ABSTRACT
In this study, Micrococcus species have been implicated as microbial population of fermented African oil bean seed ‘ugba’. The organism was catalase positive and could not utilize citrate and was coagulase and methyl red negative. Micrococcus isolates were screened for amylase enzyme production using the activity zone technique with iodine solution. Micrococcus sp was able to produce enzyme amylase and therefore could utilize the carbohydrate content of ugba. The quantity of amylase produced were relatively very small (0.824 IU/ml, 1.412 IU/ml, 1.427 IU/ml) which points that African oil bean seed contains relatively small amounts of carbohydrate. However, the enzyme producing ability of Micrococcus confirms that it plays a role in ugba fermentation and could contribute to its taste and flavor. Proper handling of ugba or a modern attempt in production may impend the participation of Micrococcus species in ugba spoilage and enzyme activity.
TABLE
OF CONTENTS
Title
page i
Certification ii
Dedication
iii
Acknowledgements iv
Table
of contents v
List
of tables viii
List
of figures ix
Abstract
x
Chapter one
1.0 Introduction 1
1.1 Aims and Objectives 3
Chapter two
2.0
Literature Review 4
2.1
Fermented Foods 4
2.2
Pentaclethramacrophylla 5
2.3
Uses and Application of Pentaclethramacrophylla 6
2.4
Methods of Production 8
2.4.1
Traditional Method 8
2.4.2 Laboratory Method 9
2.5
Microorganisms Involved in Ugba Fermentation 9
2.6
Micrococcus Spp 11
2.7 Description and Significance
of Micrococcus 12
2.8 Genome Structure of Micrococcus 12
2.9 Cell Structure and Metabolism
of Micrococcus 13
2.10
Ecology of Micrococcus 15
2.11 Pathology of Micrococcus 15
2.12 Pathogenesis of Micrococcus 16
2.13 Industrial Uses of Micrococcus 16
2.14
Amylase 17
2.15
Industrial Applications and Uses of Amylase 17
2.16
Microorganisms Involved in Amylase Production 18
Chapter Three
3.0 Materials and
Methods 19
3.1
Study Area 19
3.2
Sample Collection 19
3.3
Production of Ugba 19
3.4
Preparation and Sterilization of Media 20
3.5
Isolation of Micrococcus spp from
Ugba 20
3.6
Identification of Isolate 20
3.7
Gram Staining 21
3.8
Biochemical Tests 21
3.9
Screening of Micrococcus spp for
Amylase Production 23
3.10
Assay of Amylase Enzyme. 24
Chapter Four
4.0 Results 25
4.1
Identification of The Isolate 27
4.2
Growth and Amylase Production 27
Chapter Five
5.1
Discussion 28
5.2
Conclusion 29
References
LIST OF TABLES
Table Title Page
1 Medicinal
uses of Pentaclethramacrophylla 7
2.1
Characteristics of the Micrococcus spisolate. 25
2.2 Amylase
activities of Micrococcus spisolated
from ugba. 26
LIST OF FIGURES
Figure Title Page
1 African Oil
Bean Seed 6
2 Processes
Involved in Traditional Ugba Production 8
3 Tetrad arrangement of Micrococcus 14
4 Pigmented colonies of Micrococcus.
14
CHAPTER ONE
1.0
INTRODUCTION
Fermented foods whether
from plants or animal origin, are an intricate part of the diet of people in
all parts of the world. Fermented products remain of interest since they do not
require refrigeration during distribution and storage. The traditional
condiments have not attained commercial status due to the very short life,
objectionable packaging materials, stickiness and the characteristic putrid
odor (Oyedeji and Ijigbade, 2016).
Nigerian
indigenous fermented foods constitute a group of foods that are produced in
homes, villages and small-scale cottage industries. The fermented foods are
derived from substrates like roots, legumes, cereals, oil seeds, nuts, meat,
fish, milk, palm tree, sap, etc. (Iwuoha and Eke1996).These various plant seeds
that are fermented and used as food in some rural parts of Nigeria, are: ‘iru’
from African locust bean (Parkiabiglobosa),
‘ogiri’ from castor bean (Ricunuscommunis),
‘okpei’ from mesquite seed (ProsopsisAfricana)
and ‘ugba’ from African oil bean (Pentaclethramacrophylla)
(Enujiugha, 2009).
Ugba(orUkpaka
as it is called in Igbo language) popularly called African salad, is a
ready-to-eat food which is produced by the fermentation of African oil bean
seeds (Pentaclethramacrophylla).
African oil bean seed is from a woody plant predominant in the rain forest
areas of West and Central Africa belonging to the family Leguminosaea,
sub-family Mimosoidae (Anyanwu et al.,
2016).As an important nutritional item, Ugba is very rich in protein. The
production of ugba is usually produced as a family business that has become an
art that is handed over from one generation to another (Olasupoet al., 2016). It is an essential food
item for various traditional ceremonies where it is mixed with slices of boiled
stock fish (Ugba and Okpoloko). The natural fermentation of the seed which at
present is still done at the household level, renders the production
nutritious, palatable and non-toxic (Oyediji and Ijigbade, 2016).
Microbial
population of Ugba is introduced through the air, water, utensils, banana
leaves or the handler. No starter culture is used for the traditional method.
Microorganisms involved are predominantlyBacillus,
Micrococcus and Lactobacillus. Other organisms isolated from Ugba includes Pseudomonas, Staphylococcus, Enterobacters, Lueconostoc, CorynebactriumandAlkalegenes(Nwaguet al., 2011).These organisms utilize some of the nutrients
inherent in the seed while their metabolic activities detoxify the seed, soften
it and also produce flavor and aroma compounds which impact the characteristic
tastes and flavor (Nwagu and Amadi, 2013).
Micrococcus spdo
not play active role during microbial fermentation of Ugba (Nwaguet al., 2011). Bacillus and Micrococcus spp
(now Kocuriarosea) were especially
found to persist during the spoilage of Ugba, highly alkaline and unfavourable
conditions notwithstanding. And was able to utilize soluble starch and Bambara
flour for their growth and metabolic activities. The secretion of amylase (A
primary metabolite) was possibly induced in the presence of starch sources
which had to be hydrolyzed to simple sugar prior to their assimilation into the
cell (Nwagu and Amadi, 2013).
Carbohydrates
are the major nutrient content in Ugba and the ability to produce hydrolytic
enzymes signifies that Micrococcus spp
can break down these complex polymers to simpler units in order to assess
nutrients for their growth and metabolism (Nwagu and Amadi,2013).
Amylase
is a hydrolyzing enzyme in function which causes hydrolyses of molecules. In
biotechnology, amylases are the most important enzymes used. The main use of
amylase includes hydrolysis of starch to yield glucose syrup, amylase rich
flour and in the formation of dextrin during backing in food industries (Vermaet al., 2011). Amylase from microbial
sources can be produced in large quantities to meet the high industrial and
market demands, and the diversity of microbes as the source for bio-based
products is gaining ground every day (Adeoyoet
al., 2016).The interest in microbial amylase production is increasing
because of its economical bulk potentials in a wide range of industrial
applications and additives in food processing.
1.1 AIMS AND OBJECTIVES
i.
To isolate Micrococcus spfrom fermented ugba.
ii.
To assess the growth and
amylase production of Micrococcus spfrom
fermented Ugba.
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