Biofilms consist of complex three-dimensional structures produced by fungi and bacteria at interfaces and are considered a severe hazard to human health. The biofilm formed on the surface of medical instruments leads to a major threat of dispersing microorganisms within the host and causing infection through the release of both single and clustered cells. The removal of biofilms is very challenging because of their resistance to antimicrobial therapies and high tolerance toward conventional antimicrobial agents. Therefore, it is necessary to treat biofilms more effectively and also important to understand the mechanism for biofilm formation. Addressing this key issue, this review begins with an outline of the process of biofilm formation and the associated therapeutic strategies, emphasizing the role of lysine oxidase in developing innovative antibiofilm materials for inhibiting and removing biofilms. Amino acid oxidases such as lysine oxidase and escapin are highlighted for their ability to produce hydrogen peroxide (H2O2), which exhibits antimicrobial properties against both planktonic bacteria and biofilm. It was demonstrated that the combined use of escapin intermediate products (EIP) and H2O2 can prevent biofilm formation and disrupt established biofilms at micromolar concentrations by using an organism, Pseudomonas aeruginosa, as an experimental model. In addition was also observed the antifungal activity of lysine and the effects of poly-L-lysine (pLK) on bacterial biofilms. In Marinomonas mediterranea, the lysine oxidase AlpP homologue (LodA) mediates bacterial growth inhibition, DNA strand breakage, lipid peroxidation, and cell death through the accumulation of reactive species such as H2O2. Lysine alone lacked fungicidal activity, but it amplified the action of amphotericin B against C.andida albicans by inhibiting biofilm and hypha formation. Furthermore, pLK showed antimicrobial properties due to its cationic charges, effective against pathogens, including P. aeruginosa and Staphylococcus aureus. The review also includes the potential of synthetic mimics of antimicrobial peptides over natural peptides, which provide better stability and cost-effectiveness for treating biofilms associated with both ventilator-associated pneumonia and cystic fibrosis.
RatnerS. L-Amino acid oxidases (Mammalian Tissues and snake venom), in: ColowickS.P, KaplanN.O. (Eds.) Methods in Enzymology Vol. 2. Academic Press, NY, USA, 1955, pp. 204–211; doi: 10.1016/S0076-6879(55)02188-5
2.
GómezD, Lucas-ElíoP, Sanchez-AmatA, SolanoF. A novel type of lysine oxidase: L-lysine-ε-oxidase. Biochim Biophys Acta, 2006; 1764(10):1577–1585; doi: 10.1016/j.bbapap.2006.08.014
3.
Campillo-BrocalJC, Lucas-ElíoP, Sanchez-AmatA. Distribution in different organisms of amino acid oxidases with FAD or a quinone as cofactor and their role as antimicrobial proteins in marine bacteria. Mar Drugs, 2015; 13(12):7403–7418; doi: 10.3390/md13127073
4.
KusakabeH, KodamaK, KuninakaA, et al.A new antitumor enzyme, L-lysine alpha-oxidase from Trichoderma viride. Purification and enzymological properties. J Biol Chem, 1980; 255(3):976–981; doi: 10.1016/S0021-9258(19)86128-8
5.
PowellLC, PritchardMF, FergusonEL, et al.Targeted disruption of the extracellular polymeric network of Pseudomonas aeruginosa biofilms by alginate oligosaccharides. NPJ Biofilms Microbiomes, 2018; 4(1):13; doi: 10.1038/s41522-018-0056-3
6.
TremblayYD, LévesqueC, SegersRP, JacquesM. Method to grow Actinobacilluspleuropneumoniae biofilm on a biotic surface. BMC Vet Res, 2013; 9:213–218; doi: 10.1186/1746-6148-9-213
7.
