Download Microbial and enzymatic changes associated with the production of

Sky Journal of Food Science Vol. 2(2), pp. 10 - 18, February, 2013
Available online http://www.skyjournals.org/SJFS
©2013 Sky Journals
Full Length Research Paper
Microbial and enzymatic changes associated with the
production of Ogiri from castor oil bean using B.
subtilis as starter culture
Ojinnaka, M. C.1*, Ojimelukwe, P. C2 and Ezeama C. F2
1
Department of Food Science and Technology, Imo State University, Owerri,
P. M. B. 2000 Owerri, Nigeria.
2
Department of Food Science and Technology, Michael Okpara University of Agriculture, Umudike, P. M. B 7267
Umuahia, Abia State, Nigeria.
Accepted 6 February, 2013
Various types of microorganisms were isolated from traditionally fermented castor oil bean Ricinus communis.
Ten isolates were obtained from a 96 h spontaneously fermented castor oil bean to produce ogiri. Pure cultures
of the isolates were identified employing API 50 CHB/E, API 20 E and API 50 CHL media. Their identities were
confirmed as B. subtilis, B.circulans 2, B. stearothermophilus, Brevibacilus brevis, B. megaterium 2, L.
pentosus, and L. plantarum. However B. subtilis was found to be predominant and was used as a monoculture
starter for the production of three different fermented castor oil bean samples: B1( 0% NaCl/Lime), B2 (2% NaCl),
B3 (3% Lime). The three fermented castor oil bean samples were assayed using High Performance Liquid
Chromatography for enzymes. There was gradual increases in the activities of the proteinase enzyme though
there were no significant difference (P>0.05) in the fermented castor oil bean samples. All the three samples
showed increases in proteinase activity as fermentation period progressed. The proteinase values were 2.07 –
3.02 µg/ml for sample B1, 1.38 – 2.40 µg/ml for B2 and 1.74 – 2.09 µg/ml for B3. There were no significant
difference (P>0.05) in the lipase content in the three fermented castor oil bean samples. The lipase activity
values were B1: 5.69 – 7.01 µg/ml, B2 : 2.99 – 4.79 µg/ml and B3 :2.22 – 4.92 µg/ml. There was significant
difference (P<0.05) in the amylase activity during the fermentation of castor oil bean into ogiri. There was a
steady increase in the amylase activity of sample B1 (3.14 – 4.81 µg/ml)while those of B2 (1.90 – 3.01 µg/ml) and
B3 (2.23 – 2.87 µg/ml ) experienced fluctuations in their amylase activity but increased steadily from 72h
fermentation. But in all the three samples B1 had higher enzyme activity value than B2 and B3..
Key words: Ogiri, castor oil bean, fermentation, enzymes.
INTRODUCTION
Ogiri is a product of traditional alkaline fermentation of
castor oil bean consumed as food condiment in eastern
Nigeria. The castor oil plant Ricinus communis, also
known as Palma(e) christi or wonder tree, is a perennial
scrub of the spurge family Euphorbiaceae. R. communis
probably originates from Africa and was used in ancient
Egypt and by the Romans and Greeks (Serpico and
White, 2000; Weiss, 2000).
*Corresponding Author. E-mail: [email protected].
Castor seeds are a rich source of oil which can be
extracted by milling, boiling, pressing or solvent
extraction. Apart from medical applications, the oil has
long been used as an inexpensive fuel for oil lamps.
Because of its high proportion of the fatty acid ricinoleic
acid, today it is a valued industrial raw material for
lubricants, paints, coats, cosmetic products and many
more (Ogunniyi, 2006; Mutlu and Meier, 2010).
Interestingly in western Africa, alkaline-fermented castor
seeds are part of the flavoring soup condiment ogiri
(Parkouda et al., 2009). Ogiri which is mainly produced
by women constitute an economical source for the
producers. It is used as substitute for meat or fish for
Ojinnaka et al.
flavouring soups and can serve as a nutritious condiment
in the diet and its consumption should be encouraged.
Previous studies have shown that the main
microorganisms involved in the fermentation of castor oil
bean into ogiri are Bacillus especially Bacillus subtilis (Ibe
and Orabuike, 2009). However, this study seeks to
monitor the microbial and enzymatic activities in the
fermentation of ogiri from castor oil bean (R. communis)
using B. subtilis as a monoculture starter for 96 h.
