FBP-435; No.

of Pages 7

ARTICLE IN PRESS
food and bioproducts processing x x x ( 2 0 1 3 ) xxxxxx

Contents lists available at ScienceDirect

Food and Bioproducts Processing

journal homepage: www.elsevier.com/locate/fbp

Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation
Juan Buenrostro-Figueroa a , Alberto Ascacio-Valds a , Leonardo Seplveda a , Reynaldo De la Cruz a , Arely Prado-Barragn b , Miguel A. Aguilar-Gonzlez c , Ral Rodrguez a , Cristbal N. Aguilar a,
a b c

Food Research Department, School of Chemistry, Universidad Autnoma de Coahuila, Saltillo, 25280, Coahuila, Mexico Department of Biotechnology, Universidad Autnoma Metropolitana, Iztapalapa, 09340, Mexico CINVESTAV-IPN, Ramos Arizpe, 25900, Coahuila, Mexico

a b s t r a c t
Ellagitannase is a novel enzyme responsible for biodegradation of ellagitannins and ellagic acid production. Ellagic acid is a bioactive compound with great potential in food, pharmaceutical and cosmetic industries. This work describes the ellagitannase enzyme production from partial puried ellagitannins as inducers by Aspergillus niger GH1 grown on solid-state fermentation. Solid-state fermentation was carried out on four different lignocellulosic materials (sugarcane bagasse, corn cobs, coconut husks and candelilla stalks) as matrix support and production of ellagitannase enzyme was evaluated. All lignocellulosic materials were characterized in terms of water absorption index and critical humidity point. The best lignocellulosic materials for ellagitannase production were sugarcane bagasse and corn cobs (1400 U L1 and 1200 U L1 , respectively). The lowest values were obtained with candelilla stalks (500 UL-1). The highest specic productivity was obtained with corn cobs (2.5 U mg1 h1 ) which enable increase ellagitannase productivity up to 140 times. Corn cobs have great potential as support matrix for production of fungal ellagitannase in SSF. 2013 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved. Keywords: Ellagitannins; Ellagic acid; Biodegradation; Agro industrial by-products; Aspergillus niger GH1; Corn cobs

1.

Introduction

Elagitannins are an important group of phytochemical compounds with great value in food, pharmaceutical and cosmetic industries. The ellagitannins (ETs) are watersoluble hydrolysable polyphenolic compounds (Wilson and Hagerman, 1990). These are considered secondary plant metabolites found in cytoplasm and cell vacuoles (Khadem and Marles, 2010). ETs presence has been principally described in leaves, stalks, husks of some fruits, owers, etc. (AscacioValds et al., 2011). When ETs are exposed to acidic or basic strong conditions, the ester bounds are hydrolyzed and the hexahydroxydyphenic acid group (HHDP) is released, which spontaneously rearranged to form a stable and insoluble

dilactone, commonly named ellagic acid (Aguilera-Carb et al., 2008b; Gross, 2009) Recent studies on sources and biological properties of ellagic acid showed its relevant bioactivity, such as antioxidant, anti-inammatory, antiviral, antimicrobial, antimutagenic, antitumoral and anticarcinogenic, among others (Ascacio-Valds et al., 2011). Ellagic acid is present in considerable amounts in cranberry (Vattem and Shetty, 2003), raspberry (Koponen et al., 2007) and pomegranate fruits (Aguilera-Carb et al., 2008a; Robledo et al., 2008; Seeram et al., 2005). Several authors have reported the recovery of high levels of ETs with a high purity degree from pomegranate husks (Ascacio-Valds et al., 2010; Robledo et al., 2008; Seeram et al., 2005).

Corresponding author. Tel.: +52 844 416 1238; fax: +53 844 415 9534. E-mail address: cristobal.aguilar@uadec.edu.mx (C.N. Aguilar). Received 3 January 2013; Received in revised form 8 May 2013; Accepted 18 August 2013 0960-3085/$ see front matter 2013 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.fbp.2013.08.010
Please cite this article in press as: Buenrostro-Figueroa, J., et al., Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation. Food Bioprod Process (2013), http://dx.doi.org/10.1016/j.fbp.2013.08.010

