scholarly journals Production and characterization of bacteriocin produced by lactic acid bacteria isolated from rusip

Author(s):  
Arifah Kusumarwati ◽  
Ninoek Indriati ◽  
Irma Hermana

Research was conducted to produce and characterize bacteriocin produced by lactic acid bacteria (LAB) isolated from rusip, a traditional Bangkanese fermented fish product. Experiment was started by isolation of lactic acid bacteria from rusip, followed by screening to obtain the best isolate which has the highest bacterial inhibition activity. The selected isolate was then identified and used to produce crude bacteriocin. The crude bacteriocin was characterized through its stability in high temperature and proteolytic enzymes, inhibitory spectrum, pH sensitivity and effect of surfactants. The result showed that CN1.10a isolate which was identified as Lactococcus lactis subsp lactis has the highest bacterial inhibition activities; therefore it was selected to produce crude bacteriocin. The bacteriocin produced was heat stable, sensitive to proteolytic enzymes i.e. proteinase-K and papain but not to RNase. It inhibited Escherichia coli, Listeria monocytogenes and Lactobacillus plantarum. It stable at pH 2.0 to 6.0. Among surfactans used sodium dodecyl sulphate (SDS), lauryl sarcosine and EDTA were able to stimulate bacteriocin production, while the production were strongly inhibited by Tween 20, Tween 80, Triton X-100 and urea. Based on the above characteristic, the bacteriocin was suitable to be used as a preservative of food which has to be processed at high temperature.

2007 ◽  
Vol 73 (7) ◽  
pp. 2247-2250 ◽  
Author(s):  
Sirinat Srionnual ◽  
Fujitoshi Yanagida ◽  
Li-Hsiu Lin ◽  
Kuang-Nan Hsiao ◽  
Yi-sheng Chen

ABSTRACT Weissella cibaria 110, isolated from the Thai fermented fish product plaa-som, was found to produce a bacteriocin active against some gram-positive bacteria. Bacteriocin activity was not eliminated by exposure to high temperatures or catalase but was destroyed by exposure to the proteolytic enzymes proteinase K and trypsin. The bacteriocin from W. cibaria 110 was purified, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the purified bacteriocin contained one protein band that was approximately 2.5 kDa in size. Mass spectrometry analysis showed the mass of the peptide to be approximately 3,487.8 Da. N-terminal amino acid sequence analysis was performed, and 27 amino acids were identified. Because it has no similarity to other known bacteriocins, this bacteriocin was defined as a new bacteriocin and termed weissellicin 110.


2015 ◽  
Vol 83 (1) ◽  
pp. 115-124 ◽  
Author(s):  
Fabricio L Tulini ◽  
Nolwenn Hymery ◽  
Thomas Haertlé ◽  
Gwenaelle Le Blay ◽  
Elaine C P De Martinis

Lactic acid bacteria (LAB) can be isolated from different sources such as milk and cheese, and the lipolytic, proteolytic and glycolytic enzymes of LAB are important in cheese preservation and in flavour production. Moreover, LAB produce several antimicrobial compounds which make these bacteria interesting for food biopreservation. These characteristics stimulate the search of new strains with technological potential. From 156 milk and cheese samples from cow, buffalo and goat, 815 isolates were obtained on selective agars for LAB. Pure cultures were evaluated for antimicrobial activities by agar antagonism tests and for proteolytic activity on milk proteins by cultivation on agar plates. The most proteolytic isolates were also tested by cultivation in skim milk followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the fermented milk. Among the 815 tested isolates, three of them identified asStreptococcus uberis(strains FT86, FT126 and FT190) were bacteriocin producers, whereas four other ones identified asWeissella confusaFT424,W. hellenicaFT476,Leuconostoc citreumFT671 andLactobacillus plantarumFT723 showed high antifungal activity in preliminary assays. Complementary analyses showed that the most antifungal strain wasL. plantarumFT723 that inhibitedPenicillium expansumin modified MRS agar (De Man, Rogosa, Sharpe, without acetate) and fermented milk model, however no inhibition was observed againstYarrowia lipolytica. The proteolytic capacities of three highly proteolytic isolates identified asEnterococcus faecalis(strains FT132 and FT522) andLactobacillus paracaseiFT700 were confirmed by SDS–PAGE, as visualized by the digestion of caseins and whey proteins (β-lactoglobulin and α-lactalbumin). These results suggest potential applications of these isolates or their activities (proteolytic activity or production of antimicrobials) in dairy foods production.


