Tuberculosis(TB) is a communicable disease caused by Mycobacterium tuberculosis(M. tuberculosis). WHO estimated that 10.4 million new(incident) TB cases worldwide in year 2016. The increased prevalence of drug resista...Tuberculosis(TB) is a communicable disease caused by Mycobacterium tuberculosis(M. tuberculosis). WHO estimated that 10.4 million new(incident) TB cases worldwide in year 2016. The increased prevalence of drug resistant strains and side effects associated with the current anti-tubercular drugs make the treatment options more complicated. Hence, there are necessities to identify new drug candidates to fight against various sub-populations of M. tuberculosis with less or no toxicity/side effects and shorter treatment duration. Bacteriocins produced by lactic acid bacteria(LAB) attract attention of researchers because of its "Generally recognized as safe" status. LAB and its bacteriocins possess an effective antimicrobial activity against various bacteria and fungi. Interestingly bacteriocins such as nisin and lacticin 3147 have shown antimycobacterial activity in vitro. As probiotics, LAB plays a vital role in promoting various health benefits including ability to modulate immune response against various infectious diseases. LAB and its metabolic products activate immune system and thereby limiting the M. tuberculosis pathogenesis. The protein and peptide engineering techniques paved the ways to obtain hybrid bacteriocin derivatives from the known peptide sequence of existing bacteriocin. In this review, we focus on the antimycobacterial property and immunomodulatory role of LAB and its metabolic products. Techniques for large scale synthesis of potential bacteriocin with multifunctional activity and enhanced stability are also discussed.展开更多
Objective:To screen the bacteriocinogenic isolate from buffalo milk and to characterize it on physical,chemical and biological aspects for the application in biopreservation.Methods:Bacillus cereus(B.cereus)was isolat...Objective:To screen the bacteriocinogenic isolate from buffalo milk and to characterize it on physical,chemical and biological aspects for the application in biopreservation.Methods:Bacillus cereus(B.cereus)was isolated and assessed for its baceteriocinogenic activity.Bacteriocin was produced and purified by ammonium sulphate precipitation,dialysis and gel filtration chromatography.Purified bacteriocin was used to clieck its antimicrobial activity against food borne bacteria.Effect and stability of bacteripcin was determined with the respect to temperature,pH,enzymes,organic solvents and chemicals.Bacteriocin was also subjected to SDS PAGE analysis to determine its molecular weight.In addition,functional groups exist in the bacteriocin was determined by FTIR analysis.Results:B.cereus was identified by 16S rRNA sequence analysis.Bacteriocin showed increased activity against all the bacteria used and its activity unit was found to be 51,200 AU/mL.It was stable to high temperature(100℃)and wide range of pH(3-10),sensitive to proteolytic enzymes and resistant to nonprotcolytic enzymes.It was low molecular weight(3.5-6 kDa)protein and FTIR study revealed the presence of amide group and NH stretching,Conclusions:Bacteriocin produced in this study possesses the highest antimicrobial activity against both gram positive and gram negative bacteria thereby it has immense application as biopreservative agent.FTIR proved its peptide nature.展开更多
Biopreservation systems in foods are of increasing interest for industry and consumers. Bacteriocin producing Lactobacillus spp. is considered Generally Recognize as Safe (GRAS), useful to control the fast development...Biopreservation systems in foods are of increasing interest for industry and consumers. Bacteriocin producing Lactobacillus spp. is considered Generally Recognize as Safe (GRAS), useful to control the fast development of pathogens and spoiling microbes in food and feed. Lactobacillus spp. was isolated from traditional winter fermented vegetable cucumber & carrot by the use of selective media. Especially De Man, Rogosa and Sharpe (MRS) Agar media were used to isolate the Lactobacillus species. Morphologically identified by gram staining & colony morphology. Biochemically recognized by catalase, oxidase, MRVP & carbohydrate fermentation test. Antimicrobial activity of Lactobacillus spp. was confirmed by Well Diffusion Method. Molecular characterization of bacteriocin protein and molecular weight determined by SDS PAGE method. The isolate was found to be facultative anaerobic, Gram positive, and catalase negative. The result of antimicrobial activity measured by the Arbitrary Unit (AU/ml) of zone of the inhibition. Six isolates found from the sample but most activities exhibited isolate 4 against Bacillus megaterium (55 mm) zone of diameter. The molecular weight of the washed bacteriocin was calculated to be about 40 kDa (Isolate 1) and 15 kDa & 30 kDa (Isolate 5). Bacteriocin protein reduces chemical preservatives and uses in future as biopreservative in food industry.展开更多
Lactobacilli belong to the group of lactic acid bacteria (LAB), that have several distinguished abilities such as production of lactic acid, enzymes such as β-Galactosidase and natural antimicrobial substances called...Lactobacilli belong to the group of lactic acid bacteria (LAB), that have several distinguished abilities such as production of lactic acid, enzymes such as β-Galactosidase and natural antimicrobial substances called bacteriocins. Bacteriocin is a biopreservative agent potential of suppressing growth of some contaminant bacteria in food industry but its commercial availability is limited and costly. The study aimed to select isolates of Lactobacillus spp. potential for producing bacteriocins to suppress the growth of Escherichia coli ATCC 25922 and Bacillus subtilis NCIB3610, and to optimize the process of bacteriocin production. Results obtained in this study showed that L. acidophilus isolate CH1 was selected as the best candidate for bacteriocin among the four isolates that tested. The largest amounts of the bacteriocins were synthesized only in MRS medium was supplemented with K2HPO4 (1.0%), Tween 80 (1%), Beef extract (1%), glucose, cyctein and peptone extract (1%). The optimization of culture conditions for bacteriocin production areas showed that corn steep liquor medium was the best medium for all isolates against Bacillus subtilis while no effect was observed on Escherichia coli ATCC 25922 except when used MRS medium. The optimum conditions for bacteriocin production were pH 6.0, temperature 34?C with 4% Phenyl acetamide showing the greatest growth inhibition areas.展开更多