FBP-435; No. of Pages 7

ARTICLE IN PRESS
food and bioproducts processing x x x ( 2 0 1 3 ) xxxxxx

The studies on ETs hydrolysis have permitted established some control parameters of ellagic acid production using strong acids or bases, however this method generate high costs and high volumes of chemical wastes. Information about ETs biodegradation is scarce and confuse. However, in recent years some enzymatic studies have offered some clues to elucidate the degradation pathway of ETs. Action of some enzymes, such as -glucosidase (Vattem and Shetty, 2002, 2003), tannase, polyphenol oxidase (Shi et al., 2005), cellulase (Huang et al., 2007), valonea tannin hydrolase (Huang et al., 2005) and ellagitannase (Aguilera-Carb et al., 2009) has been reported. Microbial hydrolysis of ETs using enzymes has been poorly evaluated in SSF, and the rst reports hypothesized a synergistic action of different enzymes (Aguilera-Carb et al., 2008b). Recently, Ascacio-Valds et al. (2013) reported that Aspergillus niger GH1 was able to grown in solid state fermentation and produce the ellagitannase enzyme. The authors propose that the ester bonds among HHDP group and glycosides are degraded by ellagitannase which has high specicity, and this enzymatic activity allows the EA accumulation. Solid-state fermentation (SSF) consists of microbial growth and product formation on surface and inside of a porous solid matrix, in absence or near absence of free water (BarriosGonzlez, 2012). Substrate must contain enough moisture to allow microbial growth and metabolism, simulating natural growth conditions (Orza et al., 2009). Water availability in SSF is a critic limiting point which has an important inuence on microbial growth and metabolism. This availability of water corresponds to water activity (aW ), a physico-chemical parameter dened as the relative humidity of the gaseous atmosphere in equilibrium with the substrate. Chemical composition and particle structure of the supports used in SSF have a determinant inuence in the value of aW which can range from 0.80 to 0.99 to permit an efcient fungal metabolism (Martins et al., 2011). A great variety of lignocellulosic materials (LM) have been tested as solid supports for SSF, including coffee by-products (Machado et al., 2012), rice bran and wheat bran (Khandeparkar and Bhosle, 2006); sugarcane bagasse and agave (Hernndez-Salas et al., 2009; Pandey et al., 2000); mango peels (Buenrostro-Figueroa et al., 2010), grape skins (Botella et al., 2007; Rodrguez et al., 2010), cranberry pomace (Vattem and Shetty, 2003), pomegranate peels (Robledo et al., 2008); corn cobs (Mussatto et al., 2009b) and coconut husks (Orza et al., 2009) among others. Several of these by-products have been used as supports and/or substrates for production of metabolites of industrial importance, such as organic acids, antibiotics, pigments, avor and aroma compounds, bioactive molecules and a great variety of enzymes (Martins et al., 2011). The aim of this study was produce ellagitannase enzyme by growing Aspergillus niger GH1 on SSF using different agroindustrial by products as supports and partially puried ellagitannins extracted from pomegranate peels as carbon source.

candelilla (Euphorbia antisyphilitica) stalks (CS). All of them were grinded up to particle size of 0.85 mm of diameter. Before use, the matrix supports were pre-treated by boiling during 10 min and washed three times with distilled water. Support was dried at 60 C until constant weight is reached (Mussatto et al., 2009b). Physical and chemical tests consisted of water absorption index (WAI) (Orza et al., 2009), critical humidity point (CHP) (Mussatto et al., 2009a) and packing density (PD) determination (Santomaso et al., 2003). For WAI determination, the sample (1.5 g) was placed in 50 mL centrifuge tube and 15 mL of distilled water was added. The sample was stirred for 1 min at room temperature (25 C) and centrifuged at 3000 g for 10 min. The supernatant was discarded, and the WAI was calculated from the weight of the remaining gel and expressed as g gel/g dry weight. The CHP was estimated by adding 1 g of sample in a thermo-balance at 120 C for 60 min. PD was calculated by placing 10 g of sample in standard graduated cylinders and clamped to a shaker and vertically agitated until no change in volume during 5 min was observed. To be used as matrix support, the materials were pretreated by boiling during 10 min, washed three times with distilled water, and subsequently dried at 60 C for 2448 h (Mussatto et al., 2009a). Prior to use, all LM were autoclaved at 121 C for 15 min.

2.2.