11
gas pack (oxoid) at 35 ± 2°C and incubated for 24 - 48 h.
The plates were observed for bacteria growth after
incubation. Colonies observed were counted and
recorded. The bacteria colonies were randomly picked
and sub-cultured repeatedly unto fresh agar plates to
obtain pure culture respectively. The isolates were
identified or characterized based on their cultural
characteristics, cell morphology and biochemical
characteristics using API kits (Analytical Profile Index)
(Biomereux).
MATERIALS AND METHODS
Identification of organisms
The castor bean seeds (R. communis) used in this
research was purchased from New Aba market in Abia
State, Nigeria. All reagents and media used in the
research work were obtained from major commercial
suppliers and they were all of analytical grade.
Traditional preparation of fermented castor oil bean
seed (ogiri)
The fermented castor oil bean seeds (ogiri) used for
isolation of microorganisms was prepared
using the
method of Ibe and Orabuike (2009). About 1.5 kg of
castor bean seeds were boiled for two hours, dehulled,
drained and boiled again for about 6 h. The seeds were
ground in a clean mortar into paste, then wrapped with
aluminum foil and allowed to ferment for four days at
room temperature (29°C).
Identification of the isolates were carried out using the
API strips according to manufacturer’s instructions (API
50 CHB/E medium and API 20 E medium for Bacillus
species while API 50 CHL medium was used for
Lactobacillus species, - Montalieu, Vercieu France).
Preparation of inocula
fermentation of ‘ogiri’
used
in
laboratory
Nutrient broth (Oxoid CM3 Unipath, Basingstoke, UK)
were poured into test tubes and sterilized at 12°C for 15
min. They were allowed to cool to 25°C. With a sterile
wire loop B. subtilis from the agar slants was transferred
into 15ml of Nutrient broth in test tubes and incubated at
37°C for 24 h. The cell suspensions obtained were
shaken together and 2 ml was taken using a sterile
syringe for inoculation on the sterilized mashed castor
bean samples.
Determination of microorganisms associated with
fermented castor bean seed (ogiri)
Preparation of ‘ogiri’ from R. communis using starter
culture
The isolation of microorganism from traditionally
fermented ‘ogiri’ sample was stepwise. 1 g of ‘ogiri’ was
weighed on mettler balance and transferred into 9.0 ml
-1
sterile Ringer solution (10 dilution). The sample was
properly mixed by shaking with sterile 1.0 ml pipette. One
(1.0) ml was taken from dilution 10-1 and was diluted ten
fold serial dilution up to 10-10 dilutions.
Isolation and identification of the isolates
One milliliter (1.0ml) aliquots of appropriate dilutions were
plated out or inoculated into disposable petri dishes in
duplicates respectively, while molten cooled MRS agar
and plate count agar were poured onto the inoculum
using pour plate method. The plates were properly and
uniformly mixed. The plates were allowed to set and the
PCA plates were incubated at 35 ± 2°C for 24 h, while
MRS agar plates were incubated in an anaerobic jar with
The laboratory fermentation of castor bean was done
using the method of Enujiugha (2009).
Determination of enzymes in fermenting castor bean
using HPLC using waters 616/626 HPLC
The analysis was carried out using Waters 616/626
HPLC inter-phased with a computer system. The
samples were weighed into a set of homogenized tubes
and homogenized for 30 min. A quantity of 2.5 g of the
homogenized samples were transferred to a set of
extraction tubes. 10.0 ml ultrapure water was added,
followed by 10ml of acetone containing 5% (V/V) glycerol
and 1.5% 2-mercaptoethanol. The set up was shaken on
mechanical shaker (Edmund Buhler Model, USA). The
sample solutions were quickly transferred to a set of
polypropycene containers and caped properly. The
samples were again shaken for 15 minutes, transferred to
the centrifuge and centrifuged for 20 min at 500 rpm.
12
Sky J. Food Sci.
Then samples were put into a set of auto-analyser tubes
caped and stored for enzyme analysis using Waters
616/626 HPLC.
The accessories involved in the separation of the various
analytes of the enzymes are:
i.) Column (stationary phase): Is Lichrosorb Si-60 7 um.
ii.) The mobile phase was isooctane / tertbutyl methyl
ether (97.3ml).
iii.) The detector was fluorescence detector (330 nm).
iv.) Interphased with a Digital Converter Software (DCS).
v.) Flow rate used was 1.5 ml/minutes.
