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糖巨肽蛋白源DPP-4抑制肽的制备、鉴定及抑制机理研究

摘要第6-7页
ABSTRACT第7页
Abbreviation第15-16页
CHAPTER 1 Background and objectives第16-25页
    1.1 General introduction第16-17页
    1.2 Milk Proteins第17-20页
        1.2.1 Overview of bioactive peptides第18页
        1.2.2 Milk-derived bioactive peptides第18页
        1.2.3 Enzymatic hydrolysis by digestive enzymes第18-19页
        1.2.4 Microbial fermentation第19-20页
    1.3 Bioactivities from peptides and health significance第20-23页
        1.3.1 Antidiabetic functionalities of milk protein-derived bioactive peptides第21页
        1.3.2 Effect of milk protein on insulin secretion第21页
        1.3.3 Antidiabetic applications of bioactive peptides第21页
        1.3.4 In vitro antidiabetic effect of peptides第21-23页
    1.4 Objectives of the study第23-25页
        1.4.1 The Specific Objectives of this Research are:第23-25页
CHAPTER 2 Effects of microwave and ultrasound pretreatments on preparation of Glycomacropeptide withtrypsin as a source DPP-IV inhibitory peptide第25-36页
    2.1 Introduction第25-26页
    2.2 Materials and methods第26-30页
        2.2.1 Materials and reagents第26页
        2.2.2 Instruments equipments第26页
        2.2.3 Determination of protein content第26页
        2.2.4 Protease activity assay第26页
        2.2.5 Trichloroacetic acid precipitation -Determination of short peptide content by forinolmethod第26-28页
        2.2.6 Determination of the degree of hydrolysis of enzymatically digested samples by the o-Phthalaldehyde method (OPA method)第28页
        2.2.7 Microwave and ultrasonic pretreatments第28页
        2.2.8 Enzyme hydrolysis and degree of hydrolysis第28-29页
        2.2.9 Assay of the DPP-IV-Inhibitory Activity第29页
        2.2.10 Sttistical analysis第29-30页
    2.3 Results and discussions第30-34页
        2.3.1 Protein content and enzyme activity第30页
        2.3.2 Effects of Microwaves and Ultrasound on degree of hydrolysis第30-32页
        2.3.3 Effect on Enzymatic hydrolysis of GMP with different temperature, time, proteinconcentration and enzyme concentration第32页
        2.3.4 Effects of substrate concentration on DPP-V inhibitory activity第32-33页
        2.3.5 Effects of enzyme concentration, hydrolysis temperature and hydrolysis time on DPP-4inhibitory activity第33-34页
    2.4 Conclusion第34-36页
CHAPTER 3 Response surface optimization of dipeptidyal peptidase (DPP-IV) Inhibition ofglycomacropeptides hydrolysates第36-45页
    3.1 Introduction第36-37页
    3.2 Materials and methods第37-41页
        3.2.1 Materials第37页
        3.2.2 Experimental design of glycomacropeptide Concentrate Hydrolysate by CentralComposite (CCD)第37页
        3.2.3 RSM and Generation of the Optimum GMP Hydrolysate.第37-40页
        3.2.4 Enzyme hydrolysis and degree of hydrolysis第40页
        3.2.5 Determination of DPP-IV Inhibitory Activity第40-41页
        3.2.6 Statistical Analysis第41页
    3.3 Results and discussion第41-44页
        3.3.1 Hydrolysates Generated Within the Experimental Design第41-42页
        3.3.2 The Effects of different Factors affecting on the DPP-IV Inhibitory Activity of TheHydrolysate第42-43页
        3.3.3 Optimization and validation第43-44页
    3.4 Conclusion第44-45页
CHAPTER 4 Dipeptidyl peptidase-IV inhibitory peptides generated from Papain-Treated hydrolysis of aglycomacropeptide (cGMP20) protein第45-56页
    4.1 Introduction第45-46页
    4.2 Materials and methods第46-48页
        4.2.1 Materials第46页
        4.2.2 Preparation of the glycomacropeptide (GMP) hydrolysate第46页
        4.2.3 Separation and Purification of DPP-IV Inhibitory Peptides第46-47页
        4.2.4 Identification of DPP-IV Inhibitory Peptides by RP-HPLC-ESI-MS/MS第47页
        4.2.5 Assay of the DPP-IV-Inhibitory Activity第47-48页
        4.2.6 Peptide Synthesis第48页
        4.2.7 Peptide-cutter tool predicts enzymatic cleavage sequence第48页
    4.3 Results and discussion第48-54页
        4.3.1 DPP-4 inhibitory activity of GMP with different molecular weight ranges第48-49页
        4.3.2 Identification of peptide sequences第49-50页
        4.3.3 DPP-IV inhibitory activity of synthetic peptides第50-53页
        4.3.4 Peptide-cutter tool predicts enzymatic cleavage sequence第53-54页
    4.4 Conclusion第54-56页
CHAPTER 5 Effect of Glycomecropeptide (cGMP20) peptides on DPP-IV and GLP-1 in H716 cells第56-72页
    5.1 Introduction第56-57页
    5.2 Materials and methods第57-60页
        5.2.1 Materials and reagents第57页
        5.2.2 Instruments and equipments第57页
        5.2.3 Test design第57页
        5.2.4 cell culture第57-58页
        5.2.5 Deermination the number of cells第58页
        5.2.6 Cytotoxicity experiment第58页
        5.2.7 Treatment of different concentrations of glycomacropeptide peptides NCI-H716 cells第58-59页
        5.2.8 Extraction of DPP-4 and GLP-1 in cells第59页
        5.2.9 Determination of intracellular DPP-4 activity第59页
        5.2.10 Determination of GLP-1 secretion第59-60页
        5.2.11 Statistical analysis第60页
    5.3 Results and discussion第60-70页
        5.3.1 DPP-4 half inhibition rate of linagliptin and active peptide第60-61页
        5.3.2 Cell proliferation assay to determine optimal culture concentration第61页
        5.3.3 Cytotoxicity test to determine the optimum culture concentration of the sample to betested第61-63页
        5.3.4 Effect of sample to be tested on intracellular DPP-IV secretion第63-65页
        5.3.5 Effect of sample to be tested on intracellular GLP-1 secretion第65-70页
    5.4 Conculusion第70-72页
CHAPTER 6 Overall conclusions第72-74页
    6.1 Innovation points第73页
    6.2 Outlook第73-74页
References第74-80页
致谢 (Acknowledgement)第80-81页
作者简历 (Resume)第81-82页

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