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1.
Protein Expr Purif ; 180: 105818, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33418060

RESUMEN

Campylobacteriosis is a disease in humans caused by the infection from Campylobacter spp. Human cases are mainly due to Campylobacter jejuni, although C. coli can cause gastroenteritis in humans as well. The bacteria are commensal in chicken tract and can be contaminated into chicken products during processing. Obviously, detecting reagents such as a specific antibody is essential for the development of immune-based detection methods for C. jejuni or C. coli. In this study, in silico techniques were used to design a chimeric recombinant antigen, named multiepitope antigen (MEA), for the production of specific polyclonal antibody. To design MEA polypeptide based on C. jejuni fibronectin-binding protein or CadF, four conserved and unique antigenic peptides were identified and fused together directly. The C. jejuni CadF-based MEA polypeptide fused with two single six-histidine tags at both C- and N-terminal ends was expressed under Escherichia coli expression system. The recombinant MEA was successfully produced and purified by Ni-NTA resin with a high satisfactory yield. Indirect ELISA results showed that anti-MEA polyclonal antibody derived from rabbit serum had a titer of 16,000, indicating high antigenicity of MEA polypeptide. Dot blot results also confirmed that the produced anti-MEA antibody could specifically recognize both C. jejuni and C. coli whole cells as expected while there was no cross-reactivity to non-Campylobacter spp. tested in this study.


Asunto(s)
Anticuerpos Antibacterianos/inmunología , Antígenos Bacterianos , Proteínas de la Membrana Bacteriana Externa , Campylobacter coli , Campylobacter jejuni , Proteínas Portadoras , Epítopos , Expresión Génica , Proteínas Recombinantes de Fusión , Animales , Anticuerpos Antibacterianos/química , Antígenos Bacterianos/biosíntesis , Antígenos Bacterianos/química , Antígenos Bacterianos/genética , Antígenos Bacterianos/inmunología , Proteínas de la Membrana Bacteriana Externa/biosíntesis , Proteínas de la Membrana Bacteriana Externa/química , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/inmunología , Campylobacter coli/química , Campylobacter coli/genética , Campylobacter coli/inmunología , Campylobacter jejuni/química , Campylobacter jejuni/genética , Campylobacter jejuni/inmunología , Proteínas Portadoras/biosíntesis , Proteínas Portadoras/química , Proteínas Portadoras/genética , Proteínas Portadoras/inmunología , Epítopos/biosíntesis , Epítopos/química , Epítopos/genética , Epítopos/inmunología , Conejos , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología
2.
J Microbiol Biotechnol ; 33(6): 771-779, 2023 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-36959178

RESUMEN

Biofilms are a significant concern in the food industry. The utilization of plant-derived compounds to inactivate biofilms on food contact surfaces has not been widely reported. Also, the increasing negative perception of consumers against synthetic sanitizers has encouraged the hunt for natural compounds as alternatives. Therefore, in this study we evaluated the antimicrobial activities of ethanol extracts, acetone extracts, and essential oils (EOs) of seven culinary herbs against Salmonella enterica serotype Typhimurium and Listeria innocua using the broth microdilution assay. Among all tested extracts and EOs, the ethanol extract of Piper betle L. exhibited the most efficient antimicrobial activities. To evaluate the biofilm inactivation effect, S. Typhimurium and L. innocua biofilms on pitted and smooth stainless steel (SS) coupons were exposed to P. betle ethanol extract (12.5 mg/ml), sodium hypochlorite (NaClO; 200 ppm), hydrogen peroxide (HP; 1100 ppm), and benzalkonium chloride (BKC; 400 ppm) for 15 min. Results showed that, for the untreated controls, higher sessile cell counts were observed on pitted SS versus smooth SS coupons. Overall, biofilm inactivation efficacies of the tested sanitizers followed the trend of P. betle extract ≥ BKC > NaClO > HP. The surface condition of SS did not affect the biofilm inactivation effect of each tested sanitizer. The contact angle results revealed P. betle ethanol extract could increase the surface wettability of SS coupons. This research suggests P. betle extract might be utilized as an alternative sanitizer in food processing facilities.


Asunto(s)
Antiinfecciosos , Listeria monocytogenes , Piper betle , Acero Inoxidable/análisis , Acero Inoxidable/farmacología , Microbiología de Alimentos , Biopelículas , Etanol/farmacología , Salmonella typhimurium , Antiinfecciosos/farmacología , Recuento de Colonia Microbiana
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