Send Orders for Reprints to [email protected] Mini-Reviews in Organic Chemistry, 2014, 11, 15-27 15 Purification and Characterization of Venom Components as Source for Antibiotics Leidy Johana Vargas Muñoza* and Sebastián Estrada-Gómeza,b aPrograma de Ofidismo/Escorpionismo, Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Medellín, Colombia., Calle 70 # 52-2, A.A. 1226, Medellín, Colombia; bFacultad de Química Farmacéutica, Universidad de Antioquia UdeA, Medellín, Colombia Abstract: The extensive use of antibiotics in medicine, food industry, and agriculture has led to a frequent emergence of multidrug-resistant bacteria, which creates an urgent need for new antibiotics. It is now widely recognized that venom proteins could play a promising role against multidrug-resistant bacteria. Different proteins with antibacterial activity have been characterized from the venoms of snakes, spiders and scorpions in the last decade. This review summarizes the proteins and peptides that have been purified and characterized from different venoms with antibacterial activity. Keywords: Antimicrobial activity, snake, spider, scorpion, venom, peptides. 1. INTRODUCTION During the last decade, diverse proteins with antibacterial activity have been characterized from different sources like Bacterial infections are among the 10 leading causes of plants, animals, mammals, microorganisms and venoms death worldwide according to the World Health Organization (snake, scorpion and spider venoms) being the last but the [1]. The presence and current emergence of multiple- main focus in this review. resistant strains make the risk of these infections more threatening since a treatment becomes unreachable. In fact, bacterial resistance has been the principal aspect responsible 2. SNAKE VENOMS for increasing morbidity, mortality and health care costs of Snake venoms are cocktails of enzymatic and non- bacterial infections [2]. Therefore, research for new antimi- enzymatic proteins used for both immobilization and pre- crobials or antibacterial prototypes is continuously necessary digestion of prey. The most common enzymes present in for drug design and development seeking the treatment of these venoms include acetylcholinesterases, L-amino acid infections involving multidrug-resistant microorganisms [1, oxidases, desintegrins, serine proteinases, metalloprotein- 3] It is also of considerable interest to explore and develop ases, lectins and phospholipases A2 [6]. These cocktails have antimicrobials with a new mechanism(s) of action which can been tested against different microorganism; from the Vi- potentially avoid the appearance of drug resistance. Nature is peridae, the experimental data revealed that the venoms from a wide source of antimicrobial peptides since they are ubiq- Agkistrodon rhodostoma, Bothrops atrox and B. jararaca uitous as part of the innate immune system and host defense exhibited a promising antibacterial activity against some of mechanisms. They have been increasingly recognized as a the Gram-positive and Gram-negative bacteria like Entero- critical first line of defense against many pathogens isolated coccus faecalis, Staphylococcus epidermidis and Staphylo- from various sources. coccus aureus [7]; while venoms from Crotalus viridus Antimicrobial resistance is a natural phenomenon which helleri, C. atrox and C. horridus horridus inhibited the entails an inherent risk, associated to antimicrobial drugs growth only from aerobic species like S. epidermidis, Pseu- use, comprising the intrinsic and acquired resistance. The domona aeruginosa, and Enterobacter cloacae [8]. Ciscotto intrinsic is referred to a non-genetic related resistance [4], et al. [9], tested B. jararacussu crude venom against differ- while the acquired resistance, is achieved by antibiotics use, ent bacteria. It was active against Gram-positive bacteria and occurs by mutations in the bacterial genome or by hori- such as Eubacterium lentum, Peptostreptococcus anaero- zontal transference of genetic information [4]. The most im- bius, Propionibacterium acnes, S. aureus and S. epidermidis, portant antibiotics resistance mechanisms are related with the and Gram-negative bacteria such as Porphyromonas gin- antibiotic drawing diminution, increase of antibiotic efflux, givalis, Prevotella intermedia, P. aeruginosa, and Salmo- inactivation or modification of antibiotic targets, hydrolysis nella typhimurium. Antimicrobial activity of this venom, was or any chemical modification of the antibiotic structure and associated with the presence of proteins like LAAO and/or any new resistance acquired by horizontal genetic transmis- phospholipase A2 due to variation in content and quality of sion [4, 5]. these proteins in snake venom. Moreover, the Australian