A bacteriocin producing strain of Lactococcus lactis subsp. lactis LABW4 was isolated from naturally fermented milk product which exhibited strong antibacterial activity against Listeria monocytogenes MTCC657, a food ...A bacteriocin producing strain of Lactococcus lactis subsp. lactis LABW4 was isolated from naturally fermented milk product which exhibited strong antibacterial activity against Listeria monocytogenes MTCC657, a food spoilage psychrophilic organism. Both cell free and heat killed supernatants of LABW4 were effective to produce zones of inhibition against L. monocytogenes in vitro. The antibacterial metabolite(s) of LABW4 showed strong cidal effect on the growth of L. monocytogenes. Meat samples, mixed with heat killed supernatant of LABW4 when inoculated with Listeria, remain fresh up to 25 days in refrigerated condition whereas spoilage started immediately after 24 hours of inoculation for control sets. Enhancement of Lactate dehydrogenase of L. monocytogenes upon treatment with LABW4 cell free supernatant suggested its lytic mode of action. Cell lysis or degradations were also supported by scanning electron micrograph of treated cells.展开更多
A collection of lactic acid bacteria isolates from fish viscera was studied and investigated regarding to their functional properties and safety aspects. From these, three isolates GM1, GM2 and GM3 were identified as ...A collection of lactic acid bacteria isolates from fish viscera was studied and investigated regarding to their functional properties and safety aspects. From these, three isolates GM1, GM2 and GM3 were identified as Enterococcus feacium species using molecular methods. Partial Amplified rDNA Restriction Analysis (partial ARDRA) with restriction enzyme HaeIII separated these isolates into distinctive group which suggest genotypic variability within enterococci strains isolated from fish viscera. The three strains GM1, GM2 and GM3 exhibited antimicrobial activity. Indeed strains have been shown to produce bacteriocins with inhibitory effect against food spoilage bacteria and pathogenic fish including Carnobacterium maltaromaticum. The molecular mass of bacteriocin, as calculated by tricine-SDS-PAGE, was found to be 4.5 kDa. All isolates were tested positive upon PCR amplification of enterocin A structural gene. Investigations of antibiotic resistance show that the isolates were mostly sensitive to several antibiotics (ampicillin, penicillin, tetracycline, gentamycine) and resistance to rifampicin. All isolates grow in esculin azide agar as a selective medium for enumeration of probiotic enterococci. This study suggests that our strains can be employed as probiotic or to improve the safety of food products.展开更多
Two lactic acid bacteria strains (At1BEAE22 and At344E21) isolated during tchapalo production were identified on the basis of phenotypic analyses. Bacteriocins produced by these strains were tested for their antimicro...Two lactic acid bacteria strains (At1BEAE22 and At344E21) isolated during tchapalo production were identified on the basis of phenotypic analyses. Bacteriocins produced by these strains were tested for their antimicrobial activities using well diffusion agar method. Heat resistance, pH sensitivity and enzyme treatments were also analyzed. Results showed that both lactic acid bacteria strains were identified as Pediococcus acidilactici. Their bacteriocins inhibited growth of Lactobacillus delbrueckii F/31, Listeria innocua ATCC 33090, Enterococcus faecalis, Enterococcus faecalis ATCC 29212, Streptococcus sp, Enterococcus faecalis CIP 105042 and Enterococcus faecium ATCC 51558. These bacteriocins were heat stable at 60°C for 30 min for all indicator bacteria. However, they remained active only against Lactobacillus delbrueckii and Listeria innocua at 121°C for 60 min. Moreover, they were active in a wide range of pH (3 to 9) with a maximum activity observed at pH 5 and 6 on all indicator bacteria. But, bacteriocin from Pediococcus acidilactici At34E21 was more stable at acidic pH than basic one. The fact that the bacteriocin was inactivated by proteinase K and α-chymotrypsin indicated its proteinaceous nature, a general characteristics of bacteriocins.展开更多
Bacteriocins are a large group of chromosome or plasmid-encoded and ribosomally synthesized low-molecular-weight (2 to 6 kDa) antimicrobial and amphiphilous peptides produced by Gr+ or Gr- bacteria [1]. Their low toxi...Bacteriocins are a large group of chromosome or plasmid-encoded and ribosomally synthesized low-molecular-weight (2 to 6 kDa) antimicrobial and amphiphilous peptides produced by Gr+ or Gr- bacteria [1]. Their low toxicity as well as the absence of allergenicity and reactogenicity is confirmed by testing selected bacteriocins [2] [3]. Bacteriocins can be widely used as preservatives and antibiotic alternatives in medicine. Nisin, a Streptococcus lactis-derived bacteriocin, has been in practice in food industry for a long time. A relevant product contains about 2.5% of nisin. For medical use (e.g., when injected into the blood stream), highly purified drugs are required. However, the yield of bacteriocins accounts for no more than a few percents from the total activity in the culture liquid. In this paper, we propose methods (by example of two B. subtilis strains), allowing to increase the yield up to ~80%. It is believed that other bacteriocins may be purified by these methods and with the same yield.展开更多
