African Journal of Biotechnology Vol. 10(22), pp. 4518-4523, 30 May, 2011 Available online at http://www.academicjournals.org/AJB DOI: 10.5897/AJB10.2201 ISSN 1684–5315 © 2011 Academic Journals Review Ellagic acid: Biological properties and biotechnological development for production processes Leonardo Sepúlveda1, Alberto Ascacio1, Raúl Rodríguez-Herrera1, Antonio Aguilera-Carbó2 and Cristóbal N. Aguilar1* 1Department of Food Science and Technology, Universidad Autónoma de Coahuila, Saltillo, Coahuila, México. 2Department of Food Science and Nutrition, Universidad Autónoma Agraria Antonio Narro, Buenavista, Saltillo, Coahuila, México. Accepted 15 April, 2011 Ellagic acid, 2,3,7,8-tetrahydroxy-chromeno[5,4,3-cde]chromene-5,10-dione, is a powerful bioactive compound with many potential pharmacological and industrial applications. In this review, the chemical aspects, biological properties and diverse potential applications of ellagic acid for different industries were described. This review also discussed the advance in ellagitannin biodegradation, focusing on the process of isolation of microorganisms and strain selection, medium and culture optimization, as well as fermentation systems for commercially viable industrial scale production. The performances of various fermentation techniques that have been applied for the production of ellagic acid from residual by-products were compared, while the advantages and disadvantages of each plant source were also discussed. Key words: Ellagic acid, ellagitannin, biodegradation, fungal physiology, solid-state fermentation, submerged fermentation. INTRODUCTION Tannins are polyphenolic compounds present in different Also, these bio-active molecules offer protection against plant species, they are water soluble with molecular masses ruminants due to the formation of complexes between ranging form 500 to 20000 Da (Khanbabaee and Ree, plant tannins and animal proteins such as hydroxyproline- 2001). They are distributed in fruits, stalks, peels, roots rich proteins. Formation of such complexes results in a and leaves (Haslam and Cai, 1994). Concentration and bitter and disagreeable sensation which deters potential kinds of these compounds are determined by plant predators. Molecules present in the plants that are specie. Tannins are considered as secondary metabolites susceptible to microbial degradation, such as proteins in plant physiology and are generally classified into four and polysaccharides, have evolved to become highly groups: gallotannins, ellagitannins, condensed tannins resistant to such degradation when linked to tannins and complex tannins (Khanbabae and Ree, 2001). (Aguilera-Carbo et al., 2009). These phytochemicals are located in the vacuoles of One of the most important roles of tannins is the intact plant cells, and are released upon attack by diverse growth inhibition towards many microorganisms including microorganisms, including viruses, bacteria and fungi, bacteria, yeasts and fungi. They are therefore recalcitrant thereby avoiding potential infection of plant tissues. In to enzyme degradation by most microorganisms. addition, the astringent properties of tannins are well Condensed tannins are more resistant to microbial attack known to stop the infestation of insects (Koide et al., than hydrolysable tannins and are more toxic for 1998). foodborne pathogens (Aguilera-Carbo et al., 2005). Tannins retard the decomposition of solid organic material through inhibition of degrading enzymes of attacking microorganisms. When tannins are complexed with microbial proteins or polysaccharides, the interactions *Corresponding author. E-mail: [email protected]. formed are often irreversible and this characteristics mx. Tel: 52 (844) 4161338. Fax: 52 (844) 415 9534. confer on them the bactericide and bacteriostatic proper- Sepúlveda et al. 4519 four phenolic groups and two lactones, which can act as hydrogen-forming sides and electron acceptors, respectively, and represent the hydrophilic domain (Aguilera-Carbó et al., 2005). Ellagic acid is a compound that has generated commercial interest in recent years due to its properties, applications and benefits to human health. BIOLOGICAL PROPERTIES OF ELLAGIC ACID Antioxidants are compound that can delay, inhibit or prevent the oxidation of compounds, trapping free radi- cals and reducing oxidative stress. The presence of ellagic acid in various commercial products giving anti- oxidant activity has also been reported. These molecules have a variety of benefits for their anti-mutagenic, antimicrobial and antioxidant properties, and inhibitors of human immunodeficiency virus (HIV) (Tatsuo et al., 1998; Feldman et al., 1999; Akiyama et al., 2001; Vattem and Shetty, 2003; Ruibal et al., 2003). Ellagic acid prevents Figure 