Fungal strain and cell culture

Aspergillus niger GH1 strain (Food Research Department Collection, Universidad Autonoma de Coahuila, Mexico) was used. The strain has been previously isolated, characterized and identied (Cruz-Hernndez et al., 2005), highlighting their ability to degrade ellagitannins (Robledo et al., 2008; Seplveda et al., 2012). The strain was maintained at 40 C in glycerolskimmed milk. Spores of A. niger GH1 were activated in potato dextrose agar (PDA-Bioxon) medium at 30 C for ve days. The culture spores were harvested with sterile solution of 0.01% Tween-80 and counted in a Neubauer chamber.

2.3.

SSF conditions

2.

Materials and methods

2.1. Materials preparation and physico-chemical characterization

The agro industrial by-products used in this study were collected from different Mexican agricultural regions and included: sugarcane (Saccharis ofcinalis) bagasse (SB), corn (Zea mays) cobs (CC), coconut (Cocos nucifera) husk (CH) and

Ellagitannase production experiments were performed in 60 mL sterile columns (100% polypropylene) considered as bioreactor, which were aseptically packed an homogeneous mixture containing the following fermentable mass: 3 g of each support (SB, CC, CH and CS) was mixed with 7 mL of Pontecorvo culture medium (Aguilera-Carb et al., 2009) with the following composition (g L1 ): NaNO3 (6.0), KH2 PO4 (1.52), KCl (0.52), MgSO4 7H2 O (0.52), ZnSO4 (0.001), FeCl3 (0.85) and trace metals (1 mL L1 ). The trace metals solution contained (mg L1 ) Na2 B4 O7 10H2 O (10.0), MnCl2 4H2 O (50.0), Na2 MoO4 2H2 O (50.0) and CuSO4 5H2 O (250.0). Pomegranate husk ellagitannins (PHE) supplied by the Bioprocess Laboratory of the Food Research Department (School of Chemistry, Universidad Autonoma de Coahuila, Mexico) were used as carbon source and ellagitannase inducer. The medium pH was adjusted to 9 and then autoclaved (1.1 kg/m3 , 121 C) for 15 min. PHE (30 g L1 ) were added to the culture broth when the temperature was between 3540 C. Final pH was 6.5. The fermentable mass was aseptically inoculated with 2 107 spores/g of support. The SSF was carried out at 30 C for 32 h (Ascacio-Valds et al., 2013). Forced air was not supplemented for aeration of column bioreactor. Enzymatic extract (EE) was obtained by adding 7 mL of 50 mM citrates buffer pH 5 to each reactor. Fermented material was compressed and

Please cite this article in press as: Buenrostro-Figueroa, J., et al., Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation. Food Bioprod Process (2013), http://dx.doi.org/10.1016/j.fbp.2013.08.010

FBP-435; No. of Pages 7

ARTICLE IN PRESS
food and bioproducts processing x x x ( 2 0 1 3 ) xxxxxx

ltered through 0.45 lter (MilliporeTM ). Ellagitannase activity, soluble protein and biomass were determined on the EE. Volumetric enzyme productivity (g.L1 h1 ) was calculated as the ratio among the higher ellagitannase activity value and fermentation time (EAHP = EAH/t).

Table 1 Water absorption index (WAI), critical humidity point (CHP) and packing density (PD) of different agro-industrial wastes*. Support
Coconut husk (CH) Corn cobs (CC) Sugarcane bagasse (SB) Candelilla stalks (CS)

WAI (g/g dw)

12.09a 2.97cd 9.46ab 3.14c

CHP (%)
16b 27c 12a 29.5cd

PD (g/cm3 )
0.82b 0.74a 0.83b 0.86c

2.4.