Objective: To characterize the bacteriocin-like inhibitory-substances(BLIS) activity of Streptococcus macedonicus MBF10-2 [named BLIS(MBF10-2)], a bacteriocinogenic strain isolated from an Indonesian tofu byproduct.Me...Objective: To characterize the bacteriocin-like inhibitory-substances(BLIS) activity of Streptococcus macedonicus MBF10-2 [named BLIS(MBF10-2)], a bacteriocinogenic strain isolated from an Indonesian tofu byproduct.Methods: BLIS(MBF10-2) was obtained by culturing the bacterium, and standard deferred antagonism assays were used to demonstrate its activity. The antibacterial testing of fractions collected by filtration using 3–30 kDa cut-off membrane sizes were carried out by performing well diffusion method.Results: The growth of Micrococcus luteus, Streptococcus pyogenes, Lactococcus lactis,Leuconostoc mesenteroides and Weissella confusa were inhibited by BLIS(MBF10-2).Interestingly, BLIS-containing fractions obtained from sequential application on ultrafiltration membranes indicated that this bacterium Streptococcus macedonicus MBF10-2 could produces at least two antimicrobial peptides activities, one of which is likely to be a lantibiotic peptide. Potential synergistic activity against certain Gram-positive(but not Gram-negative) species when partnered with antibiotics(ampicillin, tetracycline or kanamycin) were observed.Conclusion: Combination of some BLIS(MBF10-2) active fractions with antibiotics(ampicillin, tetracycline or kanamycin) could demonstrate synergistic activities against certain Gram-positive species.展开更多
This study was performed to determine the effects of bacteriocin-producing and non-bacteriocin-producing Pediococcus acidilactici strains on the immune system and intestinal flora of normal mice.Two P.acidilactici str...This study was performed to determine the effects of bacteriocin-producing and non-bacteriocin-producing Pediococcus acidilactici strains on the immune system and intestinal flora of normal mice.Two P.acidilactici strains with antibacterial activity(P.acidilactici CCFM28 and CCFM18)were obtained based on the inhibition-zone assay.The produced components were identified as bacteriocins through protease treatment,pH adjustment and hydrogen peroxide treatment.Bacteriocin-producing and non-bacteriocin-producing P.acidilactici strains(P.acidilactici CCFM28,CCFM18 and NT17-3)caused significant changes in serum immune factors and intestinal flora of normal mice.After 14 days of intervention,the relative abundance of Firmicutes was significantly decreased,but that of Proteobacteria was significantly increased at the phylum level.At the genus level,the administration of three P.acidilactici strains resulted in the downregulation of Blautia and the upregulation of Ruminococcus and Lactobacillus.Furthermore,there were also different regulations on some probiotic strains,such as Bifidobacterium,Coprococcus and Akkermansia,which were closely related to the antibacterial ability of the bacteriocin and the type of strain.The results indicated that the intervention of different P.acidilactici strains could differently change the structure of intestinal flora in normal mice,which provided theoretical guidance for the selective use of bacteriocin-producing strains for health regulation in the future.展开更多
The use of antimicrobial peptides(AMPs)synthesized by bacteria(bacteriocins)is an alternative for combating multidrug resistant bacterial strains and their production by recombinant route is a viable option for their ...The use of antimicrobial peptides(AMPs)synthesized by bacteria(bacteriocins)is an alternative for combating multidrug resistant bacterial strains and their production by recombinant route is a viable option for their mass production.The bacteriocin E-760 isolated from the genus Enterococcus sp.has been shown to possess inhibitory activity against Gram-negative and Gram-positive bacteria.In this study,the expression of a chimeric protein coding for E-760 in the nucleus of C.reinhardtii was evaluated,as well as,its antibacterial activity.The synthetic gene E-760S was inserted into the genome of C.reinhardtii using Agrobacterium tumefaciens.A transgenic line was identified in TAP medium with hygromycin and also by PCR.The increment in the culture medium temperature of the transgenic strain at 35°C for 10 minutes,increased the production level of the recombinant protein from 0.14(Noninduced culture,NIC)to 0.36%(Induced culture,IC)of total soluble proteins(TSP);this was quantified by an ELISA assay.Recombinant E-760 possesses activity against Staphylococcus aureus in 0.34 U log,Streptococcus agalactiae in 0.48 U log,Enterococcus faecium in 0.36 U log,Pseudomonas aeruginosa in 2 U log and for Klebsiella pneumoniae,the activity was 0.07 U log.These results demonstrate that the nucleus transformation of C.reinhardtii can function as a stable expression platform for the production of the synthetic gene E-760 and it can potentially be used as an antibacterial agent.展开更多
Indigenous fermented foods are known for their nutritional and functional properties but they are often spoiled by pathogenic bacteria that can constitute a food safety problem. “Gappal” is a no-thermal treat food b...Indigenous fermented foods are known for their nutritional and functional properties but they are often spoiled by pathogenic bacteria that can constitute a food safety problem. “Gappal” is a no-thermal treat food based on millet dough and milk and its production conditions can constitute a food safety problem. The aim of this study was to screen and identify LAB producing Bacteriocin-like inhibitory substances using a matrix similar to “Gappal”. The detection of potential BLIS was first performed using overlaid method after enrichment of samples in whey and millet dough. The isolates demonstrating inhibiting area were preselected, purified and tested for the presence of antibacterial properties using their neutralized cell-free culture supernatant and subsequently treated with catalase in combination with protease, pepsin or trypsin. The antimicrobial effect of two isolates (Gbf48 and Gbf50) after growth on MRS broth over 12 h at 30?C were active against E. faecalis ATCC 19433, M. luteus ATCC 49732, S. aureus ATCC 2523, L. monocytogenes, B. megaterium, B. sphaericus and B. cereus with an activity of 2560 AU/mL. The 16S RNA gene sequencing identification indicated that these isolates are Pediococcus acidilactici. Gbf 48 and Gbf 50 could be used to improve preservative factors for a controlled fermentation of non thermal treatment fermented food for their potential of acidification adds to BLIS production.展开更多
