This document summarizes a study that isolated bacteriocin-producing Lactobacillus species from the fermented food Dhokla. Lactobacillus delbrueckii was found to produce bacteriocin with strong antimicrobial activity against various bacteria like Bacillus licheniformis and Escherichia coli. The bacteriocin was extracted from the fermented culture and shown to inhibit food spoilage and pathogenic bacteria. This suggests bacteriocin could potentially be used as a natural biopreservative to control harmful bacteria and extend the shelf life of foods.
Optimizing The Bacteriocin Production In Strain Of Lactobacillus pentosus Iso...Innspub Net
MRS agar was used for the isolation of Lactobacillus strain. This strain was recognized through biochemical tests and 16S rRNA ribotyping, as Lactobacillus pentosus. Its antibacterial effects were detected by utilizing cell free supernatant (CFS). Escherichia coli, Bacillus subtilis and Staphylococcus aureus were used as test strains. CFS showed antimicrobial activity against the test strains. CFS was treated with proteinases for the confirmation of loss of antimicrobial activity. Loss of antimicrobial activity on exposure to proteinases indicated the presence of bacteriocin in CFS. CFS was also studied for its antimicrobial effect at different temperatures and pH. Optimum antimicrobial effect was recorded at pH 7 and at temperature 45°C. The current study indicates the antimicrobial activity of strain of L. pentosus against E. coli, B. subtilis and S. aureus.
Optimizing The Bacteriocin Production In Strain Of Lactobacillus pentosus Iso...Innspub Net
MRS agar was used for the isolation of Lactobacillus strain. This strain was recognized through biochemical tests and 16S rRNA ribotyping, as Lactobacillus pentosus. Its antibacterial effects were detected by utilizing cell free supernatant (CFS). Escherichia coli, Bacillus subtilis and Staphylococcus aureus were used as test strains. CFS showed antimicrobial activity against the test strains. CFS was treated with proteinases for the confirmation of loss of antimicrobial activity. Loss of antimicrobial activity on exposure to proteinases indicated the presence of bacteriocin in CFS. CFS was also studied for its antimicrobial effect at different temperatures and pH. Optimum antimicrobial effect was recorded at pH 7 and at temperature 45°C. The current study indicates the antimicrobial activity of strain of L. pentosus against E. coli, B. subtilis and S. aureus.
An overview of lactic acid bacteria - IJBInnspub Net
Lactic acid bacteria (LAB) are renowned for the potential of producing antimicrobial compound and other value added products. Undeniable to concern these probiotic has contributed to the importance of human life. Deserving an attention for its capabilities, this paper will discuss on the general description of lactic acid bacteria, genetics, metabolism and its application to the industries. Get the full articles at: http://www.innspub.net/volume-1-number-3-june-2011/
Scope, roles and responsibilities of microbiologist inAuricle Nissim
useful for everyone interested in pharmaceutical microbiology, helpful for interviews, most asked question for candidates appearing interviews in pharmaceutical ,
Agents of food spoilage; enzymes and chemical agents.
The role of microorganisms in food spoilage and organisms associated with deterioration of foods.
The role of temperature in food spoilage.
Anti-Microbial Activity of probiotic Lactobacilli and Optimization of Bacteri...ijsrd.com
The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.
An overview of lactic acid bacteria - IJBInnspub Net
Lactic acid bacteria (LAB) are renowned for the potential of producing antimicrobial compound and other value added products. Undeniable to concern these probiotic has contributed to the importance of human life. Deserving an attention for its capabilities, this paper will discuss on the general description of lactic acid bacteria, genetics, metabolism and its application to the industries. Get the full articles at: http://www.innspub.net/volume-1-number-3-june-2011/
Scope, roles and responsibilities of microbiologist inAuricle Nissim
useful for everyone interested in pharmaceutical microbiology, helpful for interviews, most asked question for candidates appearing interviews in pharmaceutical ,
Agents of food spoilage; enzymes and chemical agents.
The role of microorganisms in food spoilage and organisms associated with deterioration of foods.
The role of temperature in food spoilage.
Anti-Microbial Activity of probiotic Lactobacilli and Optimization of Bacteri...ijsrd.com
The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.