Analytical procedures

Protein content was analyzed using bovine serum albumin solution at 100 ppm (10 mg in 100 mL of 50 mM citrate buffer pH 5). For assay 100 L of sample was added with 1000 L of Bradford reagent. The samples were shaken and allowed to rest ve minutes. Absorbance was recorded at 595 nm (Bradford, 1976). Fungal biomass was assayed following glucosamine method reported by Blix (1948). Samples were hydrolyzed in order to release the glucosamine from cell wall, the pyrrole compound formed when combined with acetylacetone reacts with p-dimethylaminobenzaldehyde forming a red compound detected at 530 nm. A calibration curve (0200 mg.mL1 of glucosamine) was carried out at the same experimental conditions than the samples. Glucosamine associated with fungal growth was determined to obtain the biomass content (mg g1 of sample). Ellagitannase activity was assayed according to (AscacioValds et al., 2013). Pomegranate husk ellagitannins [PEH (1 mg.mL1 ) in 50 mM citrate buffer pH 5] were used as enzyme substrate. PEH contain punicalagin and punicalin, both of them are ellagitannins molecules that releases ellagic acid when are subjected to hydrolysis. An enzyme treatment control (1000 L ECG + 50 L 50 mM citrate buffer pH 5), an extract treatment control (1000 L of 50 mM citrate buffer pH 5 + 50 L of enzymatic extract) and the reaction mixture (1000 L PEH + 50 L to enzymatic extract) were prepared. All enzymatic preparations were allowed to react for 10 min at 60 C in a water bath (Sheldon manufacturing m. 1225). The reaction was terminated by adding 1050 L of absolute ethanol. Then samples were sonicated for 25 min, ltered through 0.45 m membrane units (Millex ) and collected in vials. Ellagic acid quantication was carried out by HPLC (High Performance Liquid Chromatography) equipment (Varian ProStar System) with a Diode Array Detector (PDA ProStar) to 254 nm, according to Ascacio-Valds et al. (2010), under the following operation conditions: 5 m Optisil ODS column (250 4.6 mm), ow rate of 1 mL min1 , sample volume of 10 L, 30 C in column for 40 min, with acetonitrile and 3% acetic acid as mobile phase. Ellagic acid (0500 ppm) stock solution was prepared for calibration curve. One ellagitannase enzymatic unit was dened as the enzyme amount needed to release 1 mol of ellagic acid per min under the above conditions.

*There are no signicant differences among the same letters.

were sugarcane bagasse (SB), coconut husk (CH), corn cobs (CC) and candelilla stalks (CS). Biomass production, protein soluble, ellagitannase, volumetric and specic productivities were determinated. All treatments were realized by three replications. Data were analyzed by ANOVA using SAS 9.0, when needed mean treatments were compared using Tukeys multiple range procedure. A p-value of less than 0.05 was regarded as signicantly different.

3.
3.1.

Results and discussion

Physico-chemical characterization

2.5.

Scanning electron microscopy

Visualization of spore and fungal growth was done using a Phillips XL30-ESEM (Environmental Scanning Electron Microscope). Samples dehydrated were coated with electrolytic gold (99.99% purity) in a vacuum evaporator Jeol Model JEE-400. All samples were analyzed under vacuum with a GSE (Gaseous Secondary Electron) detector.

2.6.

Experimental design and data analysis

The effect of support on ellagitannase production was evaluated under a completely randomized design. The treatments

The SSF process requires the use of low water content materials to facilitate fungal growth and development, due to the type of support plays an important role in yielding the higher growth rates of microorganisms (Manpreet et al., 2005). WAI and CHP are parameters highly relevant when evaluating the potential of different materials used as support in SSF (Mussatto et al., 2009b; Orza et al., 2009). WAI indicates sample capacity to absorb water, depending on the availability of hydrophilic groups which bind water molecules, and on the gel forming capacity of macromolecules (Mussatto et al., 2009a). The highest WAI value was found in CH, which was four times higher than those obtained for CC and CS. SB was three times higher than CC and CS. No signicant differences (p 0.05) were observed between CH and SB values (Table 1). According to (Robledo et al., 2008), materials with high WAI are preferred since facilitate microorganism growth and development. For CC, WAI are similar to those reported by Orza et al. (2009). For CH, WAI are similar to those reported by Mussatto et al. (2009b). CHP represents the amount of water linked to the support, which cannot be used by the microorganism for their metabolic functions. The materials must have low CHP to facilitate microorganism culture. High values of CHP can affect microorganism growth because a high proportion of water is bonded to the support material, and consequently, microbial species development will be affected, due to free water is not much (Martins et al., 2011). Moo-Young et al. (1983) recommended a maximum limit of CHP at 40% for A. niger strains in SSF, due to the need for modication of the moisture content in relation to the absorbed media. Table 1 shows the CHP values obtained in the present work for each agroindustrial by-product assay. All the supports tested have CHP values under 40%, limit recognized for the growth of A. niger in SSF (Moo-Young et al., 1983), however those obtained in CH and SB are up two times lower than those obtained in CC and CS. CHP are lower than those previouslly reported for CC and CH (Mussatto et al., 2009b; Orza et al., 2009). The high WAI and low CHP values found in CH and SB make it an excellent

Please cite this article in press as: Buenrostro-Figueroa, J., et al., Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation. Food Bioprod Process (2013), http://dx.doi.org/10.1016/j.fbp.2013.08.010