elapid Pseudechis australis inhibited S. aureus and Es- cherichia coli [10]. *Address correspondence to this author at the Calle 70 # 52-2, Medellín Colombia, zip code: 050010. Medellin, Colombia; Tel: (0574) 219 6649; Several toxins from these venoms have been isolated and Fax: (0574) 2631914; E-mail: [email protected] characterized, especially L- amino acid oxidases (LAAOs) 1875-6298/14 $58.00+.00 © 2014 Bentham Science Publishers 16 Mini-Reviews in Organic Chemistry, 2014, Vol. 11, No. 1 Muñoz and Estrada-Gómez and phospholipase A2 (PLA2), enhancing antimicrobial activ- and physiochemical properties, inactivation by pH changes ity as seen below in this review. or freezing and others [12]. Now, LAAOs, have emerged as an interesting object of study for their enzymology, struc- 2.1. LAAOs tural biology, pharmacology and because of their relatively L-amino acid oxidases (LAAOs, E.C.1.4.3.2) are enanti- easy purification. Recently, more and more SV-LAAOs have been characterized with distinct molecular mass, substrate oselective flavoenzymes catalyzing the stereospecific oxida- preference, interactions with platelets [11c, 13], induction of tive deamination of a wide range of L-amino acids to form - hemorrhage [11d] and apoptosis [14], equally antileishma- keto acids, ammonia and hydrogen peroxide (H2O2). LAAOs have been reported in Viperidae, Crotalidae and Elapidae nial [15] and antibacterial activities [10]. snakes, usually as FAD-(flavin adenine dinucleotide) or First reports about LAAO antimicrobial activity came in FMN- (flavin mononucleotide) homodimeric binding glyco- the early 70s when Skarnes et al. [16] reported the antimi- proteins. Flavins in LAAOs are responsible for the yellow crobial activities elicited by C. adamanteus LAAOs. However, color of snake venom and contribute to toxicity via the oxi- since then few LAAOs were isolated till 1990 and later in dative stress arising from the production of H2O2 [9, 11]. year 2000 when the interest increased in these proteins lead- Before 1990s, the studies in the field of snake venom ing to a rise in their isolation and characterization. (Table 1) LAAOs (SV-LAAOs) used to be focused on their enzymatic summarizes SV-LAAOs reported with antimicrobial activity. Table 1. LAAOs Isolated with Antibacterial Activity. Gram (G) Negative (-) and Positive (+) LAAO Isolated from Snake Venom Antimicrobial Activity Properties References Crotalus adamanteus Active against G() P. aeruginosa y E. coli Optimum pH 6.5-7.0 Skarnes [16] More active against G(+) Staphylococcus mutans, than C. durissus cascavella 120 kDa, dimer, pI 5.4 Toyama et al. [22] G() Xanthomonas axonopodis pv passiflorae Agkistrodon halys Active against G() E. coli and G(+) Bacillus subtilis 60.7 kDa, optimum pH: 8.8 Zhang et al. [53] Bothrops alternatus Active against G() E. coli, and G(+) B. subtilis 123 kDa, homodimer, pI 5.37 Stabeli et al. [11d] Active against G(+) S. aureus, G() P. aeruginosa and B. marajoensis 67 kDa, monomer, acidic Torres et al. [54] Candida albicans. Active against G() E. coli, Salmonella typhimurium, P. B. moojeni 140 kDa, homodimer; pI 4.8 Stabeli et al. [21] aeruginosa and G(+) S. aureus B. pauloensis More active against G() E. coli than G(+) S. aureus. 65 kDa, homodimeric, pI 6.3 Rodrigues et al. [55] B. pirajai Active against G() E. coli and P. aeruginosa 130 kDa, homodimer; pI 4.9 Izidoro et al. [56] Active against K. pneumoniae, P. aeruginosa and P. B. mattogrosensis 60 kDa, homodimer. Okubo et al., [25] mirabilis, Protobothrops Active against G(+) B. megaterilum and S. aureus, G() 110 kDa, homodimer Lu et al. [11c] (Trimeresurus) jerdonii E. coli and P. aeruginosa More active against G(+) S. aureus than G() E. coli Trimeresurus mucrosquamatus 110 kDa, homodimer Wei et al. [11e] and B. dysenteriae. Vipera lebetina More active against G() E. coli than G(+) B. subtilis. 140 kDa, homodimer, pI 5.3 Tonismagi et al. [23] Daboia russellii Active against G(+) S. aureus; G() P. aeruginosa and 100 kDa, homodimer; pI 8.8 Zhong et al. [24] siamensis E. coli. Naja naja oxiana More active against G(+) B. subtilis than G() E. coli 110 kDa, homodimer Zhong et al. [24] (Naja oxiana) Active against Aeromonas sp and G(+) B. subtilis and S. Pseudechis australis 142 kDa, dimer, 56 kDa Stiles et al. [10] aureus. A. hydrophila Active against gram-positive and gram-negative three isoforms: 80, 60.8 and Bothrops jararaca Ciscotto et al. [9] bacteria. 48.1 kDa, monomer 124.4 kDa, homodimer, pI Agkistrodon blomhoffii ussurensis Active against S. aureus. Sun et al. [57] 4.7 Active against P. aeruginosa, K. pneumoniae and Ophiophagus hannah 135 kDa, homodimer, pI 4.5 Lee et al. [58] E. coli. Agkistrodon halys pallas Active against E. coli. 60.7 kDa, pI 8.8 Liu et al., [59] Purification