhydroxyalkanoate production with volatile fatty acid from food wastes and excess sludge[J].Process Safety&Environmental Protection Transactions of the I,2014,92(2):171-178.
[9]熊丹丹.废弃碳源定向产酸过程调控与合成PHA的应用研究[D].哈尔滨:哈尔滨工业大学,2016.Xiong Dandan.Research on Process of Direction Acids Regulation and PHA Production Using Waste Carbon Resource[D].Harbin:Harbin Institute of Technology,2016.
[10]王权,宫常修,蒋建国,等.NaCl对餐厨垃圾厌氧发酵产VFA浓度及组分的影响[J].中国环境科学,2014,34(12):3127-3132.Wang Quan,Gong Changxiu,Jiang Jianguo,et al.Effect of NaCl content on VFA concentration and composition during anaerobic fermentation of kitchen waste[J].China Environmental Science,2014,34(12):3127-3132.
[11]付云霞.城市生活垃圾单基质厌氧消化研究[D].成都:西南交通大学,2007.
[12]宋雪健.淘米水物质成分分析[J].科技创新与应用,2015(17):41-41.
[13]Dahiya S,Sarkar O,Swamy Y V,et al.Acidogenic fermentation of food waste for volatile fatty acid production with co-generation of biohydrogen[J].Bioresource Technology,2015,182:103-113.
[14]Chaikasem S,Abeynayaka A,Visvanathan C.Effect of polyvinyl alcohol hydrogel as a biocarrier on volatile fatty acids production of a two-stage thermophilic anaerobic membrane bioreactor[J].Bioresource Technology,2014,168(3):100-105.
[15]Moreno andrade I,Carrillo Reyes J,Santiago S G,et al.Biohydrogen from food waste in a discontinuous process:effect of HRT and microbial community analysis[J].International Journal of Hydrogen Energy,2015,40(48):17246-17252.
[16]Page L H,Ni J Q,Zhang H,et al.Reduction of volatile fatty acids and odor offensiveness by anaerobic digestion and solid separation of dairy manure during manure storage[J].Journal of Environmental Management,2015,152:91-98.
[17]张玉静,蒋建国,王佳明.p H值对餐厨垃圾厌氧发酵产挥发性脂肪酸的影响[J].中国环境科学,2013,33(4):680-684.Zhang Yujing,Jiang Jianguo,Wang Jiaming.Effect of pHvalue on VFA concentration and composition during anaerobic fermentation of kitchen waste[J].China Environmental Science,2013,33(4):680-684.
[18]Arreola vargas J,Flores larios A,GonzalezáLvarez V,et al.Single and two-stage anaerobic digestion for hydrogen and methane production from acid and enzymatic hydrolysates of Agave tequilana,bagasse[J].International Journal of Hydrogen Energy,2016,41(2):897-904.
[19]李定龙,戴肖云,赵宋敏,等.pH对厨余垃圾厌氧产酸的影响[J].环境科学与技术,2011,34(4):125-128.Li Dinglong,Dai Xiaoyun,Zhao Songmin,et al.Influence of pH on acidity during anaerobic fermentation of kitchen waste[J].Environmental Science&Technology,2011,34(4):125-128.
[20]Han S K,Kim S H,Shin H S.UASB treatment of wastewater with VFA and alcohol generated during hydrogen fermentation of food waste[J].Process Biochemistry,2005,40(8):2897-2905.
[21]Rikame S S,Mungray A A,Mungray A K.Electricity generation from acidogenic food waste leachate using dual chamber mediator less microbial fuel cell[J].International Biodeterioration&Biodegradation,2012,75(6):131-137.
[22]吕凡,何品晶,邵立明,等.p H值对易腐有机垃圾厌氧发酵产物分布的影响[J].环境科学,2006,27(5):991-997.Lyu Fan,He Pinjing,Shao Liming,et al.Effect of pH value fermentation pathways of biodegradable organic waste[J].Environmental Science,2006,27(5):991-997.
[23]李阳,周涛,陈善平,等.温度对接种酵母菌和醋酸菌餐厨垃圾微氧发酵产乙酸的影响[J].中国环境科学,2016,36(1):175-180.Li Yang,Zhou Tao,Chen Shanping,et al.Effects of temperature on the production of acetic acid from food wastes by yeast and acetic acid bacteria during micro-aerobic fermentation[J].China Environmental Science,2016,36(1):175-180.
[24]Lim S J,Kim B J,Jeong C M,et al.Anaerobic organic acid production of food waste in once-a-day feeding and drawing-off bioreactor[J].Bioresour Technol,2008,99(16):7866-7874.
[25]国家环境保护总局《水和废水监测分析方法》编委会.水和废水监测分析方法[M].第四版.北京:中国环境科学出版社,2002.
[26]Li X,Chen Y,Zhao S,et al.Lactic acid accumulation from sludge and food waste to improve the yield of propionic acid-enriched VFA[J].Biochemical Engineering Journal,2014,84(6):28-35.
[27]蔡萌萌.剩余活性污泥中的微生物利用实际废液合成聚羟基烷酸酯[D].哈尔滨:哈尔滨工业大学,2009.Cai Mengmeng.Polyalkanoates Synthesis from Real Waste by Microorganisms in Excess Sludge[D].Harbin:Harbin Institute of Technology,2009.
[28]周爱娟.预处理促进剩余污泥发酵制取短链脂肪酸的效能及机制研究[D].哈尔滨:哈尔滨工业大学,2014.Zhou Aijuan.Short Chain Fatty Acids Production from Waste Activated Sludge Fermentation Stimulated by Pretreatments[D].Harbin:Harbin Institute of Technology,2014.
[29]Elefsiniotis P,Wareham D G.Utilization patterns of volatile fatty acids in the denitrification reaction[J].Enzyme&Microbial Technology,2007,41(2):92-97.
[30]余晓琴.餐厨垃圾厌氧发酵产酸优化及蛋白质组分的产酸特性研究[D].杭州:浙江工商大学,2017.Yu Xiaoqin.Optimization of Acidogenic Fermentation from Food Waste and Fermentative Characteristics of Proteinrich Substrates[D].Hangzhou:Zhejiang Gongshang University,2017.
[31]黄龙.混和菌群合成PHA工艺影响因子与优化策略研究[D].哈尔滨:哈尔滨工业大学,2013.Huang Long.Research on Influence Factors and Optimized Strategies of Mixed Cultures PHA Production Process[D].Harbin:Harbin Institute of Technology,2013.
[32]Serafim L S,Lemos P C,Albuquerque M G E,et al.Strategies for PHA production by mixed cultures and renewable waste materials[J].Applied Microbiology&Biotechnology,2008,81(4):615-628.