2010 ◽  
Vol 27 (Special Issue 2) ◽  
pp. 12-17
Author(s):  
K. Kučerová ◽  
I. Korbová ◽  
Š. Horáčková ◽  
E. Šviráková ◽  
M. Plocková

A collection of lactic acid bacteria (38 Enterococcus and 41 Lactobacillus strains) was tested for the antilisterial activity against 15 Listeria spp. strains (two L. monocytogenes, one L. ivanovii and 12 L. innocua strains) using agar spot method. Out of all 79 bacteria only six Enterococcus strains (1/3A, 3/3A, 6/4D, 6/1A, 1282 and EN3) exhibited antilisterial activity against almost all used indicator strains, when their live cells were tested. When their cell free neutralised supernatants (CFNS) were tested against four selected indicator strains (L. innocua Ln-03, Ln-06, Ln-10 and L. monocytogenes CCM5576) only two Enterococcus spp. strains were active – E. faecalis 6/1A strain from raw cow milk of minor interest due to the activity of its CFNS only against L. innocua Ln-06 and thermolability of the compound and E. mundtii 1282 strain from goat raw milk with CFNS active against 13 Listeria spp. strains including L. monocytogenes. E. mundtii 1282 strain produced probably a bacteriocin, because it completely lost the activity after treatment CFNS with proteinase K.


2006 ◽  
Vol 69 (8) ◽  
pp. 1937-1943 ◽  
Author(s):  
PONGSAK RATTANACHAIKUNSOPON ◽  
PARICHAT PHUMKHACHORN

Lactobacillus plantarum N014 was isolated from nham, a traditional Thai fermented pork, and exhibited antimicrobial activity against Listeria monocytogenes. Its bacteriocin had a broad inhibitory spectrum toward both gram-positive and gram-negative bacteria. The bacteriocin activity was sensitive to all proteolytic enzymes used in this study, including papain, pepsin, pronase E, proteinase K, and trypsin, but was resistant to the other enzymes, such as α-amylase, lipase A, and lysozyme. Furthermore, activity was stable over various heat treatments and pH values. The bacteriocin exerted a bacteriolytic mode of action. It was produced during the exponential growth phase and reached its highest level as producer cells entered the stationary phase. Adsorption of the bacteriocin onto producer cells was pH-dependent. No bacteriocin adsorption was detected at pH 1 to 3, whereas 100% bacteriocin adsorption was found at pH 7. Plasmid isolation revealed that L. plantarum N014 contained no plasmids. From Tricine–sodium dodecyl sulfate–polyacrylamide gel electrophoresis and growth inhibition testing against L. monocytogenes, the estimated molecular mass of L. plantarum N014 bacteriocin was 8 kDa.


2012 ◽  
Vol 57 (No. 3) ◽  
pp. 137-149 ◽  
Author(s):  
H. Musikasang ◽  
N. Sohsomboon ◽  
A. Tani ◽  
S. Maneerat

Bacteriocin-producing lactic acid bacteria (LAB) were isolated and screened from the gastrointestinal tract (GIT) of Thai indigenous chickens. The bacteriocinogenic activities and the primary probiotic properties were determined. The bacteriocins produced by 14 strains of selected LAB displayed inhibitory activity against indicator strains after the supernatants were neutralized with NaOH in the following species: Lactobacillus sakei subsp. sakei JCM1157, Enterococcus faecalis VanB, Bacillus sp., and Listeria monocytogenes. The antagonistic acti-vity of selected LAB was inactivated or decreased after being treated with proteolytic enzymes (α-chymotrypsin and trypsin). CR5-1 strain exhibited the highest level of activity (5120 AU/ml) in the stationary phase against L. sakei subsp. sakei JCM1157 in MRS broth at 37°C. The nine isolates of selected LAB were investigated for primary probiotic properties. The survival of the nine isolates was found to decrease approximately by 3 log CFU/ml after passing through the gastrointestinal conditions. All isolates exhibited protein digestion on agar plates but no isolates showed the ability to digest starch and lipid. Most of them showed high susceptibilities to some antibiotics (penicillin G, tetracycline and erythromycin). Thirteen LAB strains producing bacteriocin with strongly inhibitory activity were identified as Lactobacillus salivarius and only one strain was identified by 16S rDNA sequence analysis as Lactobacillus agilis.    