This study aimed to determine the antibiotic and bacteriocin sensitivity of Listeria monocytogenes strains isolated from animal derived foods. With disc diffusion assay, all fourteen L. monocytogenes strains were susc...This study aimed to determine the antibiotic and bacteriocin sensitivity of Listeria monocytogenes strains isolated from animal derived foods. With disc diffusion assay, all fourteen L. monocytogenes strains were suscepti-ble to the antibiotics, including penicillin G, vancomycin, tetracycline, chloramphenicol, rifampicin, erythromycin, gentamicin and trime- thoprim. However, the percentages of fosfomycin and streptomycin resistances were 92.9% and 7.1%, respectively. Multiple resistances were not observed among the tested strains. The results of well diffusion assays showed that all strains were inhibited by the cell-free supernatant of a bacteriocin-producing strain, Pediococcus acidilactici 13, with the inhibition zones ranging from 16.00 to 24.50 mm. These results provide useful information on antibiotic resistance of L. monocytogenes strains isolated from foods, and can potentially be used to develop bacteriocin-based interventions to guard against the hazards associated with L. monocytogenes in ready-to-eat meat and poultry products.展开更多
Many lactic acid bacteria (LAB) were isolated from “Shermout”, a popular Sudanese fermented beef product intended for long storage. An isolate that demonstrated significant antibacterial activity was identified as L...Many lactic acid bacteria (LAB) were isolated from “Shermout”, a popular Sudanese fermented beef product intended for long storage. An isolate that demonstrated significant antibacterial activity was identified as Lactobacillus plantarum PM4 based on phenotypic, physiological and biochemical characteristics and carbohydrate utilization patterns. The inhibitory activity of the partially purified bacteriocin was completely arrested by the proteolytic enzymes proteinase-k and pepsin but not by α-amylase, asserting its proteinaceous nature. The activity was not due to H2O2 as similar inhibition was obtained by cell-free supernatant (CFS) produced under anaerobic conditions. The bacteriocin showed a molecular weight in the range of 3 - 5 kDa and had a bactericidal mode of action. No significant reduction in activity was observed on heating to 60℃ for 60 min, but activity was lost on heating to 100℃ or autoclaving. Highest inhibitory activity was at pH 5.5 and there was appreciable reduction in activity at pH 3, 7 or 9. There was no drop in activity at –80℃ or –20℃ up to four weeks of storage. However, at 4℃ and 35℃, a gradual decline in activity was observed. Lb. plantarum PM4 exhibited bactericidal activity against Staphylococcus aureus, Bacillus subtilis, Enterococcus faecalis, Escherichia coli ATCC25922, Klebsiella pneumoniae and Proteus vulgaris. Bacteriocin production generally coincided with the phase of maximum growth and the best combination for maximum production of inhibitory activity was at pH 5.5 for 48 h whether incubated at 25℃, 30℃ or 37℃. Lb. plantarum PM4 showed promise as a starter culture in the fermentation of preserved meat products.展开更多
In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY-3, which produced a bacteriocin-like substance(BLS), was characterized to be a strain of Bacillus...In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY-3, which produced a bacteriocin-like substance(BLS), was characterized to be a strain of Bacillus subtillis by biochemical profiling and 16 S r DNA sequence analysis. It is the first time to report that Bacillus subtilis from Jiaozhou Bay sediments could produce a BLS. The BLS of B. subtillis SLYY-3 exhibited strong inhibitory activity against gram-positive bacteria(including Staphylococcus aureus and B. subtillis) and some fungi(including Penicillium glaucum, Aspergillus niger and Aspergillus flavus). The antimicrobial activity was detected from culture in the exponential growth phase and reached its maximum when culture entered into stationary growth phase. It was thermo-tolerant even when being kept at 100℃ for 60 min without losing any activity and stable over a wide p H range from 1.0 to 12.0 while being inactivated by proteolytic enzyme and trypsin, indicating the proteinaceous nature of the BLS. The BLS was purified by precipitation with hydrochloric acid(HCl) and gel filteration(Sephadex G-100). SDS-PAGE analysis of the extracellular peptides of SLYY-3 revealed a bacteriocin-like protein with a molecular mass of 66 k Da. Altogether, these characteristics indicate the potential of the BLS for food industry as a protection against pathogenic and spoilage microorganisms.展开更多
Direct-fed microbials(DFM), generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other ha...Direct-fed microbials(DFM), generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other hand, methanogenesis in rumen, which accounts for a significant loss of ruminant energy and increased greenhouse gas in environment, is of great concern, therefore, use of DFM for improving productivity without compromising the animal health and ecological sustainability is encouraged. The present study was conducted to investigate the methane reducing potential of bacteriocinogenic strain Pediococcus pentosaceus-34. Since, the culture showed no hemolysis on blood agar and DNase activity, hence, it was considered to be avirulent in nature, a prerequisite for any DFM. The culture also showed tolerance to pH 5.0 for 24 h with 0.5% organic acid mixture, whereas when given a shock for 2 h at different p H and organic acids concentrations, it showed growth at pH 3.0 and 4.0 with 0.1 and 1.0% organic acids, respectively, as having good animal probiotics attributes. The total gas production was significantly(P<0.05) higher in live pedicoccal culture(LPC) and dead pedicoccal culture(DPC) both with wheat straw, when compared to the control. In sugarcane bagasse, gas production was significantly lower(P<0.05) with LPC compared to the control and DPC both. Methane was reduced by the inclusion of LPC in sugarcane bagasse(0.07 mL CH4 mg–1 dry matter digestibility) with no effect on other rumen fermentation parameters. However, with wheat straw and LPC total gas, in vitro dry matter digestibility, total volatile fatty acids increased significantly but no reduction in methane production was observed in comparison to the control. Therefore, further research is warranted in this direction, if the bacteriocinogenic strains can be used as DFM for ruminants to improve the ruminant productivity.展开更多
