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Antibiofilm
Enzymes
As an emerging technology for
Food quality and Safety
Group 3:
Nguyễn Quỳnh Trang- BA11-092
Nguyễn VĆ© DÆ°ÆĄng - BI12-126
Nguyễn Đỗ Minh Nguyệt - BI12-334
Food Industry Concern: Biofilm Formation?
01
02
Table of contents
Introduction
Biofilm formation
Antibiofilm
enzymes
04
Combining
Enzymes
With Chemical/ Physical
Treatment
05
Conclusions
03
Classification of the
main Enzyme
For Biofilm Prevention and
Removal
INTRODUCTION
01
Biofilm formation
Biofilms are polymeric
mixtures (polysaccharides,
lipids, and nucleic acids)
produced by
microorganisms, which
are supported on biotic
and abiotic surfaces.
Scanning electron microscopy (SEM) image of
biofilm formation on polystyrene coupons in A.
hydrophila
Antibiofilm
Enzymes
02
❖ Natural biocatalysts
❖ Increase the rate of
chemical reactions
❖ Biochemical
processes in cells
❖ Biofilm removers from food contact
surfaces.
❖ Replace harmful and inefficient
biocides and reduce the problems
associated with the presence of
biofilms
Classification of the
main Enzyme
for Biofilm
Prevention and
Removal
03
Oxidative Enzymes
Polysaccharide-Degrading
Enzymes
Antiquorum Sensing
Enzymes
Proteolytic Enzymes
Antibiofilm Enzymes
Classification
Antiquorum
Sensing Enzymes
❖ QS is an intercellular communication
mechanism frequently used by several bacterial
species as a result of an increase in the density of
microorganisms.
❖ Based on the production, release, and detection
of small signaling molecules.
❖ Biofilm matrices provide optimal conditions for
QS due to the spatio chemical conditions and
close proximity of cells within them.
Antiquorum Sensing Enzymes
Three components
make up QS system
Classes of signaling
molecules:
acyl homoserine lactones
(AHLs)
autoinducing peptides
(AIPs)
autoinducer-2 (AI-2)
the AI
the gene coding for the AI
synthase protein
the gene coding for the
response regulator protein
❖ All the alternatives used are based on the disruption of some of these
mechanisms.
❖ Four potential cleavage sites in the AHLs are likely cut off after the
catabolic digestion of carbon and nitrogen sources.
Antiquorum Sensing Enzymes
Oxidative
Enzymes
Enzyme-based products can also be targeted to
extracellular DNA (eDNA) found in the biofilm matrix.
The enzymatic degradation of eDNA can prevent,
disperse, or sensitize biofilms to antimicrobials.
Oxidative Enzymes
The effects of deoxyribonuclease (DNase) on the
biofilm of
❖ Grampositive (Staphylococcus aureus and
Streptococcus pyogenes) and
❖ Gram-negative (Acinetobacter baumannii,
Haemophilus influenza, Klebsiella pneumoniae,
E. coli, and P. aeruginosa)
bacteria have been extensively studied.
❖ DNase at a concentration of 5.0ÎŒg/mL,
reduced the active biofilm biomass by
approximately 40%
❖ Synergistic effects of DNase (5ÎŒg/ mL)
and antibiotics significantly reduced
biofilm biomass.
Polysaccharide-Degrading
Enzymes
Combined treatment with
alginate lyase (20ÎŒg/mL) and
gentamycin (64ÎŒg/mL) on a
biofilm formed by two mucoid
P. aeruginosa strains.
Alginate lyase, cellulose, α-
amylase, and lysozyme. The
coadministration of a lyase with
an antibiotic was tested to
inhibit and eradicate biofilms.
Serine proteases effectively reduce Bacillus
biofilms whereas polysaccharides remove
Pseudomonas fluorescens more efficiently
than serine proteases.
Savinase as one of the most effective
enzymatic formulations for eradicating P.
fluorescens biofilms from glass wool.
