Professional Documents
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Journal of Engineering & Applied Sciences Vol. 2(1), pp10-15, July, 2013
International Research Journal Group
All Rights Reserved
Available online http://www.irjset.com
Department of Microbiology, Ibrahim Badamasi Babangida University, Lapai, Niger State, Nigeria
Department of Biochemistry, Ibrahim Badamasi Babangida University, Lapai, Niger State, Nigeria
3
Department of Crop Production, Ibrahim Badamasi Babangida University, Lapai, Niger State, Nigeria
4
Department of Microbiology, Federal University of Technology, Minna, Niger State, Nigeria
2
Abstract The influence of nutrient supplements on bacteriocin production by Lactobacillus bulgaricus, Y34 and Lactococ-
cus lactis N22 was carried out. The supplements of nutrients with De Man Rogosa Sharpe (M.R.S) broth demonstrated that
large quantities of bacteriocins could be produce by addition of glucose (1%), Nacl (1 and 2%), beef extract (2%), yeast extract (1, 2 and 3%), tween 80 (0.5 and 1%) while the addition of 0.1% and 0.5% triammonum citrate, 0.5 and 1% sodium acetate, magnesium sulphate, manganese sulphate and potassium phosphate had effects on the bacteriocin production. Bacteriocin
produced by L. bulgaricus. Y34 and L. lactis N22 had large spectrum of inhibition against the pathogenic, food spoilage microorganisms (indicator microorganisms) employed as test strains. The bacteriocins produced by test strains inhibited the
growth of indicator microorganisms i.e Staphylococcus aureus N23, Salmonella typhimurium N17, Bacillus cereus N12,
Shigella dysentriae N25, and Pseudomonas aeruginosa N1. L. bulgaricus Y34 had the highest bacteriocin activity of
600000.0 AU/mL while L. lactis N22 had 580000.0 AU/mlL as it highest bacteriocin activity. Maximal bacteriocin production was achieved at pH between 3.9 and 4.5 and incubation period of 48 hours at temperature of 30oC for the two species of
lactic acid bacteria used in this study.
Keywords: Bacteriocins, Lactic acid bacteria, De Man Rogosa Sharpe broth, indicator microorganisms, Nutrient supplements.
* Corresponding author:
mosada78@yahoo.com (Mohammed,S.S.D)
Published online at http:/www.irjset.com
Copyright 2013 International Research Journal Group
1. Introduction
3. Results
Quantity of
Nutrient used (%)
Growth
(580nm)
pH of
medium
Bacteriocin
activity (Au/ml)
MRS (unsupplemented)
0.0
0.90
3.91
5800 0.00*
MRS + glucose
1.0
0.90
4.07
6000 0.00*
2.0
0.92
4.09
3000 0.00
1.0
0.70
3.92
6000 0.00*
2.0
0.73
3.97
5800 0.00*
3.0
0.60
3.95
3000 0.00
1.0
0.90
3.93
3000 0.00
2.0
0.92
3.98
6000 0.00*
1.0
0.92
4.03
5800 0.00*
2.0
0.90
4.07
6000 0.00*
3.0
0.89
4.20
6000 0.00*
0.5
0.70
4.01
5800 0.00*
1.0
0.71
4.00
6000 0.00*
MRS + Triammonium
0.1
0.40
4.01
3000 0.00
Citrate
0.5
0.70
MRS + Nacl
MRS + Tween 80
MRS + MnSo4.4H2
MRS + K2 HPO4
MRS + Sodium acetate
3000 0.00
0.1
0.89
4.93
3000 0.00
0.5
0.70
4.00
3000 0.00
0.1
0.50
4.01
3000 0.00
0.5
0.80
0.04
3000 0.00
0.50
3.98
3000 0.00
0.60
4.01
3000 0.00
4.06
3000 0.00
0.5
1.0
4.06
0.5
1.0
0.90
0.60
4.00
3000 0.00
Growth, pH and bacteriocin determinations were done after 48 hours of incubation. *:potential bacteriocins produced.