In this paper,four limit cycles are constructed for a concrete 3D model of rock-scissorpaper(RSP)game between bacteriocin producing bacteria.This gives not only an affirmative answer to the conjecture of the existence...In this paper,four limit cycles are constructed for a concrete 3D model of rock-scissorpaper(RSP)game between bacteriocin producing bacteria.This gives not only an affirmative answer to the conjecture of the existence of three limit cycles raised by Zhang and Yan(2017),but also extends to an construction of four limit cycles.展开更多
基金supported by Sathyabama Institute of Science and Technology,Chennai,Tamil Nadu,India and Indian Council of Medical Research(ICMR),New Delhi,India(Ref.No:5/8/5/19/2014-ECD-I)
文摘Tuberculosis(TB) is a communicable disease caused by Mycobacterium tuberculosis(M. tuberculosis). WHO estimated that 10.4 million new(incident) TB cases worldwide in year 2016. The increased prevalence of drug resistant strains and side effects associated with the current anti-tubercular drugs make the treatment options more complicated. Hence, there are necessities to identify new drug candidates to fight against various sub-populations of M. tuberculosis with less or no toxicity/side effects and shorter treatment duration. Bacteriocins produced by lactic acid bacteria(LAB) attract attention of researchers because of its "Generally recognized as safe" status. LAB and its bacteriocins possess an effective antimicrobial activity against various bacteria and fungi. Interestingly bacteriocins such as nisin and lacticin 3147 have shown antimycobacterial activity in vitro. As probiotics, LAB plays a vital role in promoting various health benefits including ability to modulate immune response against various infectious diseases. LAB and its metabolic products activate immune system and thereby limiting the M. tuberculosis pathogenesis. The protein and peptide engineering techniques paved the ways to obtain hybrid bacteriocin derivatives from the known peptide sequence of existing bacteriocin. In this review, we focus on the antimycobacterial property and immunomodulatory role of LAB and its metabolic products. Techniques for large scale synthesis of potential bacteriocin with multifunctional activity and enhanced stability are also discussed.
基金Managemtent of Vivekananda Educational Inslitulions Tiruchengode, India for their support throughout the project
文摘Objective:To screen the bacteriocinogenic isolate from buffalo milk and to characterize it on physical,chemical and biological aspects for the application in biopreservation.Methods:Bacillus cereus(B.cereus)was isolated and assessed for its baceteriocinogenic activity.Bacteriocin was produced and purified by ammonium sulphate precipitation,dialysis and gel filtration chromatography.Purified bacteriocin was used to clieck its antimicrobial activity against food borne bacteria.Effect and stability of bacteripcin was determined with the respect to temperature,pH,enzymes,organic solvents and chemicals.Bacteriocin was also subjected to SDS PAGE analysis to determine its molecular weight.In addition,functional groups exist in the bacteriocin was determined by FTIR analysis.Results:B.cereus was identified by 16S rRNA sequence analysis.Bacteriocin showed increased activity against all the bacteria used and its activity unit was found to be 51,200 AU/mL.It was stable to high temperature(100℃)and wide range of pH(3-10),sensitive to proteolytic enzymes and resistant to nonprotcolytic enzymes.It was low molecular weight(3.5-6 kDa)protein and FTIR study revealed the presence of amide group and NH stretching,Conclusions:Bacteriocin produced in this study possesses the highest antimicrobial activity against both gram positive and gram negative bacteria thereby it has immense application as biopreservative agent.FTIR proved its peptide nature.
文摘Biopreservation systems in foods are of increasing interest for industry and consumers. Bacteriocin producing Lactobacillus spp. is considered Generally Recognize as Safe (GRAS), useful to control the fast development of pathogens and spoiling microbes in food and feed. Lactobacillus spp. was isolated from traditional winter fermented vegetable cucumber & carrot by the use of selective media. Especially De Man, Rogosa and Sharpe (MRS) Agar media were used to isolate the Lactobacillus species. Morphologically identified by gram staining & colony morphology. Biochemically recognized by catalase, oxidase, MRVP & carbohydrate fermentation test. Antimicrobial activity of Lactobacillus spp. was confirmed by Well Diffusion Method. Molecular characterization of bacteriocin protein and molecular weight determined by SDS PAGE method. The isolate was found to be facultative anaerobic, Gram positive, and catalase negative. The result of antimicrobial activity measured by the Arbitrary Unit (AU/ml) of zone of the inhibition. Six isolates found from the sample but most activities exhibited isolate 4 against Bacillus megaterium (55 mm) zone of diameter. The molecular weight of the washed bacteriocin was calculated to be about 40 kDa (Isolate 1) and 15 kDa & 30 kDa (Isolate 5). Bacteriocin protein reduces chemical preservatives and uses in future as biopreservative in food industry.