StoodleyP, SauerK, DaviesDG, CostertonJW. Biofilms as complex differentiated communities. Annu Rev Microbiol, 2002; 56(1):187–209; doi: 10.1146/annurev.micro.56.012302.160705
PalmerJ, FlintS, BrooksJ. Bacterial cell attachment, the beginning of a biofilm. J Ind Microbiol Biotechnol, 2007; 34(9):577–588; doi: 10.1007/s10295-007-0234-4
MangwaniN, DashHR, ChauhanA, DasS. Bacterial quorum sensing: Functional features and potential applications in biotechnology. J Mol Microbiol Biotechnol, 2012; 22(4):215–227; doi: 10.1159/000341847
12.
LindsayD, Von HolyA. Bacterial biofilms within the clinical setting: What healthcare professionals should know. J Hosp Infect, 2006; 64(4):313–325; doi: 10.1016/j.jhin.2006.06.028
13.
LiuX, YaoH, ZhaoX, GeC. Biofilm formation and control of foodborne pathogenic bacteria. Molecules, 2023; 28(6):2432; doi: 10.3390/molecules28062432
PercivalSL, SulemanL, VuottoC, DonelliG. Healthcare-associated infections, medical devices and biofilms: Risk, tolerance and control. J Med Microbiol, 2015; 64(Pt 4):323–334; doi: 10.1099/jmm.0.000032
16.
DerbyCD, GilbertES, TaiPC. Molecules and mechanisms underlying the antimicrobial activity of escapin, an l-amino acid oxidase from the ink of sea hares. Biol Bull, 2018; 235(1):52–61; doi: 10.1086/699175
17.
JimboM, NakanishiF, SakaiR, MuramotoK, KamiyaH. Characterization of Lamino acid oxidase and antimicrobial activity of aplysianin A, a sea hare-derived antitumor-antimicrobial protein. Fisheries Sci, 2003; 69(6):1240–1246; doi: 10.1111/j.0919-9268.2003.00751.x
18.
JohnsonPM, KicklighterCE, SchmidtM, et al.Packaging of chemicals in the defensive secretory glands of the sea hare Aplysia californica. J Exp Biol, 2006; 209(Pt 1):78–88; doi: 10.1242/jeb.01972
19.
KamioM, KoKC, ZhengS, et al.The chemistry of escapin: Identification and quantification of the components in the complex mixture generated by an L‐amino acid oxidase in the defensive secretion of the sea snail Aplysia californica. Chemistry–A. Chemistry, 2009; 15(7):1597–1603; doi: 10.1002/chem.200801696
20.
SantiagoAJ, AhmedMN, WangSL, et al.Inhibition and dispersal of Pseudomonas aeruginosa biofilms by combination treatment with escapin intermediate products and hydrogen peroxide. Antimicrob Agents Chemother, 2016; 60(9):5554–5562; doi: 10.1128/aac.02984-15
21.
BlackmanLD, QuY, CassP, LocockKE. Approaches for the inhibition and elimination of microbial biofilms using macromolecular agents. Chem Soc Rev, 2021; 50(3):1587–1616; doi: 10.1039/d0cs00986e
22.
Gomes Von BorowskiR, GnoattoSC, MacedoAJ, GilletR. Promising antibiofilm activity of peptidomimetics. Front Microbiol, 2018; 9:2157; doi: 10.3389/fmicb.2018.02157
23.
DuboisAV, MidouxP, GrasD, et al.Poly-L-lysine compacts DNA, kills bacteria, and improves protease inhibition in cystic fibrosis sputum. Am J Respir Crit Care Med, 2013; 188(6):703–709; doi: 10.1164/rccm.201305-0912OC
24.
GuillonA, FouquenetD, MorelloE, et al.Treatment of Pseudomonas aeruginosa biofilm present in endotracheal tubes by poly-L-lysine. Antimicrob Agents Chemother, 2018; 62(11):e00564–e00518; doi: 10.1128/aac.00564-18
25.
Mai-ProchnowA, Lucas-ElioP, EganS, et al.Hydrogen peroxide linked to lysine oxidase activity facilitates biofilm differentiation and dispersal in several gram-negative bacteria. J Bacteriol, 2008; 190(15):5493–5501; doi: 10.1128/jb.00549-08
26.