The sequence of separation was as follows:
The sample solutions were arranged in a set of
autoanalyser cups and mounted on the auto-sampler. All
the necessary data/information were entered on the
sample table of the softwares used (that is, the laboratory
numbers, the weight of sample, dilution factor, range of
standards, concentrations, and all the information were
saved accordingly.
The HPLC and the Autoanalyser sampler were put on.
The probe of the auto-sampler picked the sample solution
into the stationary phase(Is Lichrosorb Si-60 7 um). The
sample solution on suction to the stationary phase, mixed
with the mobile phase (Isooctane/tertbutyl methyl ether
(97:3)ml. The sample mixture with the mobile phase
moved along the stationary phase. The analytes were
eluted according to their molecular weight into a
Fluorescence detector (FD) at the wavelength of 230nm.
The intensity of the eluted analyte is fed on the Digital
Converter Software 1473 series (DCS) Waters, starting
from the analytes of a set of standards, and followed by
the samples analytes.
Calculation
The software stored the signal intensity from the standard
concentration at the standards (0.0, 0.20, 0.4, 0.6, 0.8
ug/ml), and uses it to calculate the concentration of the
unknown.
mg/ml enzyme (in Extract) = Dilution factor x peak height
response
mg/ml enzyme (in sample) = ug/m/in extract x volume
Wt of sample
Statistical analysis
Statistical analysis was done for each set of data
obtained following the procedures of Steel and Torie
(1984) for a Factorial Randomized Complete Block
Design (Factorial RCBD) while GENSTAT discovery
package (2006 edition) was used for the analysis of the
data. Comparison of treatment means and significant
differences between treatment means separated using
Fisher’s Least Significant Difference (LSD) as outlined by
Gomez and Gomez (1984).
RESULTS AND DISCUSSION
Tables 1, 2 and 3 show the identified organisms from
fermented castor oil bean. The fermentation of castor oil
bean seeds into ogiri, (a fermented food condiment of
eastern region of Nigeria) was found to be caused by
Bacillus species of which B. subtilis was predominant,
though some lactic acid bacteria were also isolated from
the ogiri condiment. Some species of Bacillus and Lactic
acid bacteria isolated from the traditionally fermented
castor oil bean mash using API kit software include B.
subtilis, B. circulans 2, B. stearothermophilus, B. brevis,
B. megaterium 2, L. pentosus, L. plantarum and L.
plantarum 1. Alkaline fermentations are carried out by
Bacillus species particularly B. subtilis, which produce a
sticky mucilage on the surface of the beans and strong
proteolytic enzymes which hydrolyse the proteins to
peptides and amino acids. A key reaction is the release
of ammonia which results in a rise in pH from neutral to
pH 8.0 and higher. The relatively high pH in combination
with natural toxicity of ammonia inhibits the contaminants
(Njoku et al., 1990; Sarkar and Tamang, 1994).
Monoculture fermentation of soybeans by B. subtilis has
been reported to produce the best quality kinema, a
traditional food condiment from soybean (Sarkar and
Tamang, 1994). The increase in pH in alkaline
fermenentation might be due to proteolytic activity of the
Bacillus species responsible for the fermentation (Ouoba
et al., 2007). Microbial fermentation of dawadawa has
been found to involve only bacteria since associated
fungi have been regarded as incidental and do not play
any notable role in the fermentation process
(Ikenebomeh and Ingram, 1986).
Fermentation of some plant materials is brought about
by a consortium of microbes and in most cases initiated
by Bacillus species with the assistance of lactic acid
bacteria (Antai and Ibrahim, 1986). Campbell-Platt (1980)
also mentioned lactic acid bacteria (especially species of
Lactobacillus and Pediococcus) as partakers in the
fermentation of dawadawa that gives characteristic
properties to the final product. L. mesenteriodes has
been isolated from fermented dawadawa (Uaboi-Egbemi
et al., 2009) and they also reported that Lactic acid
bacteria from dawadawa may play preservative role in
the final product from African locust beans fermentation.
Mohan and Janardhanan (1995) opined that the
prohibitive cost of animal proteins in developing countries
including Nigeria, calls for extensive exploitation of plant
protein sources, which are economically cheaper. The
result of the microbial study showed that the there is
likelihood of obtaining species that would serve as
Ojinnaka et al.