文摘Lactobacilli belong to the group of lactic acid bacteria (LAB), that have several distinguished abilities such as production of lactic acid, enzymes such as β-Galactosidase and natural antimicrobial substances called bacteriocins. Bacteriocin is a biopreservative agent potential of suppressing growth of some contaminant bacteria in food industry but its commercial availability is limited and costly. The study aimed to select isolates of Lactobacillus spp. potential for producing bacteriocins to suppress the growth of Escherichia coli ATCC 25922 and Bacillus subtilis NCIB3610, and to optimize the process of bacteriocin production. Results obtained in this study showed that L. acidophilus isolate CH1 was selected as the best candidate for bacteriocin among the four isolates that tested. The largest amounts of the bacteriocins were synthesized only in MRS medium was supplemented with K2HPO4 (1.0%), Tween 80 (1%), Beef extract (1%), glucose, cyctein and peptone extract (1%). The optimization of culture conditions for bacteriocin production areas showed that corn steep liquor medium was the best medium for all isolates against Bacillus subtilis while no effect was observed on Escherichia coli ATCC 25922 except when used MRS medium. The optimum conditions for bacteriocin production were pH 6.0, temperature 34?C with 4% Phenyl acetamide showing the greatest growth inhibition areas.
文摘A bacteriocin producing strain of Lactococcus lactis subsp. lactis LABW4 was isolated from naturally fermented milk product which exhibited strong antibacterial activity against Listeria monocytogenes MTCC657, a food spoilage psychrophilic organism. Both cell free and heat killed supernatants of LABW4 were effective to produce zones of inhibition against L. monocytogenes in vitro. The antibacterial metabolite(s) of LABW4 showed strong cidal effect on the growth of L. monocytogenes. Meat samples, mixed with heat killed supernatant of LABW4 when inoculated with Listeria, remain fresh up to 25 days in refrigerated condition whereas spoilage started immediately after 24 hours of inoculation for control sets. Enhancement of Lactate dehydrogenase of L. monocytogenes upon treatment with LABW4 cell free supernatant suggested its lytic mode of action. Cell lysis or degradations were also supported by scanning electron micrograph of treated cells.
文摘A collection of lactic acid bacteria isolates from fish viscera was studied and investigated regarding to their functional properties and safety aspects. From these, three isolates GM1, GM2 and GM3 were identified as Enterococcus feacium species using molecular methods. Partial Amplified rDNA Restriction Analysis (partial ARDRA) with restriction enzyme HaeIII separated these isolates into distinctive group which suggest genotypic variability within enterococci strains isolated from fish viscera. The three strains GM1, GM2 and GM3 exhibited antimicrobial activity. Indeed strains have been shown to produce bacteriocins with inhibitory effect against food spoilage bacteria and pathogenic fish including Carnobacterium maltaromaticum. The molecular mass of bacteriocin, as calculated by tricine-SDS-PAGE, was found to be 4.5 kDa. All isolates were tested positive upon PCR amplification of enterocin A structural gene. Investigations of antibiotic resistance show that the isolates were mostly sensitive to several antibiotics (ampicillin, penicillin, tetracycline, gentamycine) and resistance to rifampicin. All isolates grow in esculin azide agar as a selective medium for enumeration of probiotic enterococci. This study suggests that our strains can be employed as probiotic or to improve the safety of food products.
文摘Two lactic acid bacteria strains (At1BEAE22 and At344E21) isolated during tchapalo production were identified on the basis of phenotypic analyses. Bacteriocins produced by these strains were tested for their antimicrobial activities using well diffusion agar method. Heat resistance, pH sensitivity and enzyme treatments were also analyzed. Results showed that both lactic acid bacteria strains were identified as Pediococcus acidilactici. Their bacteriocins inhibited growth of Lactobacillus delbrueckii F/31, Listeria innocua ATCC 33090, Enterococcus faecalis, Enterococcus faecalis ATCC 29212, Streptococcus sp, Enterococcus faecalis CIP 105042 and Enterococcus faecium ATCC 51558. These bacteriocins were heat stable at 60°C for 30 min for all indicator bacteria. However, they remained active only against Lactobacillus delbrueckii and Listeria innocua at 121°C for 60 min. Moreover, they were active in a wide range of pH (3 to 9) with a maximum activity observed at pH 5 and 6 on all indicator bacteria. But, bacteriocin from Pediococcus acidilactici At34E21 was more stable at acidic pH than basic one. The fact that the bacteriocin was inactivated by proteinase K and α-chymotrypsin indicated its proteinaceous nature, a general characteristics of bacteriocins.
文摘Bacteriocins are a large group of chromosome or plasmid-encoded and ribosomally synthesized low-molecular-weight (2 to 6 kDa) antimicrobial and amphiphilous peptides produced by Gr+ or Gr- bacteria [1]. Their low toxicity as well as the absence of allergenicity and reactogenicity is confirmed by testing selected bacteriocins [2] [3]. Bacteriocins can be widely used as preservatives and antibiotic alternatives in medicine. Nisin, a Streptococcus lactis-derived bacteriocin, has been in practice in food industry for a long time. A relevant product contains about 2.5% of nisin. For medical use (e.g., when injected into the blood stream), highly purified drugs are required. However, the yield of bacteriocins accounts for no more than a few percents from the total activity in the culture liquid. In this paper, we propose methods (by example of two B. subtilis strains), allowing to increase the yield up to ~80%. It is believed that other bacteriocins may be purified by these methods and with the same yield.
基金supported partly by TWAS Research Grant for Basic Science(10-088 RG/BIO/AS_G-UNESCO FR:3240246003 and 14-094 RG/BIO/AS_G-UNESCO FR:324028606)to A.M.a New Zealand Dental Association Research Foundation grant to N.H
文摘Objective: To characterize the bacteriocin-like inhibitory-substances(BLIS) activity of Streptococcus macedonicus MBF10-2 [named BLIS(MBF10-2)], a bacteriocinogenic strain isolated from an Indonesian tofu byproduct.Methods: BLIS(MBF10-2) was obtained by culturing the bacterium, and standard deferred antagonism assays were used to demonstrate its activity. The antibacterial testing of fractions collected by filtration using 3–30 kDa cut-off membrane sizes were carried out by performing well diffusion method.Results: The growth of Micrococcus luteus, Streptococcus pyogenes, Lactococcus lactis,Leuconostoc mesenteroides and Weissella confusa were inhibited by BLIS(MBF10-2).Interestingly, BLIS-containing fractions obtained from sequential application on ultrafiltration membranes indicated that this bacterium Streptococcus macedonicus MBF10-2 could produces at least two antimicrobial peptides activities, one of which is likely to be a lantibiotic peptide. Potential synergistic activity against certain Gram-positive(but not Gram-negative) species when partnered with antibiotics(ampicillin, tetracycline or kanamycin) were observed.Conclusion: Combination of some BLIS(MBF10-2) active fractions with antibiotics(ampicillin, tetracycline or kanamycin) could demonstrate synergistic activities against certain Gram-positive species.