Proteases are a class of proteolytic enzymes, and
have been shown to hydrolyze protein
macromolecules adhered to the surfaces of food
processing facilities that are difficult to access such
as pipes.
Proteolytic Enzymes
Savinase structure
Combining
Enzymes with
Chemical/ Physical
Treatment
04
❖ Binding enzymes to surface
substrates, dispersing agents, and
chelants:
=> clean, and remove the biofilm
=> Alternative to chemical cleaning
methods
=> More effective than individual methods
❖ Methods:
biocides + enzyme
ultrasound + enzyme
biocides + enzyme
ultrasound + enzyme
Combining Enzymes with
Chemical/ Physical Treatment
Experiments:
● Oulahal: use different enzymes + ultrasound
Remove 75% - 100% biofilm of E. coli and S. aureus
● Akawash: use gentamycin + alginate lyase
Eradicate all of the P. aeruginosa biofilm after 96h
● Oulahal-Lagsir: used proteolytic enzymes +
ultrasound
Remove E. coli biofilms from stainless steel surfaces
● amyloglucosidase (50UmL−1 ) + ultrasound
decrease biofilm appropriate 96%
● α-amylase + buffer with an anionic surfactant
decrease Bacillus biofilm
Advantages
⇒ These methods to increase
disinfectant potency in the food industry
⇒ Cleaner
⇒ effective novel biofilm control
Difficulties
⇒ Determination of effective enzymes
for types of biofilm
⇒ the mode of action is also unknown
Conclusions
biofilm control:
against
pathogenic &
deteriorative
bacteria
Antimicro
bials
Environm
ent
friendly
use at an
industrial scale.
genetically
modified and
highly resistant
bacteria
01 02 03 04
Some references
Tomy J. Gutiérrez, Chapter 19 -
Antibiofilm Enzymes as an
Emerging Technology for Food
Quality and Safety, Editor(s):
Mohammed Kuddus, Enzymes
in Food Biotechnology,
Academic Press, 2019, Pages
321-342, ISBN 9780128132807.
CREDITS: This presentation template was created by
Slidesgo, including icons by Flaticon, and
infographics & images by Freepik
Thank you!
Do you have
any question?

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Antibiofilm Enzymes.pptx

  • 1. Antibiofilm Enzymes As an emerging technology for Food quality and Safety Group 3: Nguyễn Quỳnh Trang- BA11-092 Nguyễn VĆ© DÆ°ÆĄng - BI12-126 Nguyễn Đỗ Minh Nguyệt - BI12-334
  • 2. Food Industry Concern: Biofilm Formation?
  • 3. 01 02 Table of contents Introduction Biofilm formation Antibiofilm enzymes 04 Combining Enzymes With Chemical/ Physical Treatment 05 Conclusions 03 Classification of the main Enzyme For Biofilm Prevention and Removal
  • 5. Biofilms are polymeric mixtures (polysaccharides, lipids, and nucleic acids) produced by microorganisms, which are supported on biotic and abiotic surfaces. Scanning electron microscopy (SEM) image of biofilm formation on polystyrene coupons in A. hydrophila
  • 6.
  • 8. ❖ Natural biocatalysts ❖ Increase the rate of chemical reactions ❖ Biochemical processes in cells
  • 9.
  • 10. ❖ Biofilm removers from food contact surfaces. ❖ Replace harmful and inefficient biocides and reduce the problems associated with the presence of biofilms
  • 11. Classification of the main Enzyme for Biofilm Prevention and Removal 03
  • 13. Antiquorum Sensing Enzymes ❖ QS is an intercellular communication mechanism frequently used by several bacterial species as a result of an increase in the density of microorganisms. ❖ Based on the production, release, and detection of small signaling molecules. ❖ Biofilm matrices provide optimal conditions for QS due to the spatio chemical conditions and close proximity of cells within them.