FBP-435; No. of Pages 7

ARTICLE IN PRESS
food and bioproducts processing x x x ( 2 0 1 3 ) xxxxxx

Fig. 1 Kinetics of: (a) biomass production, (b) ellagitannase activity, (c) volumetric productivity and (d) specic productivity obtained by SSF with A. niger GH1 using as supports coconut husk (CH), candelilla stalks (CS), sugarcane bagasse (SB) and corn cobs (CC).

materials to be used as a support on SSF, compared with CC and CS. Packing density (PD) is another important parameter to evaluate for the SSF processes, due to provides the material compaction degree, therefore, the available space for mass and energy transfer (Chvez-Gonzlez et al., 2010). The lowest PD value (Table 1) was obtained with CC (0.74 g cm3 ). There are no difference in PD values among CH and SB. A high value of PD was found in CS, which might to produce problems in mass and energy transference.

3.2.

Ellagitannase production in SSF

Supports can also provide carbon and nutriment during the fermentation process, or as bonding surface for fungus invasion in the impregnated support with culture medium. Fungi are microorganisms easily adaptable to SSF since their hyphae can grow over the particles surface and even penetrate the intra-individual spaces and colonize the solid support substrate (Graminha et al., 2008). A. niger GH1 was able to grow invading and penetrating the different supports, associated with ellagitannins biodegradation by the ellagitannase enzyme. In this work, A. niger GH1 presents a fast biomass production reaching the maximum value (400 mg g1 ) in SB, this value is two-fold better than those obtained with CH, CS and CC (Fig. 1(a)). This difference may be attributed to presence of other compounds in the SB after the wash process, such as polysaccharides and monomeric sugars, which are prefer to the fungus growth. The support-substrate invasion was associated to ellagitannase activity (Fig. 1(b)). It was observed that ellagitannase

activity appeared after 8 h of culture, reached the maximum enzyme activity among 16 and 24 h for all tested supports, after this the enzymatic activity decreases, maybe attributed to production of other enzymes, such as proteases, which may generate enzyme degradation during the ellagitannins degradation process. Maximum ellagitannase activities were found in SB and CC (1404.01 U L1 and 1179.98 U L1 respectively), higher than CH (925 U L1 ) and 2.5 times more than that obtained using CS (650 U L1 ). No signicant differences between CC and SB at 24 h (p 0.05) were observed. The lowest value of ellagitannase activity was found in CS, may be due to the high CHP value which limits availability of free water for fungus growth and metabolism. On the other hand, PD value found in CS was higher than those observed for all others supports (Table 1). An increase in packing density may cause a reduction in the void space between particles and a concomitant reduction in the area of exchanged with the surrounding atmosphere (Barrios-Gonzlez et al., 1993). With CH was observed the highest enzyme volumetric productivity (115.62 U L h1 ), two more times than CC and SB, and up to four times higher than CS (Fig. 1(c)). The use of puried pomegranate ellagitannins as ellagitannase inducers in SSF with A. niger GH1 fungus allowed to obtain high enzymatic titers and good specic productivity. The best specic productivity was obtained using CC, being two more times than SB and CH at the same fermentation time (Fig. 1(d)). Environmental scanning electron microscopy demonstrated that A. niger GH1 grew invading and penetrating the corn cobs support (Fig. 2). Pinto et al. (2012) reported that corn cob is composed by three different layers, clearly different among them. Corn cobs present a porous particle layer

Please cite this article in press as: Buenrostro-Figueroa, J., et al., Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation. Food Bioprod Process (2013), http://dx.doi.org/10.1016/j.fbp.2013.08.010

