Ligilactobacillus salivarius KC27L'den Elde Edilen Hücresiz Süpernatant (CFSKC27L) ve Ekzopolisakkaritin (EPSKC27L) Antioksidan ve Antimikrobiyal Aktivitelerinin Belirlenmesi

Year 2024, , 928 - 941, 20.09.2024

Kübra Çelik , Zehranur Yuksekdag , Berat Çınar Acar , Filiz Kara

https://doi.org/10.33462/jotaf.1370839

Abstract

Ankara bölgesinden (Türkiye) alınan kanatlı dışkılarından biyokimyasal yöntemlerle elde edilen sonuçlara göre Lactobacillus cinsine ait 26 laktik asit bakterisi izole edilmiştir. Bu çalışma, bu izolatları ekzopolisakkarit (EPS) üretimi açısından taramıştır. Bu izolatlarda 8 mg L-1 ila 353 mg L-1 arasında değişen EPS üretimi tespit edilmiştir. Bundan sonra yapılacak çalışmalar için en yüksek EPS üreten izolat (KC27L) seçilmiştir. İzolat 16S rRNA analizi ile Ligilactobacillus salivarius olarak tanımlanmıştır. Ayrıca, yüksek EPS üretimi sergileyen KC27L suşuna (EPSKC27L) ait hücresiz süpernatantın (CFSKC27L) ve farklı konsantrasyonlardaki (0.5 mg L-1 ve 1 mg L-1) EPS'nin anti-biyofilm ve antioksidan yetenekleri belirlenmiştir. CFSKC27 ve farklı konsantrasyonlarda (0.5 ve 1 mg mL-1) EPSKC27L, Escherichia coli ATCC 11229, Enterococcus faecalis ATCC 29212 ve Staphylococcus aureus EB-1 üzerinde anti-biyofilm etkisini belirlemiştir. 1 mg mL-1 EPSKC27L'de en yüksek anti-biyofilm etkisi % 87 inhibisyon ile E. coli ATCC 11229'da tespit edilmiştir. Üç farklı yöntem ile (1.1-Difenil-2-pikrilhidrazil radikali (DPPH) giderim etkisi, Fe2+ iyon şelatlama aktivitesi ve süperoksit anyon radikal temizleme aktivitesi) antioksidan aktivite belirlenmiştir. En yüksek 1.1-Difenil-2-pikrilhidrazil radikali (DPPH) giderim etkisi, Fe2+ iyon şelatlama aktivitesi ve süperoksit anyon radikal temizleme aktivitesi 1 mg mL-1 EPSKC27L'de (sırasıyla %79.6, %24.9 ve %61.6) bulunmuştur. 1 mg mL-1 EPSKC27L'nin hem anti-biyofilm hem de antioksidan aktiviteleri postbiyotikten daha yüksek çıkmıştır. Son olarak EPSKC27L'nin kısmi saflaştırılmasının ardından moleküler karakterizasyonu yapılmıştır. EPSKC27L, molekür ağırlıkları 1.6x103 ve 6.4 x104 Da olan iki fraksiyona sahiptir. EPSKC27L’nin monosakarit bileşenleri glikoz (%53.1), fruktoz (%18.5), arabinoz (%14.6) ve mannoz (%13.8) olarak bulunmuştur. L. salivarius'tan elde edilen CFSKC27L ve EPSKC27L antioksidan ve anti-biyofilm ajanları olabilir.

Keywords

Ligilactobacillus salivarius, Hücre içermeyen supernatant, Ekzopolisakkarit, Anti-biyofilm, Antioksidan

Supporting Institution

This work is supported by the Gazi University Scientific Research Projects Department Research Project (Project No: 05/2017-08)

Project Number

05/2017-08

Determination of Antioxidant and Antimicrobial Activities of Cell-Free Supernatant (CFSKC27L) and Exopolysaccharide (EPSKC27L) obtained from Ligilactobacillus salivarius KC27L

Abstract

Twenty-six lactic acid bacteria were obtained from poultry feces sampled located in the Ankara area (Türkiye) and belong to the Lactobacillus genus according to the results obtained by biochemical methods. This study screened these isolates for exopolysaccharides (EPS) production. EPS production was detected in these isolates, varying from 8 mg L-1 to 353 mg L-1. The highest EPS-producing isolate (KC27L) was selected for further studies. The isolate was identified as Ligilactobacillus salivarius by 16S rRNA analysis. Furthermore, the anti-biofilm and antioxidant abilities of the cell-free supernatant (CFSKC27L) and different concentrations (0.5 mg L-1 and 1 mg L-1) of EPS belonging to the KC27L strain (EPSKC27L) that exhibited high EPS production were determined. CFSKC27 and different concentrations (0.5 mg L-1 and 1 mg mL-1) of EPSKC27L determined the anti-biofilm impact on Escherichia coli ATCC 11229, Enterococcus faecalis ATCC 29212, and Staphylococcus aureus EB-1. The highest anti-biofilm effect in 1 mg mL-1 EPSKC27L was detected at E. coli ATCC 11229 with 87 % inhibition. Three different methods (1.1-Diphenyl-2-picrylhydrazyl radical (DPPH) removal impact, Fe2+ ion chelating and superoxide anion radical scavenging activity) designated antioxidant activity. The highest 1.1-Diphenyl-2-picrylhydrazyl radical (DPPH) removal impact, Fe2+ ion chelating, and superoxide anion radical scavenging activity were found in 1 mg mL-1 EPSKC27L (79.6%, 24.9%, and 61.6%, respectively). Both anti-biofilm and antioxidant activities of 1 mg mL-1 EPSKC27L were higher than postbiotic. Finally, its molecular characterization was done following the partial purification of the EPSKC27L. The EPSKC27L has two fractions with molecular weights of 1.6x103 and 6.4 x104 Da. Monosaccharide components of EPSKC27L were found to be glucose (53.1%), fructose (18.5%), arabinose (14.6%) and mannose (13.8%). CFSKC27L and EPSKC27L obtained from L. salivarius can be antioxidants and anti-biofilm agents.

Keywords

Ligilactobacillus salivarius, Cell-free supernatant, Exopolysaccharide, Anti-biofilm, Antioxidant

Supporting Institution

This work is supported by the Gazi University Scientific Research Projects Department Research Project (Project No: 05/2017-08)

Project Number

05/2017-08

Thanks

The authors would like to thank Gazi University Scientific Research Projects Department for funding the 05/2017-08 coded project including this study

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