基金supported by the National Natural Science Foundation of China Program(32021005)the Natural Science Foundation of Jiangsu Province(BK20200084)+2 种基金Projects of Innovation and Development Pillar Program for Key Industries in Southern Xinjiang of Xinjiang Production and Construction Corps(2018DB002)National First Class Discipline Program of Food Science and Technology(JUFSTR20180102)the Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province。
文摘This study was performed to determine the effects of bacteriocin-producing and non-bacteriocin-producing Pediococcus acidilactici strains on the immune system and intestinal flora of normal mice.Two P.acidilactici strains with antibacterial activity(P.acidilactici CCFM28 and CCFM18)were obtained based on the inhibition-zone assay.The produced components were identified as bacteriocins through protease treatment,pH adjustment and hydrogen peroxide treatment.Bacteriocin-producing and non-bacteriocin-producing P.acidilactici strains(P.acidilactici CCFM28,CCFM18 and NT17-3)caused significant changes in serum immune factors and intestinal flora of normal mice.After 14 days of intervention,the relative abundance of Firmicutes was significantly decreased,but that of Proteobacteria was significantly increased at the phylum level.At the genus level,the administration of three P.acidilactici strains resulted in the downregulation of Blautia and the upregulation of Ruminococcus and Lactobacillus.Furthermore,there were also different regulations on some probiotic strains,such as Bifidobacterium,Coprococcus and Akkermansia,which were closely related to the antibacterial ability of the bacteriocin and the type of strain.The results indicated that the intervention of different P.acidilactici strains could differently change the structure of intestinal flora in normal mice,which provided theoretical guidance for the selective use of bacteriocin-producing strains for health regulation in the future.
文摘The use of antimicrobial peptides(AMPs)synthesized by bacteria(bacteriocins)is an alternative for combating multidrug resistant bacterial strains and their production by recombinant route is a viable option for their mass production.The bacteriocin E-760 isolated from the genus Enterococcus sp.has been shown to possess inhibitory activity against Gram-negative and Gram-positive bacteria.In this study,the expression of a chimeric protein coding for E-760 in the nucleus of C.reinhardtii was evaluated,as well as,its antibacterial activity.The synthetic gene E-760S was inserted into the genome of C.reinhardtii using Agrobacterium tumefaciens.A transgenic line was identified in TAP medium with hygromycin and also by PCR.The increment in the culture medium temperature of the transgenic strain at 35°C for 10 minutes,increased the production level of the recombinant protein from 0.14(Noninduced culture,NIC)to 0.36%(Induced culture,IC)of total soluble proteins(TSP);this was quantified by an ELISA assay.Recombinant E-760 possesses activity against Staphylococcus aureus in 0.34 U log,Streptococcus agalactiae in 0.48 U log,Enterococcus faecium in 0.36 U log,Pseudomonas aeruginosa in 2 U log and for Klebsiella pneumoniae,the activity was 0.07 U log.These results demonstrate that the nucleus transformation of C.reinhardtii can function as a stable expression platform for the production of the synthetic gene E-760 and it can potentially be used as an antibacterial agent.
文摘Indigenous fermented foods are known for their nutritional and functional properties but they are often spoiled by pathogenic bacteria that can constitute a food safety problem. “Gappal” is a no-thermal treat food based on millet dough and milk and its production conditions can constitute a food safety problem. The aim of this study was to screen and identify LAB producing Bacteriocin-like inhibitory substances using a matrix similar to “Gappal”. The detection of potential BLIS was first performed using overlaid method after enrichment of samples in whey and millet dough. The isolates demonstrating inhibiting area were preselected, purified and tested for the presence of antibacterial properties using their neutralized cell-free culture supernatant and subsequently treated with catalase in combination with protease, pepsin or trypsin. The antimicrobial effect of two isolates (Gbf48 and Gbf50) after growth on MRS broth over 12 h at 30?C were active against E. faecalis ATCC 19433, M. luteus ATCC 49732, S. aureus ATCC 2523, L. monocytogenes, B. megaterium, B. sphaericus and B. cereus with an activity of 2560 AU/mL. The 16S RNA gene sequencing identification indicated that these isolates are Pediococcus acidilactici. Gbf 48 and Gbf 50 could be used to improve preservative factors for a controlled fermentation of non thermal treatment fermented food for their potential of acidification adds to BLIS production.
文摘This study aimed to determine the antibiotic and bacteriocin sensitivity of Listeria monocytogenes strains isolated from animal derived foods. With disc diffusion assay, all fourteen L. monocytogenes strains were suscepti-ble to the antibiotics, including penicillin G, vancomycin, tetracycline, chloramphenicol, rifampicin, erythromycin, gentamicin and trime- thoprim. However, the percentages of fosfomycin and streptomycin resistances were 92.9% and 7.1%, respectively. Multiple resistances were not observed among the tested strains. The results of well diffusion assays showed that all strains were inhibited by the cell-free supernatant of a bacteriocin-producing strain, Pediococcus acidilactici 13, with the inhibition zones ranging from 16.00 to 24.50 mm. These results provide useful information on antibiotic resistance of L. monocytogenes strains isolated from foods, and can potentially be used to develop bacteriocin-based interventions to guard against the hazards associated with L. monocytogenes in ready-to-eat meat and poultry products.
