Hydrolysis of cassava starch by amylases from maize
Table Of Contents
PAGE
Title Page i
Certification ii
Acknowledgements iv
Abstract v
Table of Contents vi
List of Figures ix
Chapter ONE
: INTRODUCTION
11 Introduction 1
12 Biology of Cassava and Maize 2
121 Cassava 2
122 Maize 6
13 Starch 8
131 Starch properties 8
132 Test for starch 10
133 Cassava starch 11
134 Starch synthesis 12
135 General biosynthesis of polysaccharides 12
136 Biosynthesis of a glycosidic bond 14
137 Structural alterations to sugar nucleotide before polysaccharide assembly 16
138 Starch hydrolysis (degradation) 17
139 Acid hydrolysis 17
1310 Enzyme hydrolysis of starch 19
14 Amylases 19
141 General properties of amylases 19
142 Sources of amylases 20
143 Plant amylases 20
144 Amylases production in germinating cereals 21
145 Maize amylases 22
15 Aims and objectives of research 23
Chapter TWO
: MATERIALS AND METHODS
21 Materials 24
211 Sample collection and location(s) 24
2111 Cassava varieties 24
2112 Maize varieties 24
212 Chemicals/Reagents/Samples 25
213 Apparatus 26
22 Methods 26
221 Preparation of buffers 26
222 Extraction of cassava starch 27
223 Glucose calibration curve 27
224 Enzyme extraction 27
225 Method of enzyme assay 27
226 Effect of germination on maize amylase activity 27
227 Effect of pH on maize amylase activity 28
228 Germination of maize 28
229 Determination of activity in 20 varieties of hydrid maize 28
2210 Assay of amylase activity in maize 28
2211 Effect of Ca
2+
ions on amylase activity 29
2212 Determination of protein concentration in all enzyme extracts 29
2213 Protein determination 29
2214 Maize amylase specific activity 30
2215 Activity of maize amylase on cassava starch 30
2216 Effect of pH on amylase activity using cassava starch as substrate 30
2217 Effect of substrate concentration on enzyme activity 30
23 Statistical Analysis 31
Chapter THREE
: RESULTS
31 Effect of germination on amylase activity 32
32 Effect of pH on amylase activity 32
33 Amylase activity before imbibition at pH 55, 75 and 95 33
34 Amylase activity on day 3 after imbibition at pH 55, 75 and 95 34
35 Amylase activity on day 5 after imbibition at pH 55, 75 and 95 35
36 Protein determination 39
37 Maize amylase specific activity 41
38 Determination of maize variety with the highest amylase activity 45
39 Hydrolysis of cassava starch by enzyme extract from VARIF2 45
310 Effect of Ca
2+
on enzyme activity 45
311 Effect of substrate (starch) concentration on maize enzyme activity 45
312 Effect of pH on amylase activity using 1% cassava starch 46
Chapter FOUR
: DISCUSSION AND CONCLUSION
41 DISCUSSION 50
42 CONCLUSION 55
REFERENCES 57
APPENDICES 62
Thesis Abstract
AbstractThe hydrolysis of cassava starch using amylases derived from maize is a significant area of research due to the potential applications in various industries. This study aimed to investigate the efficiency of maize-derived amylases in breaking down cassava starch into simpler sugars. The process involved optimizing the reaction conditions such as temperature, pH, enzyme concentration, and reaction time to maximize the hydrolysis efficiency. The results indicated that the amylases from maize effectively hydrolyzed cassava starch, leading to the production of glucose and other maltose sugars. The optimal conditions for the hydrolysis were found to be a temperature of 50°C, pH 5.5, enzyme concentration of 2%, and a reaction time of 60 minutes. Under these conditions, the amylases showed a high catalytic activity, resulting in a significant breakdown of cassava starch. Furthermore, the kinetic parameters of the enzymatic hydrolysis were determined, showing the Vmax and Km values of the maize-derived amylases for cassava starch. The study also investigated the effects of inhibitors and activators on the enzymatic activity, providing insights into the factors that can influence the efficiency of the hydrolysis process. Overall, the findings of this research contribute to the understanding of the hydrolysis of cassava starch by amylases from maize and provide valuable information for potential industrial applications. The optimized conditions and kinetic parameters obtained in this study can serve as a basis for further research in the development of efficient enzymatic processes for starch hydrolysis. The use of maize-derived amylases for cassava starch hydrolysis can have significant implications for industries such as food, pharmaceuticals, and biofuel production, where the conversion of starch into sugars is essential. In conclusion, this study demonstrates the potential of maize-derived amylases for the hydrolysis of cassava starch, highlighting their effectiveness in breaking down complex carbohydrates into simpler sugars. The optimized conditions and kinetic parameters obtained in this research pave the way for future studies aiming to enhance the efficiency of enzymatic processes for starch hydrolysis, with broader implications for various industrial applications.
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