Biocatalytic synthesis, antimicrobial properties and toxicity studies of arginine derivative surfactants
- 359 Downloads
Two novel arginine-based cationic surfactants were synthesized using as biocatalyst papain, an endopeptidase from Carica papaya latex, adsorbed onto polyamide. The classical substrate N α-benzoyl-arginine ethyl ester hydrochloride for the determination of cysteine and serine proteases activity was used as the arginine donor, whereas decyl- and dodecylamine were used as nucleophiles for the condensation reaction. Yields higher than 90 and 80 % were achieved for the synthesis of N α-benzoyl-arginine decyl amide (Bz-Arg-NHC10) and N α-benzoyl-arginine dodecyl amide (Bz-Arg-NHC12), respectively. The purification process was developed in order to make it more sustainable, by using water and ethanol as the main separation solvents in a single cationic exchange chromatographic separation step. Bz-Arg-NHC10 and Bz-Arg-NHC12 proved antimicrobial activity against both Gram-positive and Gram-negative bacteria, revealing their potential use as effective disinfectants as they reduced 99 % the initial bacterial population after only 1 h of contact. The cytotoxic effect towards different cell types of both arginine derivatives was also measured. Bz-Arg-NHCn demonstrated lower haemolytic activity and were less eye-irritating than the commercial cationic surfactant cetrimide. A similar trend could also be observed when cytotoxicity was tested on hepatocytes and fibroblast cell lines: both arginine derivatives were less toxic than cetrimide. All these properties would make the two novel arginine compounds a promising alternative to commercial cationic surfactants, especially for their use as additives in topical formulations.
KeywordsArginine-based surfactants Papain Biocatalysis Antimicrobial activity Cytotoxic effect
This research was supported by projects PIP 0150 (CONICET), X-613 (UNLP), CTQ2012-31605 and BIO2013-44973-R (MINECO, Spain). Mass spectra were performed in the Unidad de Microanálisis y Métodos Físicos Aplicados a la Química Orgánica (UMYMFOR), CONICET-FCEN-UBA, Buenos Aires, Argentina. The valuable contribution of Dr. Alicia S. Cánepa (LADECOR, Depto. de Química, Fac. Cs. Exactas, UNLP) in the analysis of NMR spectra is also acknowledged. MEF was awarded CONICET fellowship. GLG and SRM are members of CONICET Researcher Career.
Conflict of interest
The authors declare that they have no conflict of interest.
- Arnon R (1970) Papain. In: Perlmann GE, Lorand L (eds) Methods in Enzymology, vol 19., Proteolytic EnzimesAcademic Press, New York, pp 226–244Google Scholar
- Benavides T, Mitjans M, Martínez V et al (2004) Assessment of primary eye and skin irritants by in vitro cytotoxicity and phototoxicity models: an in vitro approach of new arginine-based surfactant-induced irritation. Toxicology 197:229–237. doi: 10.1016/j.tox.2004.01.011 PubMedCrossRefGoogle Scholar
- Castillo-Expósito JA (2006) Studies on antimicrobial activity of arginine-based surfactants and chemo- enzymatic synthesis of novel amphiphiles based on l-arginine and d-fagomine. Ph D Thesis, Universidad Autónoma de BarcelonaGoogle Scholar
- Chimutengwende-Gordon M, Pendegrass C, Bayston R, Blunn G (2014) Preventing infection of osseointegrated transcutaneous implants: incorporation of silver into preconditioned fibronectin-functionalized hydroxyapatite coatings suppresses Staphylococcus aureus colonization while promoting viable fibroblast growth in vitro. Biointerphases 9:031010. doi: 10.1116/1.4889977 PubMedCrossRefGoogle Scholar
- Clapés P, Morán C, Infante MR (1999) Enzymatic synthesis of arginine-based cationic surfactants. Biotechnol Bioeng 63:333–343. doi: 10.1002/(SICI)1097-0290(19990505)63:3<333:AID-BIT10>3.0.CO;2-G PubMedCrossRefGoogle Scholar
- Florence AT, Attwood D (2006) Physicochemical principles of pharmacy, 4th edn. Pharmaceutical Press, LondonGoogle Scholar
- Messing RA (1975) Introduction and general history of immobilized enzymes. In: Messing RA (ed) Immobilized enzymes for industrial reactors. Academic Press, New York, pp 2–3Google Scholar
- Mitin Y, Braun K, Kuhl P (1997) Papain catalyzed synthesis of glyceryl esters of N-protected amino acids and peptides for the use in trypsin catalyzed peptide synthesis. Biotechnol Bioeng 54:287–290. doi: 10.1002/(SICI)1097-0290(19970505)54:3<287:AID-BIT9>3.0.CO;2-B PubMedCrossRefGoogle Scholar
- Nogueira DL, Mitjans M, Infante MR, Vinardell MP (2011) Comparative sensitivity of tumor and non-tumor cell lines as a reliable approach for in vitro citotoxicity screening of lysine-based surfactants with potential pharmaceutical applications. Int J Pharm 420:51–58. doi: 10.1016/j.ijpharm.2011.08.020 PubMedCrossRefGoogle Scholar
- Piera E, Infante MR, Clapés P (2000) Chemo-enzymatic synthesis of arginine-based gemini surfactants. Biotechnol Bioeng 70:323–331. doi: 10.1002/1097-0290(20001105)70:3<323:AID-BIT9>3.0.CO;2-N PubMedCrossRefGoogle Scholar