Andreo-VidalA, Sanchez-AmatA, Campillo-BrocalJC. The Pseudoalteromonasluteoviolacea L-amino acid oxidase with antimicrobial activity is a flavoenzyme. Mar Drugs, 2018; 16(12):499; doi: 10.3390/md16120499
27.
Lucas-ElíoP, GómezD, SolanoF, Sanchez-AmatA. The antimicrobial activity of marinocine, synthesized by Marinomonasmediterranea, is due to hydrogen peroxide generated by its lysine oxidase activity. J Bacteriol, 2006; 188(7):2493–2501; doi: 10.1128/jb.188.7.2493-2501.2006
28.
ChenWM, LinCY, SheuSY. Investigating antimicrobial activity in Rheinheimera sp. due to hydrogen peroxide generated by l-lysine oxidase activity. Enzyme Microb Technol, 2010; 46(6):487–493; doi: 10.1016/j.enzmictec.2010.01.006
29.
Campillo‐BrocalJC, Lucas‐ElioP, Sanchez‐AmatA. Identification in Marinomonasmediterranea of a novel quinoprotein with glycine oxidase activity. Microbiologyopen, 2013; 2(4):684–694; doi: 10.1002/mbo3.107
30.
ZhaoL, JiangJ, ZhuZ, et al.Lysine enhances the effect of amphotericin B against Candida albicans in vitro. Acta Biochimica Et BiophysicaSinica, 2016; 48(2):182–193; doi: 10.1093/abbs/gmv125
31.
BornTL, BlanchardJS. Structure/function studies on enzymes in the diaminopimelate pathway of bacterial cell wall biosynthesis. Curr Opin Chem Biol, 1999; 3(5):607–613; doi: 10.1016/S1367-5931(99)00016-2
32.
DaviesD. Understanding biofilm resistance to antibacterial agents. Nat Rev Drug Discov, 2003; 2(2):114–122; doi: 10.1038/nrd1008
33.
TaylorPK, YeungAT, HancockRE. Antibiotic resistance in Pseudomonas aeruginosa biofilms: Towards the development of novel anti-biofilm therapies. J Biotechnol, 2014; 191:121–130; doi: 10.1016/j.jbiotec.2014.09.003
HyldgaardM, MygindT, VadBS, et al.The antimicrobial mechanism of action of epsilon-poly-l-lysine. Appl Environ Microbiol, 2014; 80(24):7758–7770; doi: 10.1128/AEM.02204-14
Lucas-ElioP, HernandezP, Sanchez-AmatA, SolanoF. Purification and partial characterization of marinocine, a new broad-spectrum antibacterial protein produced by Marinomonasmediterranea. Biochim Biophys Acta, 2005; 1721(1–3):193–203; doi: 10.1016/j.bbagen.2004.11.002
38.
Mai-ProchnowA, EvansF, Dalisay-SaludesD, et al.Biofilm development and cell death in the marine bacterium Pseudoalteromonastunicata. Appl Environ Microbiol, 2004; 70(6):3232–3238; doi: 10.1128/AEM.70.6.3232-3238.2004
39.
BoucherRC. Cystic fibrosis: A disease of vulnerability to airway surface dehydration. Trends Mol Med, 2007; 13(6):231–240; doi: 10.1016/j.molmed.2007.05.001
40.
LivraghiA, RandellSH. Cystic fibrosis and other respiratory diseases of impaired mucus clearance. Toxicol Pathol, 2007; 35(1):116–129; doi: 10.1080/01926230601060025
41.
HauserAR, JainM, Bar-MeirM, McColleySA. Clinical significance of microbial infection and adaptation in cystic fibrosis. Clin Microbiol Rev, 2011; 24(1):29–70; doi: 10.1128/cmr.00036-10
42.
