發(fā)布時(shí)間：2020-12-22 13:37

　　蕓豆（Phaseolus vulgaris）淀粉酶抑制劑亞型1（α-AI1）可被廣泛用來(lái)輔助治療肥胖癥與糖尿病。通過(guò)淀粉酶抑制劑合適的結(jié)構(gòu)、物理化學(xué)性質(zhì)、功能性可影響腔內(nèi)邊緣淀粉酶的活力。最終結(jié)果是降低餐后血高血糖與胰島素水平,增加淀粉消化的抗性和腸內(nèi)微生物的活力。然而,淀粉酶抑制劑提取物的療效及安全性取決于加工及提取工藝。抑制劑可被添加到食品中,用來(lái)增加糖尿病人的碳水化合物耐受量,降低攝取的能量,從而避免出現(xiàn)肥胖癥和提高抗性淀粉的含量。一般的粗提工藝包括水提取,pH分離和乙醇（75%）沉淀,它可用來(lái)篩選抑制劑提取合適的品種。幾種蕓豆中,紅蕓豆中淀粉酶抑制劑的活力最高,黑蕓豆的則最小。隨后的粗提取物的SDS-PAGE電泳圖顯示蛋白質(zhì)組分的數(shù)量減少了,只有兩條主要帶。1.4-2.6克紅蕓豆粗提物的特異性血凝活性平均為14222.2U/mg。進(jìn)一步采用DEAE-Sepharose CL-6B和Sephacryl S-200層析柱分離純化。在0.09%原蕓豆粉（w/w）的基礎(chǔ)上通過(guò)電泳純化得到的淀粉酶抑制劑檢測(cè)不到血凝活性,同時(shí),比酶活由0.32增加到80.78 U/mg。Native-PA... 

【文章來(lái)源】：江南大學(xué)江蘇省 211工程院校 教育部直屬院校

【文章頁(yè)數(shù)】：82 頁(yè)