13
Table 1. Biochemical tests and probable identification of Bacillus isolates.
S/N
TESTS
1
2
3
4
5
6
7
8
9
GLY
ERY
DARA
LARA
RIB
DXYL
LXYL
ADO
MDX
10
11
12
13
14
15
16
17
18
19
20
GAL
GLU
FRU
MNE
SBE
RHA
DUL
INO
MAN
SOR
MDM
21
MDG
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
NAG
AMY
ARB
ESC
SAL
CEL
MAL
LAC
MEL
SAC
TRE
INU
MLZ
RAF
AMD
GLYG
XLT
GEN
TUR
LYX
TAG
DFUC
LFUC
DARL
LARL
GNT
2KG
5KG
Glycerol
Erytrol
D-arabinose
L-arabinose
Ribose
D-Xylose
L-Xilose
D-Adonitol
Metil-βDXylopyranoside
Galactose
Glucose
Fructose
Manose
Sorbose
Rhamnose
Dulcitol
Inozitol
Manitol
Sorbitol
Metil-αDManopyranoside
Metil-ΑdGlucopyranoside
N-Acetylglucosamine
Amygdaline
Arbutine
Esculin
Salicin
D-Celobiose
D-Maltose
D-Lactose
D-Melibiose
D-Zaharose
D-Trehalose
Inuline
D-Melezitose
D-Rafinose
Starch
Glycogen
Xylitol
Gentiobiose
D-Turanose
D-Lyxose
D-Tagatose
D-Fucose
L-Fucose
D-Arabitol
L-Arabitol
Potasium gluconate
2-Ketogluconate
5-Ketogluconate
OGB6:
B.circulans2
+
+
+
-
OGB9:
B.subtilis
+
+
+
+
-
OGB2:
B.megaterium2
+
+
+
+
-
OGB1:B.stearoth
ermophilus
+
-
MO6A:Brevibacillus
brevis
-
+
+
+
+
+
+
-
+
+
+
+
+
-
+
+
+
+
-
+
+
+
-
+
+
-
-
+
-
-
-
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
-
+
+
+
-
technologically useful candidates as starter cultures for
use as biopreservative of the food condiment. So its
incorporation to the fermentation of castor oil bean into
ogiri condiment as mixed starter culture can be studied
having known the various species of Bacillus and LAB
species isolated in this study.
Figure 1 shows the activities of proteinase enzyme during
the fermentation of R. communis into ogiri using B.
14
Sky J. Food Sci.
Table 2. Results of bacteria Identification using API 20E kit for Bacillus species.
S/N
1
2
3
4
5
6
7
8
9
10
11
Tests
ONPG
ADH
LDC
ODC
CIT
H2S
URE
TDA
IND
VP
GEL
12
ONPG
Arginine
Lysine
Ornithine
Citrate
Na thiosulfate
Urea
Tryptophan
Tryptophan
Na pyruvate
Charcoal
gelatin
OGB6:
B. circulans2
+
-
NIT
-
OGB9:
B.subtilis
+
+
+
+
OGB2:
B. megaterium2
+
+
+
OGB1:B.
stearothermophilus
+
MO6A:Brevibacillus
brevis
+
+
+
+
+
+
-
-
-
Table 3. Biochemical tests and probable identification of Bacillus isolates using the API 50 CHL kit.
S/N
Carbohydrate
1
2
3
4
5
6
7
8
9
GLY
ERY
DARA
LARA
RIB
DXYL
LXYL
ADO
MDX
10
11
12
13
14
15
16
17
18
19
20
GAL
GLU
FRU
MNE
SBE
RHA
DUL
INO
MAN
SOR
MDM
21
MDG
22
NAG
23
24
25
26
27
28
29
30
31
32
33
34
35
AMY
ARB
ESC
SAL
CEL
MAL
LAC
MEL
SAC
TRE
INU
MLZ
RAF
Glycerol
Erytrol
D-arabinose
L-arabinose
Ribose
D-Xylose
L-Xilose
D-Adonitol
Metil-βDXylopyranoside
Galactose
Glucose
Fructose
Manose
Sorbose
Rhamnose
Dulcitol
Inozitol
Manitol
Sorbitol
Metil-αDManopyranoside
Metil-ΑdGlucopyranoside
NAcetylglucosamine
Amygdaline
Arbutine
Esculin
Salicin
D-Celobiose
D-Maltose
D-Lactose
D-Melibiose
D-Zaharose
D-Trehalose
Inuline
D-Melezitose
D-Rafinose
OGL7:
L. pentosus
+
+
+
+
-
OGL6:
L. plantarum
+
+
+
+
-
0GL5:
L. plantarum1
+
+
+
-
OGL4:
L. plantarum1
+
+
+
-
OGL3: L. pentosus
+
+
+
+
+
+
-
+
+
+
+
+
+
-
+
+
+
+
+
+
-
+
+
+
+
+
+
-
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
Table 3 cont.