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Isolation and Characterization of Bacteriocin Producing Lactic Acid Bacteria ...inventionjournals
Bacteriocins are the extracellular proteins produced by the bacteria which inhibit the growth of similar or closely related bacterial strains. Lactic Acid Bacteria (LAB) are predominantly present in the fermented foods produce bacteriocins. In the present study, six strains (three strains each) were isolated from the fermented Bengal gram samples containing with husk and without husk.The organisms isolated were identified as Pediococcussp and Yeast sp.by the biochemical characterization. The bacteriocins produced by these organisms effectively inhibited the growth of E.coli, Klebsiellasp, Staphylococcus aureus, and Pseudomonas aerugenosabut couldn’t inhibit the growth of Proteus vulgaris and Bacillus sp. Strains c2 and c3 showed greater activity at low pH. All the strains showed antibacterial activity against Bacillus sp,E.coli, Klebsiellasp, Staphylococcus aureusand Pseudomonas aerugenosabut Proteus vulgaris showed high resistance when bacteriocin was exposed to temperatures 37°C and 80°C.
Isolation and Characterization of Bacteriocin Producing Lactic Acid Bacteria ...inventionjournals
Bacteriocins are the extracellular proteins produced by the bacteria which inhibit the growth of similar or closely related bacterial strains. Lactic Acid Bacteria (LAB) are predominantly present in the fermented foods produce bacteriocins. In the present study, six strains (three strains each) were isolated from the fermented Bengal gram samples containing with husk and without husk.The organisms isolated were identified as Pediococcussp and Yeast sp.by the biochemical characterization. The bacteriocins produced by these organisms effectively inhibited the growth of E.coli, Klebsiellasp, Staphylococcus aureus, and Pseudomonas aerugenosabut couldn’t inhibit the growth of Proteus vulgaris and Bacillus sp. Strains c2 and c3 showed greater activity at low pH. All the strains showed antibacterial activity against Bacillus sp,E.coli, Klebsiellasp, Staphylococcus aureusand Pseudomonas aerugenosabut Proteus vulgaris showed high resistance when bacteriocin was exposed to temperatures 37°C and 80°C.
Isolation and Characterization of Bacteriocin Producing Lactic Acid Bacteria ...inventionjournals
Bacteriocins are the extracellular proteins produced by the bacteria which inhibit the growth of similar or closely related bacterial strains. Lactic Acid Bacteria (LAB) are predominantly present in the fermented foods produce bacteriocins. In the present study, six strains (three strains each) were isolated from the fermented Bengal gram samples containing with husk and without husk.The organisms isolated were identified as Pediococcussp and Yeast sp.by the biochemical characterization. The bacteriocins produced by these organisms effectively inhibited the growth of E.coli, Klebsiellasp, Staphylococcus aureus, and Pseudomonas aerugenosabut couldn’t inhibit the growth of Proteus vulgaris and Bacillus sp. Strains c2 and c3 showed greater activity at low pH. All the strains showed antibacterial activity against Bacillus sp,E.coli, Klebsiellasp, Staphylococcus aureusand Pseudomonas aerugenosabut Proteus vulgaris showed high resistance when bacteriocin was exposed to temperatures 37°C and 80°C
Preservative potentials of crude bacteriocins produced by Lactobacillus tucce...iosrjce
IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB) covers studies of the chemical processes in living organisms, structure and function of cellular components such as proteins, carbohydrates, lipids, nucleic acids and other biomolecules, chemical properties of important biological molecules, like proteins, in particular the chemistry of enzyme-catalyzed reactions, genetic code (DNA, RNA), protein synthesis, cell membrane transport, and signal transduction. IOSR-JBB is privileged to focus on a wide range of biotechnology as well as high quality articles on genetic engineering, cell and tissue culture technologies, genetics, microbiology, molecular biology, biochemistry, embryology, cell biology, chemical engineering, bioprocess engineering, information technology, biorobotics.
ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER ...American Research Thoughts
Twenty-two lactic acid bacteria (LAB) belonging to Lactobacillus, Lactococcus and
Enterococcus genera were isolated from plants, flowers and other vegetable matrices. The predominant
LAB species were Lactobacillus brevis (57%) followed to Lactobacillus delbrueckii subsp. bulgaricus
(14%) and Lactococcus lactis (14%). Among enterococci E. faecalis (4 /50%), E. faecium (2 /25%), E.
hirae (2 /12,5%) were isolated. The strains were identified previously with biochemical kits and then
confirmed by PCR. Bacteria isolated were tested against nine strains selected among pathogenic and
opportunistic strains to evaluate their capacity to produce BLS (bacteriocin like substance).