FBP-435; No. of Pages 7

ARTICLE IN PRESS
food and bioproducts processing x x x ( 2 0 1 3 ) xxxxxx

Fig. 2 Scanning electronic microscope images of the fungus Aspergillus niger GH1 growth on corn cobs particles. (A) Without fungus; (B), (C) and (D): with fungus. (Fig. 2(A)). After fermentation, mycelial growth was observed on and inside corn cob cavities (Fig. 2(B)). Furthermore, it was observed other microstructure with regular geometric shape and regular alveolar forms (Fig. 2(C)). In this gure, it was observed spores of A. niger GH1, which possess a rough structure surface (Fig. 2(D)). This support has been reported to promote high enzymatic production values. Evaluating fructooligosaccharides and -fructofuranosidase production by A. japonicus on different lignocellulosic materials as support, Mussatto et al. (2009b) achieved the highest productivity values using CC under submerged fermentation (SmF). There are a few reports about biodegradation of pomegranate ellagitannins by microorganisms, however in recent times, important works have been developed. Ellagitannase activity and productivities values obtained in this study are 31 and up to 140 more times than those reported by Aguilera-Carb (2009) whom used SSF of pomegranate husks and the same A. niger GH1 strain. This could be explained by the fact that they used a complex substrate which consists in compounds such as polysaccharides, proteins, monomeric sugars and of course, polyphenols, where the fungus metabolized other molecules such as sugars leaving for later time polyphenols and therefore the enzymes responsible for the degradation of the latter will be expressed in different time of culture. Previously, ellagic acid production has been reported by Aspergillus oryzae and Trichoderma reesei in SmF, using as substrate and carbon source valonea extracts (Huang et al., 2007), whom obtained 300 U L1 ellagitannin acyl hydrolase activity, this value was lower than that obtained with candelilla stalks in this study and up to 4.7 times lower than that achieved with CC and SB. Due to the obtained results, a selection criterion (S) was established to choose the best support. The response variables [ellagitannase activity (ea), specic activity (sa), volumetric productivity (vp) and specic productivity (sp)] were weighted according to their importance in the process (S = [1.5 (ea) + 2.0 (sa) + 2.5 (vp) + 4.0 (sp)]). The highest value was given to the specic productivity (U mg protein h1 ) as an indicator of process efciency and purity of the enzyme obtained. All supports were suitable for use in ellagitannase production by SSF with A. niger GH1 (Fig. 3), excepts candelilla stalks. No signicant differences between SB, CH and CC were found. In SSF processes using impregnated supports with culture medium is important to take into account aspects such as cost and

Fig. 3 Selection index (-) for use of sugarcane bagasse (SB), coconut husk (CH), corn cobs (CC) and candelilla stalks (CS) as supports in ellagitannase production by SSF.

Please cite this article in press as: Buenrostro-Figueroa, J., et al., Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation. Food Bioprod Process (2013), http://dx.doi.org/10.1016/j.fbp.2013.08.010

FBP-435; No. of Pages 7

ARTICLE IN PRESS
food and bioproducts processing x x x ( 2 0 1 3 ) xxxxxx

availability of support, environmental impact of the solids produced, as wells as process cost and product. The use of SB, CH and CC as a matrix for the culture media absorption and attachment matrix for the fungal growth provides important advantages, such as higher production titers and productivity. However, the use of CC may signicantly decrease costs production, due to high specic productivity at short times compared with the other materials evaluated, facilitates the recovery of interest compounds, and can be reused, eliminating the needed time for fungus growth. The next step will be evaluated these fermented materials in the ellagitannins hydrolysis for continuous ellagic acid production.

4.

Conclusions

According to the obtained results from the physico-chemical analysis (WAI, CHP and PD), all tested agro-industrial byproducts (sugarcane bagasse, coconut husk, corn cobs and candelilla stalks) have great potential to be used as support in SSF process. A. niger GH1 secreted in higher amounts the ellagitannase enzyme, in order to break the ester bonds among HHDP group and glycosides to release ellagic acid. This suggests the induction of ellagitannase by partially puried ellagitannins. Corn cobs, sugarcane bagasse and coconut husk can be used as excellent support in SSF for the ellagitannase production with high titers. However the use of corn cobs provides the highest specic productivity at shorter times, which may decreases the production costs. The development of a bioprocess for ellagitannase production would offer economic and environmental advantages in ellagic acid production compared with chemical methods.

Acknowledgement
Juan Buenrostro wants to thanks the Mexican Council of Science and Technology (CONACyT) for the scholarship assigned for their postgraduate studies in the program of Food Science and Technology, UAdeC.