36
37
38
39
40
41
42
43
44
45
46
47
AMD
GLYG
XLT
GEN
TUR
LYX
TAG
DFUC
LFUC
DARL
LARL
GNT
48
49
2KG
5KG
Ojinnaka et al.
Starch
Glycogen
Xylitol
Gentiobiose
D-Turanose
D-Lyxose
D-Tagatose
D-Fucose
L-Fucose
D-Arabitol
L-Arabitol
Potasium
gluconate
2-Ketogluconate
5-Ketogluconate
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
-
-
-
-
15
0% Nacl/lime
2% Nacl
4.5
3% lime
Proteinase activitity (µg/ml)
4
3.5
3
2.5
2
1.5
1
0.5
0
0
24
48
72
96
Ferm entation period (hours)
Figure 1. Proteinase activity during fermentation of castor bean seed into ogiri. LSD0.05 = NS.
subtilis as starter culture. There were gradual increases
in the activities of the proteinase enzyme for 0 – 96 h of
continuous fermentation. Though there were no
significant difference (P>0.05) in the graph of interaction
showing the proteinase enzyme activity in the three
samples. All the samples showed increase in proteinase
activity as fermentation period progressed. Though
sample B1 containing 0%NaCl/Lime increased steadily (
5.69 – 7.01 µg/ml) in the lipase activity compared to the
the other samples B2 (2% NaCl) 2.99 – 4.79 µg/ml and
B3 (3% Lime) 2.22 – 4.92 µg/ml. Proteinase activity has
been reported to be abundant in the fermentation of
similar protein rich foods (Omafuvbe et al., 2002). This is
probably due to a consistently active proteinase activity
resulting in rapid amino acid production. The high
proteinase activity observed in this study may also be
due to the high protein content of R. communis seeds.
Protein in R. communis has been reported to constitute
about 25% or more of the seed (Verscht et al., 2006).
In the fermentation of other protein-rich seeds,
proteinases have also been found to be abundant. This is
true of the fermentation of soybean to produce Japanese
miso, soy-sauce tofu, a Chinese soybean curd, tempe, an
Indonesian soybean fermented food (Odunfa, 1985) and
okpehe, a Nigerian seasoning condiment (Sanni, 1993).
The proteinase enzyme is considered the most
important enzyme in ugba fermentation (Ogueke et al.,
2010). Proteolytic activity has been reported to be
abundant in the fermentation of similar protein rich foods
(Fadahunsi and Sanni, 2010). Detailed studies on the
degradation of African locust bean proteins have been
16
Sky J. Food Sci.
0% Nacl/lime
2% Nacl
9
3% lime
8
Lipase activity
(µg/ml)
7
6
5
4
3
2
1
0
0
24
48
72
96
Ferm entation period (hours)
Figure 2. Lipase activity during fermentation of castor bean seed into ogiri. LSD0.05 = NS.
reported for pure cultures of Bacillus species isolated
from different products of soumbala, a fermented African
locust bean (Ouoba et al., 2003). In a study of the
isolates of B. subtilis and B. pumilus, Ouoba et al. (2003)
demonstrated the different abilities to degrade African
locust bean proteins leading to different profiles of water
soluble proteins and free amino acids and they also
reported that B. subtilis have high proteolytic activity.
Proteolysis has been reported as the main metabolic
activity during the fermentation of African locust bean
which also contributes to the development of texture and
flavour of fermented products (Allagheny et al., 1996;
Ouoba et al., 2003). The desired state of fermentation of
the condiments is indicated by the characteristic
ammonia odour produced as a result of breakdown of
amino acids during fermentation (Omafuvbe, 1998).
