ABSTRACT
The Isolation and characterization of amylolytic bacterial strains with hydrolytic profile during anaerobic digestion of the mixture of cow rumen, dried cassava peels and poultry droppings was the subject of this research. Hydrolysis is important to biogas production as it involves breakdown of polysaccharides present in substrates. The experiment lasted for 30 days using a 10 litre scale bio-digester. Hydrolytic amylase producing bacteria isolated were Micrococcus luteus, Pseudomonas aeruginosa, Klebsiella oxytoca, Aspergillus spp, and Bacillus subtilis. The total anaerobic plate count ranged from 2.57cfu/g to 3.53cfu/g and the observed pH ranged from 5.12 to 6.19. The study reveals that the mixture of cassava peels, poultry droppings and cow rumen has the potential of producing biogas. In conclusion, further investigation of hydrolytic groups of bacteria beyond 30 days of anaerobic retention period should be investigated.
TABLE OF CONTENTS
Title Page i
Certification ii
Dedication iii
Acknowledgement iv
Table of Contents v
List of Tables ix
Abstract x
Chapter One
1.0
Introduction 1
1.1 Aim
and Objectives 2
Chapter Two
2.0 Literature Review 3
2.1 What is Biogas? 4
2.2 Biogas Plants 5
2.3 Typical Composition of Biogas 6
2.4 Use and Application of Biogas 6
2.5 Microorganisms Involved in Biogas
Production 6
2.6 Amylolytic Bacteria 7
2.7 Biomass and Biogas Formation 8
2.8 Factors
Affecting the Hydrolytic Activity In Biogas Plants: A Brief Overview 10
2.8.1 Temperature 10
2.8.2 pH 10
2.8.3 Substrate Related Factors 11
2.8.3.1 Particle Size 11
2.8.3.2 Substrate
Accessibility and Substrate Structure 12
2.9 Decomposition of Organic Compounds in the
Biogas Process 12
2.9.1 Stage 1. Hydrolysis 13
2.9.1.1 Hydrolysis
of Polysaccharides 14
2.9.1.2
Hydrolysis of Proteins 15
2.9.1.3
Hydrolysis of Fats 16
2.9.2 Stage 2. Fermentation 16
2.9.3 Stage 3. Anaerobic Oxidation 19
2.9.4 Stage 4. Methane Formation 21
2.10 Alternative Methane Production Pathway from
Acetate 23
2.11 Diversity of Microorganisms 24
2.12 Interactions between the Various Microbial
Groups 25
Chapter Three
3.0 Materials
and Methods 26
3.1 Sample
Collection 26
3.2 Isolation
and Assessment of Bacterial Populations 26
3.3 Characterization
and Identification of the Isolates 26
3.3.1 Identification of Bacteria 26
3.3.2 Biochemical Tests 27
3.3.2.1 Catalase Test 27
3.3.2.2 Coagulase Test 27
3.3.2.3 Oxidase Test 27
3.3.2.4 Citrate Utilization Test 28
3.4 Bio-Digester
Design And Loading 28
Chapter Four
4.0. Results 30
Chapter Five
5.1. Discussion 33
5.2. Conclusion 34
5.3. Recommendation 34
References 35
LIST
OF TABLES
S/N TABLE PAGE NO
4.1
Total Anaerobic count (TAC) of Fresh cassava peels, cow rumen and poultry
droppings 31
4.2 physicochemical parameter of fresh
cassava peels, cow rumen and poultry droppings 32
4.3 Characteristics feature of isolated
bacteria 33
4.4 Biochemical Characteristics of the
isolated biogas production 34
CHAPTER ONE
1.0 INTRODUCTION
Biogas production technology through
anaerobic digestion is an alternative form of fuel and a good fertilizer
source. Biogas production is achieved via anaerobic digestion. The digestion
process encompasses four stages namely: hydrolysis, acidogenesis, acetogenesis
and methanogenesis Zieminski et al. ( 2012).
Cellulolytic” and “Amylolytic” are two
common terms referring to anything capable of breaking down cellulose and
amylose respectively. Enzymes responsible for the breakdown of cellulose and
amylose are called “cellulase” and “amylase” respectively. Common cellulolytic
and amylolytic bacteria in the hydrolytic stage of biogas production include
the genera Bacteriodes, Clostridium and Acetivibrio (Cirne et al., 2007; Doi,
2008).
Amylolytic
enzymes are widely distributed among microbes. They have been extracted from
several fungi, yeasts and bacteria, however, enzymes from Fungal and bacterial
sources have dominated applications in industrial Sectors. Bacillus and Aspergillus
spp. are known as the most active amylase Producers. With the advent of new
Boundaries in biotechnology, the amylase Family of enzymes and particularly
those From microbial sources, have found Potential application in a number of
Various fields, such as analysis (glycogen Determination), industries
(including Brewing, baking, starch processing, Detergent, textiles, paper,
food) and Clinical or pharmaceutical applications (diagnostic aids) (Pandey et
al., 2000; Gupta et al., 2003). The major advantages of using Microorganisms
for the production of Amylases are their economical bulk Production capacity
and stability with respect to pH and temperature. Furthermore, microbes are
easy to Manipulate in regard to obtaining enzymes with the desired
characteristics. Amylolytic enzymes play an important Role in the degradation
of starch, and are Produced in bulk from microorganisms, Representing about 25-33%
of the world Enzyme market (Ramesh and Lonsane, 1990; Konsula and Liakopoulou-
Kyriakides, 2004; Sivaramakrishnanet al., 2006). In the case of energy
costs, Worldwide interest has been growing in Recent years regarding the
acquisition of Desirable amylases that are capable of Digesting raw starches
during the process Of starch liquefaction, but without the need For heating,
inevitably leading to the Effective utilization of biomass and Reduction in the
total cost of starch Processing (Wang et al., 1999; Robertson Et al.,
2006). The microbial amylases that are used for Industrial purposes are
derived mainly From Bacillus licheniformis, Bacillus
Amyloliquefaciens and Aspergillus oryzae. Accordingly, native
amylolytic microbial Strains especially, have abundant Applications to relevant
industries in Biotechnology (Nigam and Singh. 1995; Crabb and Mitchinson.
1997).
1.1 AIM AND OBJECTIVES
To screen and isolate amylolytic bacteria involved
in biogas production while the specific objectives are:
·
To
isolate bacteria from wastes (cow rumen, cassava peels and poultry droppings)
·
To
screen the isolate for amylase activity
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