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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

Abstract
The hydrolysis of cassava starch by amylases from maize is a significant area of research due to the applicability of these enzymes in various industries such as food processing, biofuel production, and pharmaceuticals. This study aimed to investigate the efficiency of amylases derived from maize in catalyzing the hydrolysis of cassava starch. The research involved the extraction and purification of amylase enzymes from maize using standard biochemical techniques. The enzymatic hydrolysis of cassava starch was carried out under controlled conditions of pH, temperature, and enzyme concentration. The progress of starch hydrolysis was monitored by measuring reducing sugar content using the dinitrosalicylic acid (DNS) method. The effects of varying enzyme concentrations, pH levels, and temperatures on the hydrolysis rate were evaluated to determine the optimal conditions for the enzymatic reaction. The kinetic parameters of the enzymatic hydrolysis process were determined by fitting the experimental data to Michaelis-Menten kinetics and Lineweaver-Burk plots. The results indicated that the maize-derived amylases effectively hydrolyzed cassava starch, with higher enzyme concentrations leading to increased reaction rates. Optimal pH and temperature ranges for the enzymatic hydrolysis were identified, showing that the enzymes exhibited the highest activity under specific conditions. The kinetic analysis revealed the maximum reaction rate and Michaelis constant for the enzymatic hydrolysis, providing insights into the catalytic efficiency of the maize-derived amylases on cassava starch. Furthermore, the stability of the amylase enzymes under different pH and temperature conditions was assessed to understand their robustness and potential industrial applications. The findings of this study contribute to the understanding of utilizing maize-derived amylases for efficient hydrolysis of cassava starch, offering insights into improving enzymatic processes in various industrial sectors. Overall, this research provides valuable information on the enzymatic hydrolysis of cassava starch by amylases from maize, highlighting the importance of enzyme optimization for enhancing process efficiency and sustainability in biotechnological applications.

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