پایان نامه تولید آنزیم پروتئاز در بیو راکتور سینی‌ دار با استفاده از فرآیند تخمیر حالت جامد

فهرست:

فصل1  1

مقدمه  1

1-1. مقدمه. 2

1-2. تعریف آنزیم.. 2

1-3. تاریخچه آنزیم.. 2

1-4. ساختار آنزیم.. 4

1-5. تقسیم‌بندی آنزیم‌ها 5

1-6. تاریخچه آنزیم پروتئاز 6

1-7. عملکرد پروتئازها 6

1-8. تخمیرحالت جامد. 7

1-9. ضرورت انجام پروژه. 8

1-10. اهداف این پروژه. 8

فصل2 مروری بر منابع مطالعاتی   10

2-1. مقدمه. 11

2-2. پروتئازها 11

2-3. منابع پروتئازها 12

2-3-1. پروتئازهای گیاهی.. 12

2-3-2. پروتئازهای حیوانی.. 13

2-3-3. پروتئازهای میکروبی.. 13

2-4. تقسیم بندی پروتئازها 16

2-5. پروتئازهای قلیایی.. 19

2-6. مکانیزم عمل پروتئازها 22

2-7. کاربردهای صنعتی آنزیم پروتئاز 22

2-7-1. صنعت مواد شوینده. 23

2-7-2. صنایع غذایی.. 24

2-7-3. صنعت چرم. 25

2-7-4. صنعت عکاسی.. 26

2-7-5. صنایع دارویی.. 26

2-7-6. مدیریت محیط زیست... 27

2-8. تولید آنزیم پروتئاز 27

2-9. تخمیر حالت غوطه ور 28

2-9-1. تخمیر حالت جامد. 29

2-9-2. مقایسه سیستم‌های تخمیر جامد و غوطه‌ور 29

2-9-3. انتقال جرم در تخمیر حالت جامد. 30

2-9-4. عملیات انتقال جرم در مقیاس ماکرو. 31

2-9-5. عملیات انتقال جرم در مقیاس میکرو. 32

2-9-6. انتقال اکسیژن.. 32

2-9-7. نفوذ آنزیم‌ها 33

2-9-8. جنبه‌های انتقال حرارت... 34

2-9-9. میکروارگانیزم‌های مورد استفاده در تخمیر حالت جامد. 35

2-9-10. کاربردهای تخمیر حالت جامد. 37

2-9-11. آنزیم‌های بدست آمده از فرآیند تخمیر جامد. 38

2-10. طراحی بیوراکتور 39

2-11. انواع بیوراکتورهای مورد استفاده در تخمیر حالت جامد. 40

2-11-1. بیوراکتورهای سینی‌دار 41

2-11-2. بیوراکتورهای بستر‌پرشده 42

2-11-3. بیوراکتورهای استوانه ای‌دوار 43

2-11-4. بیوراکتورهای بستر‌سیال.. 45

2-12. مراحل عمومی برای انجام فرآیند SSF در داخل بیوراکتور 46

2-13. عوامل مؤثر در تولید پروتئاز در فرآیند SSF در داخل بیوارکتور 47

فصل3 مواد و روش‌ها 48

3-1. مقدمه. 49

3-2. تجهیزات مورد استفاده. 49

3-3. تعیین مشخصات سوبسترا 50

3-3-1. محاسبه میزان خاکستر. 50

3-3-2. محاسبه میزان رطوبت... 51

3-3-3. محاسبه میزان قند موجود در سوبسترا 51

3-3-4. محاسبه میزان پروتئین.. 52

3-3-5. تعیین درصد مواد استخراجی.. 54

3-3-6. تعیین درصد سلولز. 54

3-3-7. تعیین درصد لیگنین.. 55

3-3-8. تعیین درصد همی‌سلولز. 55

3-3-9. محاسبه اندازه ذرات سوبسترا 55

3-4. میکروارگانیسم و محیط کشت... 56

3-4-1. انتخاب میکروارگانیسم.. 56

3-4-2. مشخصات میکروارگانیسم.. 57

3-4-3. محیط کشت... 57