【學(xué)位級(jí)別】：碩士

【文章目錄】：
ACKNOWLEDGEMENTS
DEDICATION
ABSTRACT
摘要
TABLE OF CONTENTS
ABBREVIATIONS USED
CHAPTER 1:INTRODUCTION & LITERATURE REVIEW
    1.1 Introduction:Kidney Bean Starch Blockers
    1.2 Literature Review
        1.2.1 Favorable Physico-chemical Properties of the Phaseolus vulgaris α-amylase inhibitor
            1.2.1.1 Structural properties of the Phaseolus vulgaris α-amylase Inhibitor
            1.2.1.2 Factors that affect the Phaseolus vulgaris α-amylase inhibitor Activity
        1.2.2 The Starch Blocking Mechanism of the Phaseolus vulgaris Alpha-amylase Inhibitor
        1.2.3 Efficiency of α-AI1 Extracts in Reducing Activity of Amylases in Man
        1.2.4 The Beneficial Effects of the Phaseolus vulgaris α-amylase Inhibitor
            1.2.4.1 Decreased obesity due to Phaseolus vulgaris α-amylase extracts
            1.2.4.2 The anorexigenic effect of Phaseolus vulgaris α-AI1 extracts
            1.2.4.3 Reduced postprandial plasma hyperglycemia and insulin due to α-AI1 extracts
        1.2.5 Safety and Toxicity of the Phaseolus vulgaris α-amylase Inhibitor Extracts
            1.2.5.1 Toxic effects Associated with kidney beans（Phaseolus vulgaris）
            1.2.5.2 Acute toxicity studies
            1.2.5.3 Chronic toxicity studies
            1.2.5.4 Prospective research on beneficial effects:the potential of α-amylase inhibitor isoform 1 extracts against colorectal cancer
    1.3 Justification and Objectives of Study
        1.3.1 Justification
        1.3.2 Objectives of study
        1.3.3 Hypothesis of Study
CHAPTER 2:VARIETY SELECTION,PURIFICATION AND PARTIAL CHARACTERISATION OF PHASEOLUS VULGARIS ALPHA-AMYLASE INHIBITOR（α-AI1）
    2.1 Introduction
    2.2 Materials
    2.3 Methods
        2.3.1 Determination of Protein Content
        2.3.2 Stabilizing α-amylase activity of Porcine Pancreatic α-amylase
        2.3.3 Porcine Pancreatic α-amylase and Inhibitor Activity Measurement
        2.3.4 Crude Extraction of the Phaseolus vulgaris α-amylase inhibitor（α-AI1） and Variety Selection
        2.3.5 Chromatographic Purification Phaseolus vulgaris α-amylase inhibitor（α-AI1） from Speckled Kidney BeanVariety
        2.3.6 Purity Assessment
            2.3.6.1 SDS page analysis
            2.3.6.2 Native page of purified inhibitor
            2.3.6.3 Hemagglutinating activity
        2.3.7 Partial Characterization of Purified Speckled Kidney bean（Phaseolus vulgaris） α-amylase Inhibitor（α-AI1）
        2.3.8 Statistical Analysis
    2.4 Results and Discussion
        2.4.1 Stabilizing of Porcine Pancreatic α-amylase Catalytic Activity
        2.4.2 Variety Selection through Crude/Partial Extraction from Different Varieties
        2.4.3 Chromatographic Purification of Phaseolus vulgaris α-amylase inhibitor（α-AI1） from Speckled kidney Beans
        2.4.4 Partial Characterization of the Purified Inhibitor from Speckled Kidney Beans
            2.4.4.1 Effect of inhibitor concentration on activity of speckled kidney bean（Phaseolus vulgaris） α-amylase inhibitor（α-AI1） activity:Determination of IC50 value
            2.4.4.2 Effect of pH on the activity of the Phaseolus vulgaris α-amylase inhibitor from speckled kidney beans
            2.4.4.3 Effect of Incubation time on the activity of the Phaseolus vulgaris α-amylase inhibitor from speckled kidney beans
    2.5 Conclusions
CHAPTER 3:HEAT STABILITY OF THE PHASEOLUS VULGARIS ALPHA AMYLASE（α-AI1） INHIBITOR FROM RED SPECKLED KIDNEY BEANS
    3.1 Introduction
    3.2 Materials
    3.3 Methods
        3.3.1 Purification the Phaseolus vulgaris α-amylase Inhibitor（αAI1）
        3.3.2 Purity Assessment
            3.3.2.1 SDS-PAGE analysis
            3.3.2.2 Native-PAGE of purified inhibitor
            3.3.2.3 Hemagglutinating activity
        3.3.3 Determination of Protein Content
        3.3.4 PPA and Inhibitor Activity Assessment
        3.3.5 DSC Calorimetry Measurements
        3.3.6 Heat Stability Treatments
            3.3.6.1 Treatments for Response Surface Methodology experiments
            3.3.6.2 Heat treatment for effects of hofmeister series salts and polyols intrinsic fluorescence and ultraviolet absorption spectra
        3.3.7 Intrinsic Fluorescence and Ultraviolet Absorption Spectra
        3.3.8 Surface Hydrophobicity Measurements
        3.3.9 Experimental Design and Statistical Analysis
            3.3.9.1 Response Surface methodology design
            3.3.9.2 Statistical analysis
    3.4 Results and Discussions
        3.4.1 Model Selection and Residual α-amylase Inhibitory Activity Variation
        3.4.2 DSC Analysis:Phaseolus vulgaris α-amylase Inhibitor（α-AI1）
        3.4.3 Molecular Changes Involved in Activity Loss
        3.4.4 Effect of Selected Hofmeister Series Salts and Polyols
    3.5 Conclusion
CHAPTER 4:GENERAL CONCLUSIONS,RECOMMENDATIONS AND REFERENCES
    4.1 General Conclusions
    4.2 Recommendations
    4.3 Published Papers
    4.4 Appendix
    4.5 List of Figures and Tables
    4.6 References



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