Subsequently to demonstrate the horizontal transfer of genes coding for the production of bacteriocin
substances, a conjugation experiment was positively carried out between an Enterococcus faecium
used as donor and a Lactobacillus acidophilus used as recipient.
Antimicrobial activity of Lactobacillus spp. especially (L. planetarium and L. acidophilus) against S. aureus were tested using agar-plug, agar well diffusion methods to select the best isolate that could inhibit the growth of multidrug resistance isolates. Further identification for the presence of bacteriocin was done using ELISA kit. Results showed that Lactobacillus spp isolates were bacteriocin producers with different degrees and that L. planetarium (L7) was the most efficient in bacteriocin production. Therefore, L. planetarium (L7) was selected for purification using 70% saturated ammonium sulfate and gel chromatography. The effect of purified bacteriocin was tested on 16 bacterial isolates using micro-titer plate method and well diffusion method. The results showed the ability of the bacteriocin to inhibit bacteria only at concentrations 1866U/ml (50%), 3732U/ml (100%) with a diameter of inhibition zones ranges between (11-23 mm) respectively. The anti-biofilm activity of purified bacteriocin at concentration 100% was investigated and the results showed that biofilm formation was reduced by 100% in the presence of bacteriocin.
Isolation, Identification of Probiotic Bacteria Present in Milkijtsrd
The aim ofThe aim of this study was to present some data on isolation, growth, and antimicrobial activity, effect of pH, heat, and sensitivity to proteolytic enzymes of lactobacillus as probiotic bacteria. A large amount of probiotic bacteria is present in milk or milk products which are mainly lactic acid bacteria (LAB). Lactic acid bacteria (LAB) including Lactobacillus spp. Isolation, identification of lactic acid bacterial (LAB) was done by Gram' staining and catalase test and further confirmation was based on morphological, cultural, physiological and different biochemical tests. A total four isolates viz. Lactobacillus fermentum, L. casei, L. acidophilus and bifid bacterium longum was identified after different biochemical analysist which were also showed reliable probiotic properties. The antibacterial activity of lactic acid bacteria (LAB) isolated from raw milk against common enteric pathogens. The antagonistic properties of these isolates against Escherichia coli, Staphylococcus aureus, were examined using agar well diffusion method. this study was to present some data on isolation, growth, and antimicrobial activity, effect of pH, heat, and sensitivity to proteolytic enzymes of lactobacillus as probiotic bacteria. A large amount of probiotic bacteria is present in milk or milk products which are mainly lactic acid bacteria (LAB). Lactic acid bacteria (LAB) including Lactobacillus spp. Isolation, identification of lactic acid bacterial (LAB) was done by Gram' staining and catalase test and further confirmation was based on morphological, cultural, physiological and different biochemical tests. A total four isolates viz. Lactobacillus fermentum, L. casei, L. acidophilus and Bifidobacterium longum was identified after different biochemical analysist which were also showed reliable probiotic properties. The antibacterial activity of lactic acid bacteria (LAB) isolated from raw milk against common enteric pathogens. The antagonistic properties of these isolates against Escherichia coli, Staphylococcus aureus, were examined using agar well diffusion method. Shobha Mehra | Vimla Mehra | Dinesh Bhauryal"Isolation, Identification of Probiotic Bacteria Present in Milk" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-5 , August 2018, URL: http://www.ijtsrd.com/papers/ijtsrd16979.pdf http://www.ijtsrd.com/biological-science/biotechnology/16979/isolation-identification-of-probiotic-bacteria-present-in-milk/shobha-mehra
ABSTRACT- Live microorganisms, have beneficial effects on their host’s health, are called as probiotics. There are various possible sources to isolate
these bacteria. In this studyp harmaceutical probiotic sachet is used as isolation source. The purpose of this study is to search the potentiality of
probiotic bacteria and investigate the probiotic properties of isolates.9 different samples of 3 brands of sachet were used for isolation of bacteria.
