Home / Biochemistry / Hydrolysis of cassava starch by amylases from maize

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
This research project focused on the hydrolysis of cassava starch using amylases extracted from maize. The study aimed to investigate the efficiency of maize amylases in breaking down cassava starch into simpler sugars. The process of hydrolysis was conducted under controlled conditions to observe the impact of enzyme concentration, pH, and temperature on the rate of starch conversion. Various analytical methods were employed to monitor the progress of hydrolysis, including reducing sugar assays and starch content measurements. The results of the study indicated that maize amylases were effective in catalyzing the hydrolysis of cassava starch. Higher enzyme concentrations led to an increased rate of starch conversion, highlighting the importance of enzyme dosage in the hydrolysis process. The optimal pH for amylase activity was found to be slightly acidic, with variations outside this range resulting in reduced efficiency. Temperature was also a critical factor influencing the hydrolysis rate, with an increase in temperature generally accelerating the process up to a certain point. Beyond the optimal temperature, enzyme denaturation occurred, leading to a decrease in hydrolysis efficiency. The study identified the temperature range within which maize amylases exhibited the highest activity on cassava starch, providing valuable insights for industrial applications. Moreover, the analysis of the hydrolysis products revealed the presence of glucose and maltose, indicating the successful breakdown of cassava starch into simpler sugars. The quantification of reducing sugars confirmed the extent of starch conversion by maize amylases. These findings contribute to the understanding of enzymatic hydrolysis processes and offer practical implications for the bioconversion of starch into valuable products. Overall, this research demonstrates the potential of maize amylases as effective biocatalysts for the hydrolysis of cassava starch. The optimization of enzyme concentration, pH, and temperature parameters can enhance the efficiency of starch conversion, providing a sustainable approach for bio-based industries. Further studies could explore the application of maize amylases in large-scale starch hydrolysis processes and investigate their compatibility with other enzymatic systems for improved saccharification yields.

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