Figure 2 shows the lipase activity of fermenting castor
bean mash. There was no significant difference (P>0.05)
between the samples and fermentation time. Though
sample B1 containing 0%NaCl/Lime increased steadily (
5.69 – 7.01 µg/ml) in the lipase activity compared to the
the other samples B2 (2% NaCl) 2.99 – 4.79 µg/ml and
B3 (3% Lime) 2.22 – 4.92 µg/ml. There were little
fluctuations in lipase activities in samples B2 and B3.
Njoku and Okemadu (1989) reported the activity of lipase
as minimal compared to proteinase and amylase
enzymes. This agrees with the report of Achinewhu
(1986) that fermentation has no appreciable effect on the
fatty acid content of P. macrophylla. The minimal activity
of the lipase could be attributed to the effect of NaCl that
is usually added during fermentation. Ogunshe et al.
(2007) reported that there was fluctuation in lipase
activity throughout the fermentation of afiyo, a Nigerian
fermented food condiment from Prosopis africana. They
reported that lipase activities were maximal at days 3 and
5 of fermentation. Odibo et al. (2008) observed very
strong increased lipase activity throughout the duration of
fermentation of seeds of Prosopis africana
for
production of ogiri-okpei. Increase in lipase activity was
also reported by Fadahunsi and Sanni (2010) in tempeh,
fermented bambara groundnut. Inhibition in lipase activity
is reported to be desirable especially during the
production of local condiments because of the problem of
objectionable taste and development of rancidity
(Odunfa, 1985).
Figure 3 shows the changes in amylase activity during
fermentation of castor oil bean into ogiri. There exists a
significant difference (P<0.05) in the interaction of the
samples with fermentation time. The sample containing
0%NaCl/Lime, B1, has higher amylase activity than the
other two samples B2 (2%NaCl) and B3 (3% lime). There
was a steady increase in the amylase activity of sample
B1 while those of B2 and B3 experienced fluctuations in
their amylase activity but increased steadily from 72 h
fermentation. Ogunshe et al, (2007) reported an initial
increase in amylase activity in fermenting afiyo and
subsequent decrease after the first 24h fermentation
while Kpikpi et al. (2009) reported continuous increase in
amylase activity in ‘kantong’ fermentation using Ceiba
pentandra seeds. Njoku and Okemadu (1989) detected
alpha-amylase from the start of ugba fermentation. They
Ojinnaka et al.
17
0% Nacl/lime
2% Nacl
6
3% lime
Amylase activity (µg/ml)
5
4
3
2
1
0
0
24
48
72
96
Ferm entation period (hours)
Figure 3. Amylase activity during fermentation of castor bean seed into ogiri. Vertical bar = LSD0.05.
reported that these enzymes attained their maximum
levels at 24 – 36 h and they suggested that this could be
assumed to be the period of maximum microbial activity.
The initial enzyme activity detected could be due to the
activity of the natural microflora of the oil bean which
developed particularly during the soaking of the cooked
beans. Njoku and Okemadu (1989) therefore suggested
that it could well be that fermentation began much earlier
during the soaking of the sliced beans. Achinewhu (1986)
reported the breakdown of complex carbohydrates into
simple monosaccharide’s during ugba fermentation while
Mbajunwa et al. (1998) observed that Bacillus subtilis
was the main fermenting microorganism. Bacillus species
are found to be important producers of alpha-amylases
(Enujiugha et al., 2002). Afolabi et al. (2011) in their study
of fermented papaya seeds observed low alpha-amylase
activity as an indication of its low starch content and
reported that fermentation process will not affect the
qualities (viscosity and texture) of the final products
obtained from the seed.
Conclusion
Using High Performance Liquid Chromatography (HPLC)
to monitor enzymes during 96 h fermentation of castor oil
bean into ogiri goes to show the in-depth molecular
study of the changes occurring during castor bean
fermentation as well as make appropriate modification
where applicable. However, the production of ogiri from
castor oil bean offers a method to utilize castor oil bean
as a condiment otherwise it is inedible due to its toxic
constituent, ricin.
Proteinase and lipase activities are reduced by
inclusion of additives but not to significant (P>0.05)
levels. Amylase activity which is indicative of
carbohydrate utilization is reduced in the presence of 2%
NaCl as well as 3% lime suggesting that carbohydrate
are the major nutrients utilized in the absence of these
additives.
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