HilpertK, ElliottM, JenssenH, et al.Screening and characterization of surface-tethered cationic peptides for antimicrobial activity. Chem Biol, 2009; 16(1):58–69; doi: 10.1016/j.chembiol.2008.11.006
ChenM, YuQ, SunH. Novel strategies for the prevention and treatment of biofilm related infections. Int J Mol Sci, 2013; 14(9):18488–18501; doi: 10.3390/ijms140918488
45.
KalpanaBJ, AarthyS, PandianSK. Antibiofilm activity of α-amylase from Bacillus subtilis S8-18 against biofilm forming human bacterial pathogens. Appl Biochem Biotechnol, 2012; 167(6):1778–1794; doi: 10.1007/s12010-011-9526-2
46.
WhitchurchCB, Tolker-NielsenT, RagasPC, MattickJS. Extracellular DNA required for bacterial biofilm formation. Science, 2002; 295(5559):1487; doi: 10.1126/science.295.5559.1487
47.
O’TooleGA, KolterR. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol, 1998; 30(2):295–304; doi: 10.1046/j.1365-2958.1998.01062.x
48.
CegelskiL, MarshallGR, EldridgeGR, HultgrenSJ. The biology and future prospects of antivirulence therapies. Nat Rev Microbiol, 2008; 6(1):17–27; doi: 10.1038/nrmicro1818
49.
MatsudaM, AsanoY. Determination of plasma and serum l-lysine using l-lysine ε-oxidase from Marinomonasmediterranea NBRC 103028T. Anal Biochem, 2010; 406(1):19–23; doi: 10.1016/j.ab.2010.06.045
50.
SugawaraA, MatsuiD, KomedaH, AsanoY, IsobeK. Characterization and application of aminoamide-oxidizing enzyme from Aspergillus carbonarius AIU 205. J Biosci Bioeng, 2014; 117(3):263–268; doi: 10.1016/j.jbiosc.2013.08.019
51.
BarbosaJ, BoschE, CortinaJL, RosésM. Improvement of the titrimetric method for the determination of total basicity and available lysine residues in proteinaceous samples in anhydrous acetic acid. Anal Chim Acta, 1992; 256(1):177–181; doi: 10.1016/0003-2670(92)85342-4
52.
WangJ, ZhangP, LiCM, LiYF, HuangCZ. A highly selective and colorimetric assay of lysine by molecular-driven gold nanorods assembly. Biosens Bioelectron, 2012; 34(1):197–201; doi: 10.1016/j.bios.2012.02.001
53.
KimYH, SathiyanarayananG, KimHJ, et al.A liquid-based colorimetric assay of lysine decarboxylase and its application to enzymatic assay. J Microbiol Biotechnol, 2015; 25(12):2110–2115; doi: 10.4014/jmb.1505.05063
54.
PalamakumburaAH, TrackmanPC. A fluorometric assay for detection of lysyl oxidase enzyme activity in biological samples. Anal Biochem, 2002; 300(2):245–251; doi: 10.1006/abio.2001.5464
55.
SinghS, GogoiBK, BezbaruahRL. Optimization of medium and cultivation conditions for L-amino acid oxidase production by Aspergillus fumigatus. Can J Microbiol, 2009; 55(9):1096–1102; doi: 10.1139/W09-068
56.
DandekarT, SnelB, HuynenM, BorkP. Conservation of gene order: A fingerprint of proteins that physically interact. Trends Biochem Sci, 1998; 23(9):324–328; doi: 10.1016/S0968-0004(98)01274-2
57.
GómezD, Lucas‐ElíoP, SolanoF, Sanchez‐AmatA. Both genes in the MarinomonasmediterranealodAB operon are required for the expression of the antimicrobial protein lysine oxidase. Mol Microbiol, 2010; 75(2):462–473; doi: 10.1111/j.1365-2958.2009.07000.x
58.