LWT ◽  
1996 ◽  
Vol 29 (5-6) ◽  
pp. 561-569 ◽  
Author(s):  
Marco Gobbetti ◽  
Emanuele Smacchi ◽  
Patrick Fox ◽  
Leszek Stepaniak ◽  
Aldo Corsetti

2012 ◽  
Vol 3 (4) ◽  
pp. 319-330 ◽  
Author(s):  
S.D. Todorov ◽  
L. Favaro ◽  
P. Gibbs ◽  
M. Vaz-Velho

Strain ST211CH, identified as a strain of Enterococcus faecium, isolated from Lombo produced a bacteriocin that inhibited the growth of Enterococcus spp., Listeria spp., Klebsiella spp., Lactobacillus spp., Pseudomonas spp., Staphylococcus spp. and Streptococcus spp. The mode of action of the bacteriocin named as bacteriocin ST211Ch was bactericidal against Enterococcus faecalis ATCC19443. As determined by Tricine-SDS-PAGE, the approximate molecular mass of the bacteriocin was 8.0 kDa. Loss in antimicrobial activity was recorded after treatment with proteolytic enzymes. Maximum activity of bacteriocin ST211Ch was measured in broth cultures of E. faecium strain ST211Ch after 24 h; thereafter, the activity was reduced. Bacteriocin ST211Ch remained active after exposure to various temperatures and pHs, as well as to Triton X-100, Tween-80, Tween-20, sodium dodecyl sulfate, NaCl, urea and EDTA. Effect of media components on production of bacteriocin ST211Ch was also studied. On the basis of PCR reactions targeting different bacteriocin genes, i.e. enterocins, curvacins and sakacins, no evidences for the presence of these genes in the total DNA of E. faecium strain ST211Ch was obtained. The bacterium most probably produced a bacteriocin different from those mentioned above. Based on the antimicrobial spectrum, stability and mode of action of bacteriocin ST211CH, E. faecium strain ST211Ch might be considered as a potential candidate with beneficial properties for use in biopreservation to control food spoilage bacteria.


2021 ◽  
Vol 2 (3) ◽  
pp. 170-178
Author(s):  
Tita Rialita ◽  
Sumanti Debby Moody ◽  
Edy Subroto ◽  
Heditia Febby Susanto

Bacteriocin was bacterial metabolite that have antimicrobial properties, so it had the potential to be used as food bio preservatives. Bacteriocin was produced by lactic acid bacteria (LAB), one of the sources of which was from smoked fish products. Some regions in Indonesia produce various types of smoked fish from various types of fish, which were thought to contain bacteriocin-producing lactic acid; one of them was giant cathfish (Arius thalassinus). This study aims to obtain LAB isolates that have strong antimicrobial activity and have the potential to produce bacteriocin-like from smoked giant catfish (Arius thalassinus).  The research method used an experimental method that analyzed descriptively.  Based on the results, there were 15 isolates LAB isolated from smoked giant catfish. Three selected isolates showed strong antimicrobial activity inhibiting E. coli and S. aureus bacteria, and the most effective inhibiting Salmonella sp. One selected LAB isolates identified Pediococcus acidilactici suspected to produce pediocin bacteriocin-like, while the other two isolates identified Lactobacillus plantarum sp 1 and Lactobacillus plantarum sp 2 which suspected to produce plantaricin bacteriocin. Bacteriocin from the three isolates of LAB had characteristics stable to temperatures up to 121oC, stable in pH range 2-6, and bacteriocin activity increased with the addition of SDS (Sodium Dodecyl Sulfate) and EDTA (Ethylenediaminetetraacetic acid) surfactants. The conclusion was that the bacteriocin produced was stable at high temperature, low pH, and resistence in the presence of surfactants, so it had the potential to be developed as biopreservatives material in preserving fish-based foods


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