References
Aguilera-Carb, A., Augur, C., Prado-Barragn, L., Aguilar, C., Favela-Torres, E., 2008a. Extraction and analysis of ellagic acid from novel complex sources. Chem. Pap. 62, 440444. Aguilera-Carb, A., Augur, C., Prado-Barragn, L., Favela-Torres, E., Aguilar, C., 2008b. Microbial production of ellagic acid and biodegradation of ellagitannins. Appl. Microbiol. Biotechnol. 78, 189199. Aguilera-Carb, A., Hernndez, J., Augur, C., Prado-Barragn, L., Favela-Torres, E., Aguilar, C., 2009. Ellagic acid production from biodegradation of Creosote Bush ellagitannins by Aspergillus niger in solid state culture. Food Bioprocess Tech. 2, 208212. Aguilera-Carb, F.A., 2009. Ellagic Acid Production: Enzymatics Studies. Universidad Autnoma Metropolitana Iztapalapa, Mxico, D.F., pp. 131. Ascacio-Valds, J., Aguilera-Carb, A., Martnez-Hernndez, J., Rodrguez-Herrera, R., Aguilar, C., 2010. Euphorbia antisyphilitica residues as a new source of ellagic acid. Chem. Pap. 64, 528532. Ascacio-Valds, J.A., Buenrostro-Figueroa, J.J., Aguilera-Carb, A., Prado-Barragn, A., Rodrguez-Herrera, R., Aguilar, C.N., 2011. Ellagitannins: biosynthesis, biodegradation and biological properties. J. Med. Plants Res. 5, 46964703.

Ascacio-Valds, J.A., Buenrostro, J.J., De la Cruz, R., Seplveda, L., Aguilera, A.F., Prado, A., Contreras, J.C., Rodrguez, R., Aguilar, C.N., 2013. Fungal biodegradation of pomegranate ellagitannins. J. Basic Microbiol. 0, 17. Barrios-Gonzlez, J., 2012. Solid-state fermentation: physiology of solid medium, its molecular basis and applications. Process Biochem. 47, 175185. Barrios-Gonzlez, J., Gonzlez, H., Meja, A., 1993. Effect of particle size, packing density and agitation on penicillin production in solid state fermentation. Biotechnol. Adv. 11, 539547. Blix, G., 1948. The determination of hexosamines according to elson and morgan. Acta Chem. Scand. 2, 467473. Botella, C., Diaz, A., de Ory, I., Webb, C., Blandino, A., 2007. Xylanase and pectinase production by Aspergillus awamori on grape pomace in solid state fermentation. Process Biochem. 42, 98101. Bradford, M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248254. Buenrostro-Figueroa, J., de la Garza-Toledo, H., Ibarra-Junquera, V., Aguilar, C., 2010. Juice extraction from mango pulp using an enzymatic complex of Trichoderma sp. produced by solid-state fermentation. Food Sci. and Biotechnol. 19, 13871390. Cruz-Hernndez, M., Contreras-Esquivel, J., Lara, F., Rodrguez-Jasso, R., Aguilar, C., 2005. Isolation and evaluation of tannin-degrading fungal strains from the Mexican desert. Z. Naturforsch. 60, 844848. Chvez-Gonzlez, M.L., Rodrguez-Durn, L.V., Cruz-Hernndez, M., Prado-Barragn, A., Aguilar, C.N., 2010. Packing density and aeration rate effect on tannase production by Aspergillus niger GH1 in solid state fermentation. In: Sabu, A., Aguilar, C.N., Roussos, S. (Eds.), Chemistry and Biotechnology of Polyphenols: Fundamentals and Application. CiBET, Kerala, India, p. 199. Graminha, E.B.N., Gonc alves, A.Z.L., Pirota, R.D.P.B., Balsalobre, M.A.A., Da Silva, R., Gomes, E., 2008. Enzyme production by solid-state fermentation: application to animal nutrition. Anim. Feed Sci. Technol. 144, 122. Gross, G.G., 2009. Biosynthesis of ellagitannins: old ideas and new solutions. In: Quideau, S. (Ed.), Chemistry and Biology of Ellagitannins: An Underestimated Class of Bioactive Plant Polyphenols. World Scientic Publishing, Singapore, pp. 94118. Hernndez-Salas, J.M., Villa-Ramrez, M.S., Veloz-Rendn, J.S., Rivera-Hernndez, K.N., Gonzlez-Csar, R.A., Plascencia-Espinosa, M.A., Trejo-Estrada, S.R., 2009. Comparative hydrolysis and fermentation of sugarcane and agave bagasse. Bioresour. Technol. 100, 12381245. Huang, W., Ni, J., Borthwick, A.G.L., 2005. Biosynthesis of valonia tannin hydrolase and hydrolysis of valonia tannin to ellagic acid by Aspergillus SHL 6. Process Biochem. 40, 12451249. Huang, W., Niu, H., Li, Z., Lin, W., Gong, G., Wang, W., 2007. Effect of ellagitannin acyl hydrolase, xylanase and cellulase on ellagic acid production from cups extract of valonia acorns. Process Biochem. 42, 12911295. Khadem, S., Marles, R., 2010. Monocyclic phenolic acids; hydroxyand polyhydroxybenzoic acids: occurrence and recent bioactivity studies. Molecules 15, 79858005. Khandeparkar, R.D.S., Bhosle, N.B., 2006. Isolation, purication and characterization of the xylanase produced by Arthrobacter sp. MTCC 5214 when grown in solid-state fermentation. Enzyme Microb. Technol. 39, 732742. Koponen, J.M., Happonen, A.M., Mattila, P.H., Trrnen, A.R., 2007. Contents of anthocyanins and ellagitannins in selected foods consumed in Finland. J. Agric. Food Chem. 55, 16121619. Machado, E.M.S., Rodrguez-Jasso, R.M., Teixeira, J.A., Mussatto, S.I., 2012. Growth of fungal strains on coffee industry residues with removal of polyphenolic compounds. Biochem. Eng. J. 60, 8790. Manpreet, S., Sawraj, S., Sachin, D., Pankag, S., Banerjee, U.C., 2005. Inuence of process parameters on the production of metabolites in solid-state fermentation Malalaysian. J. Microbiol. 1, 19.

