吲哚降解菌及其除臭功能研究

发布时间：2021-03-28 09:37

　　吲哚类化合物引起的环境污染日益严重。在动物养殖中,来源于L-色氨酸无氧代谢产生的粪臭素（3-甲基吲哚,3MI）等吲哚类化合物是养殖场臭气的主要成分,也是造成养殖场及其周边环境恶化的主要因素。如何有效治理吲哚污染是养殖业可持续发展所面临的难题。本研究使用富集培养法从家禽粪便中分离出能够利3MI和吲哚的可培养细菌,在进行生化和分子鉴定的基础上研究了其生化特性、培养条件、除臭功能及其机理,为吲哚生物除臭饲料添加剂的研制奠定基础。本研究分离获得能够高效降解粪臭素的细菌2株,通过使用Biolog GEN III MicroPlate评估所分离菌株的生化特征并依据16S rDNA基因测序结果,将其鉴定为不动杆菌NTA1-2A（A.Toweneri）和不动杆菌TAT1-6A（A.guillouiae）。通过单因素实验确定了降解粪臭素（3MI）、吲哚的最佳温度和pH值,评估对粪臭素和吲哚的初始浓度的耐受性以及初始浓度对降解效果的影响。结果表明,菌株NTA1-2A和TAT1-6A降解3MI的最佳温度和pH分别为31℃和6,两菌株可有效降解初始浓度范围为65²00mg/L的3MI;当... 

【文章来源】：中国农业科学院北京市

【文章页数】：107 页

【学位级别】：博士

【文章目录】：
摘要
abstract
LIST OF ACRONYMS
CHAPTER1 INTRODUCTION
    1.1 GENERAL INTRODUCTION
CHAPTER2 LITERATURE REVIEW
    2.1 STUDY BACKGROUND
    2.2 FORMATION OF INDOLE AND SKATOLE
    2.3 3-METHYLINDOLE（SKATOLE）
        2.3.1 Effects of3-methylindole
        2.3.2 Bacterial degradation of3-methylindole
    2.4 MICROBIAL DEGRADATION OF INDOLE
        2.4.1 Anaerobic bacteria degradation of indole
        2.4.2 Aerobic degradation of indole
        2.4.3 Bacterial cometabolism of indole
        2.4.4 Fungal degradation of indole
    2.5 THE CURRENT STUDY
        2.5.1 Aims and objectives of current study
        2.5.2 Purpose and significance of the study
CHAPTER3 ISOLATION AND MOLECULAR CHARACTERIZATION OF3-METHYLINDOLEDEGRADING BACTERIA FROM POULTRY MANURE
    3.1 INTRODUCTION
    3.2 MATERIALS AND METHODS
        3.2.1 Bacteria growth medium
        3.2.2 Identification of the selected strains NTA1-2A and TAT-6A
        3.2.3 Assessing3MI degradation
        3.2.4 Single factors affecting3MI degradation
        3.2.5 Effect of3MI initial concentrations
        3.2.6 Enzyme activity test
        3.2.7 Poultry manure fermentation experiment
        3.2.8 Sample preparation for HPLC analysis
    3.3 STATISTICAL ANALYSIS
    3.4 RESULTS
        3.4.1 Identification of strains NTA1-2A and TAT1-6A
        3.4.2 Effect of temperature and pH on growth and3MI Degradation
        3.4.3 3MI degradation at different initial concentrations
        3.4.4 3MI degradation efficiency of supernatant
        3.4.5 3MI removal from poultry manure
    3.5 DISCUSSION
    3.6 CONCLUSION
CHAPTER4 DEGRADATION OF INDOLE USING A.TOWENERI NTA1-2A AND A.GUILLOUIAE TAT1-6A BACTERIAL STRAINS
    4.1 INTRODUCTION
    4.2 MATERIALS AND METHODS
        4.2.1 Source of microorganisms
        4.2.2 Medium conditions and bacteria growth
        4.2.3 Indole degradation by NTA1-2A and TAT1-6A
        4.2.4 Factors affecting Indole degradation
        4.2.5 Enzyme activity test
        4.2.6 Poultry manure fermentation
        4.2.7 Sample preparation for HPLC analysis
    4.3 STATISTICAL ANALYSIS
    4.4 RESULTS
        4.4.1 Indole degradation efficiency
        4.4.2 Indole degradation at different initial concentrations
        4.4.3 Factors affecting Indole degradation
        4.4.4 Effect of indole on bacterial growth
        4.4.5 Indole degradation efficiency of supernatant
        4.4.6 Indole removal from poultry manure
    4.5 DISCUSSION
    4.6 CONCLUSION
CHAPTER5 ANALYSIS OF3-METHYLINDOLE AND INDOLE DEGRADATIONMETABOLITES
    5.1 INTRODUCTION
        5.1.1 3-methylindole degradation metabolites
        5.1.2 Indole degradation metabolites
    5.2 MATERIALS AND METHODS
        5.2.1 Source of microorganisms and Medium conditions
        5.2.2 Sample preparation and LC– MS/MS analysis
        5.2.3 Data analysis
        5.2.4 Protein identification by SDS-PAGE
    5.3 RESULTS
        5.3.1 3-methylindole degradation metabolites
        5.3.2 Indole degradation metabolites
        5.3.3 Metabolic enzymes
    5.4 DISCUSSION
    5.5 CONCLUSIONS
CHAPTER6 OVERALL CONCLUSIONS AND RECOMMENDATIONS
    6.1 CONCLUSIONS
    6.2 RECOMMENDATIONS
CHAPTER7 REFERENCES
ACKNOWLEDGMENTS
AUTHOR RESUME
APPENDICES



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