文摘Many lactic acid bacteria (LAB) were isolated from “Shermout”, a popular Sudanese fermented beef product intended for long storage. An isolate that demonstrated significant antibacterial activity was identified as Lactobacillus plantarum PM4 based on phenotypic, physiological and biochemical characteristics and carbohydrate utilization patterns. The inhibitory activity of the partially purified bacteriocin was completely arrested by the proteolytic enzymes proteinase-k and pepsin but not by α-amylase, asserting its proteinaceous nature. The activity was not due to H2O2 as similar inhibition was obtained by cell-free supernatant (CFS) produced under anaerobic conditions. The bacteriocin showed a molecular weight in the range of 3 - 5 kDa and had a bactericidal mode of action. No significant reduction in activity was observed on heating to 60℃ for 60 min, but activity was lost on heating to 100℃ or autoclaving. Highest inhibitory activity was at pH 5.5 and there was appreciable reduction in activity at pH 3, 7 or 9. There was no drop in activity at –80℃ or –20℃ up to four weeks of storage. However, at 4℃ and 35℃, a gradual decline in activity was observed. Lb. plantarum PM4 exhibited bactericidal activity against Staphylococcus aureus, Bacillus subtilis, Enterococcus faecalis, Escherichia coli ATCC25922, Klebsiella pneumoniae and Proteus vulgaris. Bacteriocin production generally coincided with the phase of maximum growth and the best combination for maximum production of inhibitory activity was at pH 5.5 for 48 h whether incubated at 25℃, 30℃ or 37℃. Lb. plantarum PM4 showed promise as a starter culture in the fermentation of preserved meat products.
基金supported by the National Science and Technology Support Program (No. 2011BAD14B04)Project of Shandong Province Higher Educational Science and Technology Program (J14LE59)+1 种基金Applied & Basic Research Foundation of Qingdao (No. 12-1-4-3(3)-jch)Science & Technology Project of AQSIQ (No. 2012IK176)
文摘In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY-3, which produced a bacteriocin-like substance(BLS), was characterized to be a strain of Bacillus subtillis by biochemical profiling and 16 S r DNA sequence analysis. It is the first time to report that Bacillus subtilis from Jiaozhou Bay sediments could produce a BLS. The BLS of B. subtillis SLYY-3 exhibited strong inhibitory activity against gram-positive bacteria(including Staphylococcus aureus and B. subtillis) and some fungi(including Penicillium glaucum, Aspergillus niger and Aspergillus flavus). The antimicrobial activity was detected from culture in the exponential growth phase and reached its maximum when culture entered into stationary growth phase. It was thermo-tolerant even when being kept at 100℃ for 60 min without losing any activity and stable over a wide p H range from 1.0 to 12.0 while being inactivated by proteolytic enzyme and trypsin, indicating the proteinaceous nature of the BLS. The BLS was purified by precipitation with hydrochloric acid(HCl) and gel filteration(Sephadex G-100). SDS-PAGE analysis of the extracellular peptides of SLYY-3 revealed a bacteriocin-like protein with a molecular mass of 66 k Da. Altogether, these characteristics indicate the potential of the BLS for food industry as a protection against pathogenic and spoilage microorganisms.
基金a part of a PhD project of Sanjay Kumar that was supported by NDRI (ICAR) fellowshipNational Initiative on Climate Resilient Agriculture, India (NICRA) for providing partial support
文摘Direct-fed microbials(DFM), generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other hand, methanogenesis in rumen, which accounts for a significant loss of ruminant energy and increased greenhouse gas in environment, is of great concern, therefore, use of DFM for improving productivity without compromising the animal health and ecological sustainability is encouraged. The present study was conducted to investigate the methane reducing potential of bacteriocinogenic strain Pediococcus pentosaceus-34. Since, the culture showed no hemolysis on blood agar and DNase activity, hence, it was considered to be avirulent in nature, a prerequisite for any DFM. The culture also showed tolerance to pH 5.0 for 24 h with 0.5% organic acid mixture, whereas when given a shock for 2 h at different p H and organic acids concentrations, it showed growth at pH 3.0 and 4.0 with 0.1 and 1.0% organic acids, respectively, as having good animal probiotics attributes. The total gas production was significantly(P<0.05) higher in live pedicoccal culture(LPC) and dead pedicoccal culture(DPC) both with wheat straw, when compared to the control. In sugarcane bagasse, gas production was significantly lower(P<0.05) with LPC compared to the control and DPC both. Methane was reduced by the inclusion of LPC in sugarcane bagasse(0.07 mL CH4 mg–1 dry matter digestibility) with no effect on other rumen fermentation parameters. However, with wheat straw and LPC total gas, in vitro dry matter digestibility, total volatile fatty acids increased significantly but no reduction in methane production was observed in comparison to the control. Therefore, further research is warranted in this direction, if the bacteriocinogenic strains can be used as DFM for ruminants to improve the ruminant productivity.
基金supported by the Specialized Research Fund for the Doctoral Program of Higher Education of China under Grant No.20115134110001。
文摘In this paper,four limit cycles are constructed for a concrete 3D model of rock-scissorpaper(RSP)game between bacteriocin producing bacteria.This gives not only an affirmative answer to the conjecture of the existence of three limit cycles raised by Zhang and Yan(2017),but also extends to an construction of four limit cycles.