3-4-4. تهیه مایه تلقیح.. 59

3-4-5. منحنی رشد باکتری... 60

3-4-6. تعیین pH بهینه باکتری... 60

3-5. تخمیر حالت جامد. 61

3-6. نمونه‌گیری و استخراج آنزیم از سوبسترای تخمیر‌یافته. 63

3-7. فعالیت پروتئاز 64

3-7-1. منحنی استاندارد تیروزین.. 65

3-8. بررسی تأثیر پارامترهای مختلف بر روی تولید آنزیم پروتئاز در بیوراکتور سینی‌دار 66

3-8-1. تأثیر نوع سوبسترای جامد. 66

3-8-2. تأثیر مدت زمان تخمیر. 67

3-8-3. اثر دما 67

3-8-4. تأثیر pH... 67

3-8-5. اثر پارامترهای مختلف بر روی استخراج آنزیم.. 68

3-8-6. تأثیر رطوبت اولیه سوبسترا 68

3-8-7. تأثیر رطوبت داخلی راکتور 68

3-8-8. تأثیر اندازه ذرات... 68

3-8-9. تأثیر میزان تلقیح.. 69

3-8-10. تأثیر غنی‌سازی سوبسترا با منابع کربنی و نیتروژنی.. 69

3-8-11. تأثیر pH بر فعالیت و پایداری آنزیم تولیدی... 69

3-8-12. تأثیر دما بر فعالیت و پایداری آنزیم تولیدی... 70

3-9. کاربردهای آنزیم تولیدی... 71

3-9-1. افزودنی به مواد شوینده. 71

3-9-2. پردازش چرم. 71

3-9-3. هیدرولیز لایه ژلاتینی فیلم‌های عکاسی و آزاد سازی نقره. 72

3-10. مقایسه تولید آنزیم پروتئاز در بیوراکتور و فلاسک.... 72

فصل4 نتایج و تفسیر آنها 73

4-1. مقدمه. 74

4-2. محاسبه خصوصیات سبوس گندم. 74

4-3. منحنی رشد باکتری... 75

4-4. pH بهینه رشد باکتری... 75

4-5. بررسی پارامترهای مختلف بر تولید پروتئاز 76

4-5-1. تأثیر مدت زمان تخمیر. 76

4-5-2. بررسی تأثیر نوع سوبسترای جامد. 78

4-5-3. بررسی پارامترهای مؤثر بر استخراج پروتئاز 79

4-5-4. تأثیر pH ابتدایی.. 82

4-5-5. بررسی دمای داخل بیوراکتور 82

4-5-6. تأثیر رطوبت ابتدایی سوبسترا 84

4-5-7. تأثیر رطوبت داخلی بیوراکتور 85

4-5-8. تأثیر اندازه ذرات... 86

4-5-9. تاثیر میزان مایه تلقیح.. 87

4-5-10. بررسی تأثیر غنی سازی سوبسترا با منابع کربنی و نیتروژنی.. 87

4-6. بهینه سازی شرایط فعالیت پروتئازی آنزیم.. 92

4-6-1. تعیین pH بهینه فعالیت آنزیم.. 92

4-6-2. تعیین دمای بهینه فعالیت آنزیم.. 94

4-6-3. تعیین pH پایداری آنزیم.. 95

4-6-4. تعیین دمای بهینه پایداری آنزیم.. 96

4-7. کاربردهای آنزیم پروتئاز قلیایی حاصل از B.licheniformis. 97

4-7-1. عملکرد پروتئاز قلیایی به عنوان افزدونی به شوینده. 97

4-7-2. موزدایی از پوست... 98

4-7-3. هیدرولیز لایه ژلاتینی فیلم‌های X-Ray. 99

4-8. مقایسه تولید آنزیم پروتئاز در بیوراکتور و فلاسک.... 100

فصل5 نتیجه‌گیری و پیشنهادها 103

5-1. نتیجه‌گیری... 104

5-2. پیشنهادها 106

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