Peptides DLL37-1 and LL37-1, an alternative to inhibit biofilm formation in clinical isolates of <i>Staphylococcus aureus </i> and Staphylococcus epidermidis

ALBA, MARYI LORENA SEGURA; DURÁN-RODRIGUEZ, ANDREA TATIANA; PULIDO, LUZ MARY SALAZAR; ESCOBAR-PÉREZ, JAVIER; GUTIÉRREZ, SERGIO ALEJANDRO; OSPINA, JEANNETTE NAVARRETE; BERMÚDEZ, GLADYS PINILLA; MOLINA, LILIANA CONSTANZA MUÑOZ

doi:10.1590/0001-3765202220210848

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Sumário

HEALTH SCIENCES • An. Acad. Bras. Ciênc. 94 (3) • 2022 • https://doi.org/10.1590/0001-3765202220210848 linkcopy

Peptides DLL37-1 and LL37-1, an alternative to inhibit biofilm formation in clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis

Authorship SCIMAGO INSTITUTIONS RANKINGS

Staphylococcus aureus and Staphylococcus epidermidis have microbial surface components recognizing adhesive matrix molecules (MSCRAMM) adhesion proteins that enhance their biofilm formation ability, as well as virulence factors that influence morbidity and mortality in hospital settings. In this work, four peptides analogous of the peptide LL-37 that were evaluated to inhibit biofilm formation and its action potential on the expression of MSCRAMM proteins in clinical isolates through different tests, such as crystal violet, PCR and qPCR. In total, 96.8% of S. aureus were strong in biofilm formation in contrast to 48.4% of S. epidermidis. sdrG and sdrF genes were present in 100% of S. epidermidis strains and in all isolates. In S. aureus, specific genes that code for MSCRAMM proteins were detected: clfA (89%), clFB, sdrC and fnBA (94%). The peptides did not show hemolytic or cytotoxic activity. In this study, it was evidenced that of the peptides DLL37-1 at a 5 µM concentration was an efficacious antimicrobial agent and depicted greater biofilm inhibition in both bacterial species. Exhibiting a significant inhibition rate in S. aureus, this peptide caused a negative regulation in the expression of the genes clfA and sdrC, showed greater biological activity.

Key words
Biofilms; MSCRAMM; Peptides; S. epidermidis

Name	Sequence
LL37 original	L G D F R K S K E K I G K E F K R I V Q R I K D F L R N L V P R T E S
LL37-1	G R K S A K I G K R A K R I V Q R I K D F L R
LL37-1	Free Amino (C-terminal) Amidation
ACLL37-1	F -R K S K E K I G K E F K R I V Q R I K D F L R
ACLL37-1	(N-terminal) Acetylation (C-terminal) Amidation
ACLL37-2	G R K S A K I G K R A K R I V Q R I K D F L R
ACLL37-2	(N-terminal) Acetylation (C-terminal) Amidation
DLL37-1	(d-PHE) G R K S A K I G K R A K R I V Q R I K D (d-F)L R
DLL37-1	(N-terminal) Acetylation (C-terminal) Amidation

Name	Sequence	Length	Tm	%GC	Size pb	Gene
clfA-F	TGCTGCACCTAAAACAGACG	20	60.05	50.00	155	clfA
clfA-R	CAGTTACCGGCGTTTCTTCC	20	61.89	55.00	155	clfA
clfB-F	AGCATTTACTACCGGTTCAGCA	22	60.03	45.45	96	clfB
clfB-R	CCACAAACGATTTCCAATGCGC	22	61.79	50.00	96	clfB
sdrC-F	CCGTTTCTACTGGCAATGCT	20	60.27	50.00	148	sdrC
sdrC-R	GTTACATTGCCAACGCCTTT	20	60.00	45.00	148	sdrC
fnbA-F	TACCGCTATTTTGGCCACCT	20	61.71	50.00	180	fnbA
fnbA-R	AGCATGGTCAAGCACAAGGA	20	61.71	50.00	180	fnbA
sdrG-F	TCGCCATCAACTAACGCTCAA	21	60.34	47.62	188 pb	sdrG
sdrG-R	CCCTGTTCAGCAAGTTCGTC	20	59.13	55.00	188 pb	sdrG
sdrF-F	GGACGATTACACGACGATACAGC	23	62.97	52.17	137 pb	sdrF
sdrF- R	CAAGTCACCTTGTCCTTGACCT	23	64.01	52.17	137 pb	sdrF

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[tfnI] Underlined: conserved native LL37 sequences. Modified amino acids.