MRS: Deman Rogosa Sharpe medium; AU/mL: Activity per milliliter; nm: nanometer Y: Yoghurt
Table 2. Effects of nutrient Supplements on bacteriocin production by Lactococcus lactis N 22
Medium composition
Quantity of
Nutrient used (%)
Growth
pH of
(580nm)
medium
Bacteriocin
activity (Au/ml)
MRS (unsupplemented)
0.0
0.90
3.72
5600 0.00*
MRS + glucose
1.0
0.80
4.03
5800 0.00*
2.0
0.72
4.07
2900 0.00
1.0
0.70
3.84
5800 0.00*
2.0
0.70
3.90
5600 0.00*
3.0
0.60
3.95
2900 0.00
1.0
0.90
3.93
5800 0.00*
MRS + Nacl
2.0
MRS + yeast extract
0.89
3.90
5800 0.00*
1.0
0.94
3.03
5800 0.00*
2.0
0.90
4.07
5800 0.00*
3.0
0.90
4.50
5800 0.00*
0.5
0.70
4.02
5600 0.00*
1.0
0.60
3.09
5800 0.00*
MRS + Triammonium
0.1
0.40
3.07
5600 0.00*
Citrate
0.5
0.70
4.06
2900 0.00
MRS + MnSO4.4H2O
0.1
0.89
4.90
2900 0.00
MRS + Tween 80
0.5
0.80
4.00
2900 0.00
MRS + K2 HPO4
0.1
0.70
3.09
2900 0.00
0.5
0.90
4.06
2900 0.00
0.5
0.50
3.90
1.0
0.60
4.01
2900 0.00
0.5
0.90
3.06
2900 0.00
1.0
0.60
4.00
2900 0.00
2900 0.00
Growth, pH and bacteriocin determinations were done after 48 hours of incubation. *:potential bacteriocins produced.
MRS: Deman Rogosa Sharpe medium; AU/mL: Activity per milliliter; nm: nanometer Y: Yoghurt
Table 3 Inhibition of indicator microorganisms by bacteriocins produced by Lactobacillus bulgaricus Y34 and Lactococcus lactis N22 after nutrient Supplements of MRS
Indicator microorganisms
Source
Nono
+(6mm)
+(6mm)
Nono
+(6mm)
+(6mm)
Bacillus cereus N 12
Nono
+(6mm)
+(6mm)
Shigella dysenteriae N 25
Nono
+(6mm)
+(6mm)
Pseudomonas aeruginosa N1
Nono
+(6mm)
+(6mm)
Key:
+:Inhibition
-:No Inhibition
N: Nono
4. Discussion
The present study was aimed at determining the influence of
nutrient supplements on bacteriocin production. L. burglarious Y34 and L. lactis N22 were able to produce bacteriocin
which had wide inhibitory spectrum towards the indicator
microorganisms. Thus, Variation in the concentration of constituents/supplements of cultivating media (MRS broth) has
an influence on the amount of bacteriocin produced by L.
burglaricus Y34 and L. lactis N22. The highest bacteriocin
activity of 6000+0.00 AU/mL was exhibited by L. burglarious Y34 when 2%. Glucose, 1% Nacl, 2% beef extract,1, 2
and 3% yeast extract and 1% tween 80 was added to MRS
broth. While L lactis N22 exhibited highest bacteriocin activity of 58000.00 AU/mL when 2% beef extract, 1, 2 and 3%
yeast extract, 0.5 and 1% tween 80 were added to MRS broth.
While the supplements of triammonium citrate, manganese
sulphate, potassium hydrogen sulphate, sodium acetate and
magnesium sulphate at 0.1% and 0.5% yielded no improvement in bacteriocin production. Similar observations have
been made previously by Daba et al. (1993) who reported
similar findings in the production of mensenterocin 5. Sani et
al. (1999) also reported that highest bacteriocin activity was
obtained when glucose and peptone were varied to 0.25%
and 0.5% in the constitute MRS broth. While bacteriocin
activity was not detected at 2% glucose and peptone level.
The ineffectiveness of manganese sulphate, potassium hydrogen sulphate, sodium acetate and magnesium sulphate
supplemented with the MRS broth might be due to the activity of extracellular endogenous proteinases induced during
the growth phase of the test strains. This is similar to the
findings of Paired et al. (1990) who revealed that the decrease in bacteriocin production by Lactococcus lactis CNRZ
481 after nutrient supplementation could be due to activity of
extracellular proteinases induced during the growth phase.
Some reports indicate that bacteriocin are produced throughout the experimental growth phase and note solely during late
logarithmic or early stationary phase (Joerger and Klaenhammer, 1986., Paid et al., 1990). The bacteriocin produce
by L bulgaricus Y34 exhibited bacteriocin activity between
3000+0.00 to 6000+0.00 Au/ml while the bacteriocin produced by L. lactis N22 exhibited histrionic activity between
29000.00 to 58000.00 Au/ml against Staphylococcus
aureus N23, Salmonella typhimurium N17, Bacillus cereus
N12, Shigella dysenteriae N25 and Pseudomonas aeruginosa
N1. This is in conformity with the findings of Ogumbanwo
et al. (2003) who reported the bacteriocin produced by Lactobacillus plantarium F1, and Lactobacillus brevis OG1 exhibited bacteriocin activity between 3200+0.00 to 6400+0.00
Au/ml against Escherichia coli, Listeria monocytogene and
Enterococcus faecalis. This findings also agrees with Mohammed et al.( 2012 ) who reported that pediocin and nisin
produced by Pediococcus halophilus W9 and Lactococcus
lactis N24 exhibited bacteriocin activity between 50000.00
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