1. Hexaydroxidiphenic group of the formation of various tumors, this mechanism of action ellagitannins. can be possible because compounds such as ellagi- tannins and ellagic acid, explicitly interact with the cells walls or sites with facility to complex proteins, preventing ties. However, some microorganisms tolerate the the proliferation of metastatic cells (Tatsuo et al., 1998). presence of tannins and/or use these com-pounds as Losso et al. (2004) evaluated the potential cytotoxic and carbon source. That ability is generated by the production anti-proliferative activities of ellagic acid in human cells, of a tannin-degrading enzyme or tannase, produced lung, colon, breast and prostate cancer, and showed that mainly by microorganisms of the genus Aspergillus and doses of 1 to 100 mol/L inhibited the proliferation of the Penicillium (Aguilar et al., 2008). cancers mentioned. It has been demonstrated that ellagic Ellagitannins (ETs) correspond to esters of ellagic acid acid acts, possibly causing apoptosis in cancer cells by with glucose or quinic acid. ETs are generated by inhibiting factors that promote metastasis. Huertz et al. oxidative interaction of gallotannins with at least 2 galoyl (2005) studied a coordinated response between a typical units, originating from a group called hexahydroxy- carcinogen such as benzo [] piren-7, 8-diol-epoxide dyphenic acid (HHDP) (Figure 1). This group is released (BPDE) and ellagic acid, which they see as chemopre- after hydrolysis of ETs, and spontaneously lactonized or ventive agent in some cancers caused by polycyclic rearranged to generate the ellagic acid. There are more aromatic hydrocarbons (PAHs). The authors proposed a than 500 ellagitannin structures reported (Feldman et al., mechanism by which ellagic acid, in the formation of 1999). The ellagitannins are mainly derived from the bark DNA, is adducted by the direct compaction of the of oak (Quercus spp.). Bianco et al. (1998) indicated ET's carcinogen. can be can be classified according to the number of Studies of ellagic acid in cancer cells have demon- HHDP groups in the molecule, as monomeric, oligomeric strated the induction of apoptosis or cell death, and polymeric structures (grade of polymerization). The preventing a continuous tumor growth (Ito et al., 1999). principal molecule for the formation or biosynthesis of This mechanism was proposed by Heather et al. (2007) ETs is pentagaloyl glucose (PGG). The oxidation of this when polyphenol-rich raspberry extracts were made and molecule is carried out enzymatically by plant polyphenol fractioned by chromatography on C18 solid phase, oxidase, which performs the coupling of galoyl groups to reporting that one of the separated compounds identified generate the group HHDP (Figure 2). as ellagic showed the ability to inhibit in vitro, the proliferation of human cervical cancer (HeLa) cells, and the possible mechanism explained was that ellagic acid ELLAGIC ACID from ETs induces apoptosis through intrinsic mito- chondrial pathway. Other studies have identified impor- Ellagic acid (Figure 3) is a molecule with a molecular tant ETs-type phytochemicals in diverse berry fruit weight of 302 gmol-1. It is a molecule that is highly extracts, which are characterized by biological properties thermostable (melting point of 350°C) (Ascacio et al., such as anticancer (colon, prostate and leukemia), anti- 2010), and has four rings representing the lipophilic domain, neuodegenarative, anti-viral, etc. (Mertens-Talcott et al., 4520 Afr. J. Biotechnol. Figure 2. Scheme of PGG oxidation (A) and the ellagitannin telimagrandin II (B) by a phenol oxidase type enzyme (Niemetz et al., 2003). aureus strains (Machado et al., 2002). These biological properties have been attributed to the ETs ability to inhibit gyrase activity which is associated with the cleavage of the DNA strand during replication process (Weidner- Wells et al., 1998). It is pointed out that this bacterium does not have the ability to resist this type of antibacterial action. Also, different concentrations of ellagic acid against the human immunodeficiency virus have been tested, and it demonstrated the ability to inhibit the virus growth and spread due to the ability of ellagic acid to attach to HIV proteins (Notka et al., 2004). Also, viruses do not have the ability to replicate themselves in the presence of ETs; because they inhibit the integrase enzyme involved in the insertion of viral genomic material to host cells (Jegede et al., 2008). This is particular in the Figure 3. Structure of ellagic acid. case of retroviruses, including HIV, were these very specific phytochemicals also inhibit reverse transcriptase activity (Aguilera-Carbó et al., 2009). Therefore, there is 2003; Seeram et al., 2006). Atta-Ur-Rahman