This document discusses the use of antibiofilm enzymes as an emerging technology for food quality and safety. It begins by explaining how biofilms are formed by microorganisms on surfaces. It then discusses how antibiofilm enzymes can be used to remove biofilms from food contact surfaces as a safer alternative to chemicals. The document classifies the main types of enzymes used for biofilm prevention and removal, including oxidative enzymes, polysaccharide-degrading enzymes, antiquorum sensing enzymes, and proteolytic enzymes. It also explores how combining enzymes with chemical or physical treatments can more effectively disrupt and remove biofilms.

1. Antibiofilm
Enzymes
As an emerging technology for
Food quality and Safety
Group 3:
Nguyễn Quỳnh Trang- BA11-092
Nguyễn Vũ Dương - 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

4. INTRODUCTION
01
Biofilm formation

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

7. Antibiofilm
Enzymes
02

8. ❖ Natural biocatalysts
❖ Increase the rate of
chemical reactions
❖ Biochemical
processes in cells

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

12. Oxidative Enzymes
Polysaccharide-Degrading
Enzymes
Antiquorum Sensing
Enzymes
Proteolytic Enzymes
Antibiofilm Enzymes
Classification

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 5.0μg/mL,
reduced the active biofilm biomass by
approximately 40%
❖ Synergistic effects of DNase (5μg/ mL)
and antibiotics significantly reduced
biofilm biomass.

18. Polysaccharide-Degrading
Enzymes
Combined treatment with
alginate lyase (20μg/mL) and
gentamycin (64μg/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

20. Combining
Enzymes with
Chemical/ Physical
Treatment
04

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

22. ultrasound + 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. 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.

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