Isolates were examined according to their probiotic properties. The probiotic characteristics like pH and Bile tolerance, Antagonistic activity and
Antibiotic susceptibility of isolated bacteria Such as Lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium bifidum was done. Bile
Tolerance and pH tolerance was determined with the help of the help of coefficient of growth inhibition if their coefficient of growth inhibition is less
than 0.5 the organism was considered as the pH and Bile tolerance. The Strains of Lactobacillus acidophilus and Lactobacillus rhamnosus and Bifidobacterium
bifidum show best result at the pH Acidic to Neutral (5 to 7) and show a bile tolerance from 1-4 % bile. All the isolated bacteria show
the maximum inhibition against Staphyloccocus aureus and minimum against Salmonella typhi by Lactobacillus Strains but Bifidobacterium show
minimum against Escheria coli. Most isolates show resistance toward antibiotics. From this study it can be concluded that pharmaceutical probiotic
products used in the study were showing satisfactory quality and potential probiotic strain.
Key words- Probiotic, Lactobacillus, Bifidobacterium, Sachet
Isolation studies of Bacteriocin producing Lactic Acid Bacteria from raw goat...Premier Publishers
Probiotics are live microorganisms which when administered in appropriate amount gives health benefit on the host. In the current study raw goat milk obtained from Belapur village (Belapur, Navi Mumbai, Pin-400614, Maharashtra, India) was the source of isolated lactic acid bacteria. Out of 118 screened isolates, only one isolate i.e. RL 76 showed antimicrobial activity upon primary screening. Optimization of the isolate was carried out to check its bacteriocin producing potential. Isolate grew well at 370C and pH 7. Isolate RL 76 tolerated 2% of bile salt and showed no haemolytic activity. Upon morphological and biochemical tests, the isolate RL76 was identified as Lactobacillus casei. The extracted cell free supernatant was partially purified by ammonium sulphate precipitation. Partially purified cell free supernatant showed antibacterial activity against Staphylococcus aureus, Streptococcus fecalis, Klebsiella pneumonia, Pseudomonas aeruginosa, Salmonella typhi and Escherichia coli. The study indicates potential of the isolate as a good probiotic species.
Inhibitory Effects of Lactobacillus acidophilus and Lactobacillus casei Isola...Premier Publishers
The attempt to use probiotic lactic acid bacteria from a popular fermented beverage in a Nigeria (kunun zaki) is a quest to find an ideal cure for Helicobcter pylori-induced gastritis, gastric malignancies and peptic ulcer. The lactic acid bacteria counts of the samples of the beverage were determined and the organisms were identified using standard bacteriological methods and tested against Helicobacter pylori.The results showed the mean Lactic acid bacterial count to be 4.5x 108 cfu/ml while the microbial flora in the beverage were Lactobacillus acidophilus (50%), Lactobacillus casei (20%), Lactobacillus plantarum (10%), Bacillus cereus (10%) and Saccharomyces cerevisiae (10%) with their respective frequencies of occurrences in parentheses. The inhibitory effect of Lactobacillus acidophilus with the mean zone of inhibition of 51.25mm was found to be more effective against Helicobacter pylori strain J99, while Lactobacillus casei with the zone of inhibition of 39.50mm was more effective against strain P12. The synergistic effect of the two lactobacilli combined in equal proportion against both strain J99 and strain P12 with the mean zones of inhibition of 80.00mm and 77.75mm respectively for the H. pylori strains were significantly higher than those of the individual lactobacilli used. It could be concluded from this study that the two Lactobacillus sp from ‘kununzaki’ demonstrated strong inhibitory effects against Helicobacter pylori and further studies are recommended to validate if they could serve as probiotic alternatives to the treatment of H. pylori- induced peptic ulcers.
In vitro controlling of selected human diarrhea causing bacteria by clove ext...Open Access Research Paper
Antibacterial activity of clove extracts (Syzygium aromaticum L.) was proven against five diarrhea causing bacteria. This was further confirmed when compared with commonly used three commercial antibiotics (ciprofloxacin, tetracycline and erythromycin) as a positive control. Significant differences (P<0.0001) were observed in the effect of the antimicrobial agents (clove extracts and antibiotics), and in the sensitivities of the bacterial species (P<0.0001) to the antimicrobial agents. Clove extracts had significant (P<0.001) activity with the acetone extract demonstrating highest activity followed by antibiotics and other extracts against tested bacteria. The zone of inhibition of clove extracts was ranged from 7.33 to 12.00 mm whereas in antibiotics, it was 0.00 to 11.67 mm. Of all the bacteria, Salmonella typhimurium was the most susceptible against all of the extracts as well as concentrations of clove, while low MIC (180 mgml-1) and MBC (680 mgml-1) of the extracts were observed against Shigella dysenteriae. Consequently, clove has a significant antidiarrheal activity and it could be used as an effective antibacterial agent, alternative to the use of antibiotics.