  • 14. Antiquorum Sensing Enzymes Three components make up QS system Classes of signaling molecules: acyl homoserine lactones (AHLs) autoinducing peptides (AIPs) autoinducer-2 (AI-2) the AI the gene coding for the AI synthase protein the gene coding for the response regulator protein
  • 15. ❖ All the alternatives used are based on the disruption of some of these mechanisms. ❖ Four potential cleavage sites in the AHLs are likely cut off after the catabolic digestion of carbon and nitrogen sources. Antiquorum Sensing Enzymes
  • 16. Oxidative Enzymes Enzyme-based products can also be targeted to extracellular DNA (eDNA) found in the biofilm matrix. The enzymatic degradation of eDNA can prevent, disperse, or sensitize biofilms to antimicrobials.
  • 17. Oxidative Enzymes The effects of deoxyribonuclease (DNase) on the biofilm of ❖ Grampositive (Staphylococcus aureus and Streptococcus pyogenes) and ❖ Gram-negative (Acinetobacter baumannii, Haemophilus influenza, Klebsiella pneumoniae, E. coli, and P. aeruginosa) bacteria have been extensively studied. ❖ DNase at a concentration of 5.0ÎŒg/mL, reduced the active biofilm biomass by approximately 40% ❖ Synergistic effects of DNase (5ÎŒg/ mL) and antibiotics significantly reduced biofilm biomass.
  • 18. Polysaccharide-Degrading Enzymes Combined treatment with alginate lyase (20ÎŒg/mL) and gentamycin (64ÎŒg/mL) on a biofilm formed by two mucoid P. aeruginosa strains. Alginate lyase, cellulose, α- amylase, and lysozyme. The coadministration of a lyase with an antibiotic was tested to inhibit and eradicate biofilms.
  • 19. Serine proteases effectively reduce Bacillus biofilms whereas polysaccharides remove Pseudomonas fluorescens more efficiently than serine proteases. Savinase as one of the most effective enzymatic formulations for eradicating P. fluorescens biofilms from glass wool. Proteases are a class of proteolytic enzymes, and have been shown to hydrolyze protein macromolecules adhered to the surfaces of food processing facilities that are difficult to access such as pipes. Proteolytic Enzymes Savinase structure
  • 21. ❖ Binding enzymes to surface substrates, dispersing agents, and chelants: => clean, and remove the biofilm => Alternative to chemical cleaning methods => More effective than individual methods ❖ Methods: biocides + enzyme ultrasound + enzyme biocides + enzyme
  • 23. Combining Enzymes with Chemical/ Physical Treatment Experiments: ● Oulahal: use different enzymes + ultrasound Remove 75% - 100% biofilm of E. coli and S. aureus ● Akawash: use gentamycin + alginate lyase Eradicate all of the P. aeruginosa biofilm after 96h ● Oulahal-Lagsir: used proteolytic enzymes + ultrasound Remove E. coli biofilms from stainless steel surfaces ● amyloglucosidase (50UmL−1 ) + ultrasound decrease biofilm appropriate 96% ● α-amylase + buffer with an anionic surfactant decrease Bacillus biofilm Advantages ⇒ These methods to increase disinfectant potency in the food industry ⇒ Cleaner ⇒ effective novel biofilm control Difficulties ⇒ Determination of effective enzymes for types of biofilm ⇒ the mode of action is also unknown
  • 24. Conclusions biofilm control: against pathogenic & deteriorative bacteria Antimicro bials Environm ent friendly use at an industrial scale. genetically modified and highly resistant bacteria 01 02 03 04
  • 25. Some references Tomy J. GutiĂ©rrez, Chapter 19 - Antibiofilm Enzymes as an Emerging Technology for Food Quality and Safety, Editor(s): Mohammed Kuddus, Enzymes in Food Biotechnology, Academic Press, 2019, Pages 321-342, ISBN 9780128132807.
  • 26. CREDITS: This presentation template was created by Slidesgo, including icons by Flaticon, and infographics & images by Freepik Thank you! Do you have any question?