Chacón-VerdúMD, Campillo-BrocalJC, Lucas-ElíoP, DavidsonVL, Sánchez-AmatA. Characterization of recombinant biosynthetic precursors of the cysteine tryptophylquinonecofactors of l-lysine-epsilon-oxidase and glycine oxidase from Marinomonasmediterranea. Biochim Biophys Acta, 2015; 1854(9):1123–1131; doi: 10.1016/j.bbapap.2014.12.018
59.
Campillo-BrocalJC, Chacón-VerdúMD, Lucas-ElíoP, Sánchez-AmatA. Distribution in microbial genomes of genes similar to lodA and goxA which encode a novel family of quinoproteins with amino acid oxidase activity. BMC Genomics, 2015; 16(1):231–213; doi: 10.1186/s12864-015-1455-y
60.
BoullandML, MarquetJ, Molinier-FrenkelV, et al.Human IL4I1 is a secreted L-phenylalanine oxidase expressed by mature dendritic cells that inhibits T-lymphocyte proliferation. Blood, The Journal of the American Society of Hematology, 2007; 110(1):220–227; doi: 10.1182/blood-2006-07-036210
61.
SunY, NonobeE, KobayashiY, et al.Characterization and expression of L-amino acid oxidase of mouse milk. J Biol Chem, 2002; 277(21):19080–19086; doi: 10.1074/jbc.M200936200
62.
NishiyaY, ImanakaT. Purification and characterization of a novel glycine oxidase from Bacillus subtilis. FEBS Lett, 1998; 438(3):263–266; doi: 10.1016/S0014-5793(98)01313-1
63.
JobV, MarconeGL, PiloneMS, PollegioniL. Glycine oxidase from Bacillus subtilis: Characterization of a new flavoprotein. J Biol Chem, 2002; 277(9):6985–6993; doi: 10.1074/jbc.M111095200
64.
SettembreEC, DorresteinPC, ParkJH, et al.Structural and mechanistic studies on ThiO, a glycine oxidase essential for thiamin biosynthesis in Bacillus subtilis. Biochemistry, 2003; 42(10):2971–2981; doi: 10.1021/bi026916v
65.
Martínez‐MartínezI, Navarro‐FernándezJ, García‐CarmonaF, TakamiH, Sánchez‐FerrerÁ. Characterization and structural modeling of a novel thermostable glycine oxidase from Geobacilluskaustophilus HTA426. Proteins, 2008; 70(4):1429–1441; doi: 10.1002/prot.21690
66.
BackusKM, CorreiaBE, LumKM, et al.Proteome-wide covalent ligand discovery in native biological systems. Nature, 2016; 534(7608):570–574; doi: 10.1038/nature18002
67.
GreerEL, ShiY. Histone methylation: A dynamic mark in health, disease and inheritance. Nat Rev Genet, 2012; 13(5):343–357; doi: 10.1038/nrg3173
68.
MattiroliF, SixmaTK. Lysine-targeting specificity in ubiquitin and ubiquitin-like modification pathways. Nat Struct Mol Biol, 2014; 21(4):308–316; doi: 10.1038/nsmb.2792
69.
ChoudharyC, WeinertBT, NishidaY, VerdinE, MannM. The growing landscape of lysine acetylation links metabolism and cell signalling. Nat Rev Mol Cell Biol, 2014; 15(8):536–550; doi: 10.1038/nrm3841
70.
WeerapanaE, WangC, SimonGM, et al.Quantitative reactivity profiling predicts functional cysteines in proteomes. Nature, 2010; 468(7325):790–795; doi: 10.1038/nature09472
71.
HackerSM, BackusKM, LazearMR, et al.Global profiling of lysine reactivity and ligandability in the human proteome. Nat Chem, 2017; 9(12):1181–1190; doi: 10.1038/nchem.2826
72.
AbbasovME, KavanaghME, IchuTA, et al.A proteome-wide atlas of lysine-reactive chemistry. Nat Chem, 2021; 13(11):1081–1092; doi: 10.1038/s41557-021-00765-4