“Microbiological Analysis Of Soy Milk Produced From Soybean”iosrjce
The soy milk was prepared using the soaked soybean seeds then soy milk was treated with Na2CO3
and NaHCO3 for the preservation. By spread plate method and streak plate method two bacteria Escherichia
spp and Streptococcus spp were isolated respectively .These bacteria were subjected to biochemical and
microbiological analysis. In Biochemical analysis IMVIC test was performed. Both the bacteria Escherichia spp
and Streptococcus spp are positive for Methyl red test and citrate utilization whereas both the bacteria are
negative for Voges-Proskauer test and for Indole production test Escherichia spp is positive and Streptococcus
spp is negative. In case of microbiological test Escherichia spp is gram negative and Streptococcus spp is gram
positive. Both these bacteria have catalase production ability
Bioactivity screening of Soil bacteria against human pathogenspharmaindexing
Microorganisms have a profound effect on medical science as they not only infect & cause disease but also produce metabolic products that can cure infections. Soil happens to be a source for a variety of microorganisms. Most of the bacteria, particularly actinomycetes produce biologically active secondary metabolites. Though there are a number of antibiotics available, there is a pressing need for the discovery of new source for antimicrobials against the pathogens due to the development of drug resistance of the pathogenic microorganisms. In addition to, new pathogenic strains are also developing and causing infection to human beings. Bioactive compounds are compounds that are produced by any living organism and are known to exhibit various biological activities both in-vitro & in-vivo. Bioactivity may be antimicrobial, antineoplastic, anticancerous, immunomodulation, antifertility & others. Soil bacteria were isolated by standard technique and by making use of selective media. The isolates were identified and subjected for preliminary screening to look for their ability to produce bioactive materials. A total of 96 strains were isolated from three different soil samples. 14 of them were found to have antibacterial activity against the human pathogens like Staphylococcus aureus, Streptococcus faecalis, E.coli, Klebsiella aerogenes, Proteus vulgaris, Pseudomonas aureginosa and Salmonella typhi by preliminary screening. Further the selected (3) bacteria were grown in the suitable culture media for the production of bioactive metabolites by using rotary shake flask. The active metabolites was isolated by solvent extraction and concentrated by evaporation under reduced pressure. The antimicrobial screening of the active metabolites showed prominent effect against the clinical pathogens under the study.
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Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
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Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
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Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
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Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
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Isolation of Bacteriocin Producing Lactobacillus SPP from fermented food like Dhokla and their antimicrobial assay
1. IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB)
e-ISSN: XXXX-XXXX, p-ISSN: XXXX-XXXX, Volume 1, Issue 3 (Mar. – Apr. 2015), PP 15-17
www.iosrjournals.org
www.iosrjournals.org 15 | Page
Isolation of Bacteriocin Producing Lactobacillus SPP from
fermented food like Dhokla and their antimicrobial assay
Shinde H. S2
, Waranghuse L. R1
and V. A. Mane3
Department of Post-Harvest and Food Biotechnology,
Department of plant Biotechnology
1, 2,3
K. K. Wagh College of Agricultural Biotechnology, Nashik. 422003, India
Abstract: In present study bacteriocin producing Lactobacillus spp was isolated from dhokla and bacteriocin
was produced from Lactobacillus spp by fermentation. Bacteriocin shows strong antimicrobial activity against
Bacillus licheniformis, Streptococcus thermophillus, Streptococcus acidophilus, Escherichia coli and
Zymomonas anaerobia. This study revealed the possibility of using bacteriocin as biopreservative to control
food spoilage and pathogenic bacteria
Keywords: bacteriocin, Dhokla, biopreservative, Lactobacillus spp.
I. Introduction
Dhokla is a traditional fermented food of India and is prepared in almost every
household of India. Origin of Dhokla is Gujarat, India. Dhokla has antimicrobial property as
Lactic acid bacteria (LAB) spp present in Dhokla produces bacteriocin. Bacteriocin also acts
as biopreservative (Saavadogo Aly et al., 2006).
II. Materials and Methods
Test microorganism.
