MAG
-
Upload
gabriel-costa -
Category
Technology
-
view
332 -
download
3
Transcript of MAG
Gabriel Costa
Evaluation of antimicrobial properties
of MAG
09/11/2009
IntroductionAimMethodsResultsProblemsConclusionAcknowledgments
Out
line
Introduction
IntroductionMonoacylglycerols
CARACHTERISTICS• Partial esters of glycerol • Non-ionic surface-active compounds• Biodegradability
APPLICATION• Food, cosmetic, textile, plastic industry, etc.
PREPARATION• Interesterification• Hydrolysis of triacylglycerols• Fatty acid addition to glycidol
IntroductionNisin
CARACHTERISTICS• Polycyclic peptide antibacterial with 34 aminoacids• Rare example of a broad-spectrum bacteriocin (effective against gram positives and gram negatives)
APPLICATION• Food industry as food preservative
PREPARATION• Fermentation using Lactococcus lactis
Aim
Evaluate antimicrobial effect of monoacylglycerol
Lauric acid Palmitic acidAdamantanecarboxylic
acid
Aim
Nisin
Evaluate of antimicrobial effect of monoacylglycerol
Lauric acid Palmitic acidAdamantanecarboxylic
acid
Aim
Nisin
MAG A
Methods
Glycidol acid:glycidol (1:1,8)
1-Adamantanecarboxylic acid +
toluene (8ml per 1 gram)
Lauric acid or
palmitic acid
glass reactor with a temperature-stabilized jacket
Purification and Filtration
96 ⁰C 30 minutes
MethodsPreparation of MAG
Solvent system Non-solvent system
Chromium (III) acetate hydroxide0,5% w/w
96 ⁰C 30 minutes
96 ⁰C 120 minutes
MethodsAntimicrobial assay
Salmonella enterica ssp. Enterica ser. Enteritidis CCM 4420
Micrococcus luteus CCM 732
Gram +
1. Bacillus subtilis2. Staphylococcus aureus3. Bacillus cereus4. Micrococcus luteus5. Enterococcus Faecalis
Gram -
1. Escherichia coli2. Citrobacter freundii3. Serratia marcescens4. Salmonella enterica5. Pseudomonas aeruginosa
MethodsAntimicrobial assay
Salmonella enterica ssp. Enterica ser. Enteritidis CCM 4420
Micrococcus luteus CCM 732
Gram +
1. Bacillus subtilis2. Staphylococcus aureus3. Bacillus cereus4. Micrococcus luteus5. Enterococcus Faecalis
Gram -
1. Escherichia coli2. Citrobacter freundii3. Serratia marcescens4. Salmonella enterica5. Pseudomonas aeruginosa
Incubation in plate readerAbsorbance readings (600nm)
Growth curves
MethodsAntimicrobial assay
200 µl MPB with MAG + 5 µl of bacterial suspension
Every 30 minutes24 hours
Dilution of MAG in MPB(25 - 1500 µl/ml)
Results
ResultsGram +
ResultsGram +
ResultsGram +
ResultsBacillus subtilis
ResultsBacillus subtilis
41%
ResultsBacillus cereus
ResultsPetri dishes
1. MAG A of Iva - 1000 µg/ml2. MAG A of Gabriel - 1000 µg/ml3. Nisin - 500 µg/ml4. MAG A of Iva + Nisin – 100MAG + 300N µg/ml5. MAG A of Gabriel + Nisin - 100MAG + 300N µg/ml6. MAG Lauric - 1000 µg/ml7. MAG A of Gabriel + Nisin - 200MAG + 200N µg/ml
Used to confirm the results
Problems
ProblemsWe always have...
MAG A• Did not work the first time• 10 days to realize and prepare
another one
PLATE READER• Light bulb burned out•Could not test MAG of palmitic acid•Could not test mix (Nisin + MAG) in gram negative bacterias
Conclusions
Conclusions
COMBINATION• Probable synergistic effect between nisin and MAG• Other combinations are possibly effective• Part of nisin could be replaced by MAG to reduce costs
Acknowledgments
Acknowledgments
Eva SvobodováLenka PlechačováOlga HaukováHanka Miklíková
Special acknowledgments