The test microorganisms namely Bacillus licheniformis, Streptococcus thermophillus,
Streptococcus acidophilus, Escherichia coli and Zymomonas anaerobia were procured from
National Chemical Laboratory (NCL), Pune, Maharashtra, India
Isolation of LAB from Dhokla
Dhokla was procured from Local market of Nashik, India. Plate count agar (PCA) and
(Mans Ragosa and Sharpe) MRS broth were used for isolation of LAB (De Man et al., 1960).
The Dhokla sample were homogenized using morter and pestle using distilled water
and after serial dilution pour plated on PCA plates. Inoculated PCA plates were incubated at
37o
C for 72 h. Isolated colonies with typical white colour characteristics were picked from
plated and inoculated in MRS broth.
The cultures of LAB were identified by biochemical, microbiological, physiological
and morphological characteristics. (Neetu Singh et al., 2013). The biochemical tests used
were Indole production, Methyl red, Voges-Proskauer, citrate utilization and production of
Catalase among microbiological test gram staining and motility tests were conducted. (De
Vuyst et al., 2007).
Maintenance of microorganisms
The cultures of LAB were maintained at 4o
C in MRS broth. Test organisms were
maintained at 4o
C on nutrient agar slants. All the bacterial cultures were subcultured at 20
days interval.
Production of Bacteriocin
Isolated LAB cultures were inoculated in 150ml of MRS broth and incubated
anaerobically at 37o
C for 36h. The LAB cells were killed by heating at 80o
C for 10minutes
2. Isolation of Bacterioc in Producing Lactobacillus SPP from fermented food like Dhoklaand ….
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followed bycentrifugation of broth at 8000 rpm for 30minutes. The resultant cell debris was
discarded and cell free supernatant was collected. The pH of collectedsupernatant collected
was then adjusted to 5.0 with 1N NaOH, then extract was concentrated using rotary flash
evaporator and the solution thus obtained is crude bacteriocin. For synergistic activity
bacteriocin was mixed with 1 ml of DMSO and filter sterilized using 0.22µm membrane filter
paper (Vijay Pal et al., 2005)
Agar well diffusion assay
200µl of the 18h old test cultures were inoculated on nutrient agar plates by spread
plate method. 4 wells of diameter 8mm were made in each of the plates. These plates were
filled with 100µl of concentrated bacteriocin and the plates were incubated at 37o
C for
24hours (Schillinger and Lucke, 1989). The inhibition zone was measured in millimeter scale
using antibiotic zone scale (HiMedia, Mumbai)
III. Result and Discussion
Among 17 positive colonies picked from MRS broth 4 were identified as LAB by
biochemical and microbiological tests.
Antimicrobial activity of bacteriocin
Among 4 isolates of LAB, Lactobacillus delbrueckii was selected for the production of
bacteriocin and antimicrobial study against Bacillus licheniformis, Streptococcus
thermophillus, Streptococcus acidophilus, Escherichia coli and Zymomonas anaerobi.Table 3
shows the antibiogram of bacteriocin isolated from Lactobacillus delbrueckii. Standard
antibiotic Erythromycin was used as a positive control and DMSO was used as negative
control. This study proves that fermented food like Dhokla has antimicrobial property due to
bacteriocin produced by Lactobacillus spp. This bacteriocin also acts as a biopreservative by
inhibiting growth of food spoilage causing bacteria. (Boziaris and Adams, 1999).
Table 1 Inhibition zone of antimicrobial activity
Table 2 Biochemical test of bacterial organisms
Sr No Test Name II 10-3
1 Gram staining +
2 Indole test +
3 Methyl red test +
4 Voges - proskauer test +
5 Citrate utilization test -
6 Motility test -
7 Catalase test -
8 Glucose fermentation test -
9 Mannitol test -
(+ Positive; - Negative)
No
Name of microorganism
Zone of inhibition
Bacteriocin in mm(Lactobacillus delbrueckii)
Standard antibiotic in mm
(Erythromycin)
1. Streptococcus thermophillus 21 22
2. Streptococcus acidophillus 19 19
3. Bacillus licheniformis 18 20
4. Zymomonsanerobica 20 19
5. Escherichia coli 24 26
3. Isolation of Bacterioc in Producing Lactobacillus SPP from fermented food like Dhoklaand ….
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Fig 1 inhibition zone of antimicrobial activity
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