Please cite this article in press as: Buenrostro-Figueroa, J., et al., Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation. Food Bioprod Process (2013), http://dx.doi.org/10.1016/j.fbp.2013.08.010

FBP-435; No. of Pages 7

ARTICLE IN PRESS
food and bioproducts processing x x x ( 2 0 1 3 ) xxxxxx

Martins, S., Mussatto, S.I., Martnez-Avila, G., Montanez-Saenz, J., Aguilar, C.N., Teixeira, J.A., 2011. Bioactive phenolic compounds: production and extraction by solid-state fermentation. A review. Biotechnol. Adv. 29, 365373. Moo-Young, M., Moreira, A., Tengerdy, R., 1983. Principles of Solid Substrate Fermentation. In: Smith, J., Berry, D., Kristiansen, B. (Eds.), The Filamentous Fungi. Edward Arnold Publishers, London, pp. 117144. Mussatto, S.I., Aguilar, C.N., Rodrigues, L.R., Teixeira, J.A., 2009a. Colonization of Aspergillus japonicus on synthetic materials and application to the production of fructooligosaccharides. Carbhydr. Res. 344, 795800. Mussatto, S.I., Aguilar, C.N., Rodrigues, L.R., Teixeira, J.A., 2009b. Fructooligosaccharides and -fructofuranosidase production by Aspergillus japonicus immobilized on lignocellulosic materials. J. Mol. Catal. B: Enzym. 59, 7681. Orza, M.C., Mussatto, S.I., Contreras-Esquivel, J.C., Rodrguez, R., de la Garza, H., Teixeira, J.A., Aguilar, C.N., 2009. Exploitation of agro industrial wastes as immobilization carrier for solid-state fermentation. Ind. Crop. Prod. 30, 2427. Pandey, A., Soccol, C.R., Nigam, P., Soccol, V.T., 2000. Biotechnological potential of agro-industrial residues. I: sugarcane bagasse. Bioresour. Technol. 74, 6980. Pinto, J., Cruz, D., Paiva, A., Pereira, S., Tavares, P., Fernandes, L., Varum, H., 2012. Characterization of corn cob as a possible raw building material. Constr. Build. Mater. 34, 2833. Robledo, A., Aguilera-Carb, A., Rodrguez, R., Martnez, J., Garza, Y., Aguilar, C., 2008. Ellagic acid production by Aspergillus niger in solid state fermentation of pomegranate residues. J. Ind. Microbiol. Biotechnol. 35, 507513.

Rodrguez, L.A., Toro, M.E., Vazquez, F., Correa-Daneri, M.L., Gouiric, S.C., Vallejo, M.D., 2010. Bioethanol production from grape and sugar beet pomaces by solid-state fermentation. Int. J. Hydrogen Energy 35, 59145917. Santomaso, A., Lazzaro, P., Canu, P., 2003. Powder owability and density ratios: the impact of granules packing. Chem. Eng. Sci. 58, 28572874. Seeram, N., Lee, R., Hardy, M., Heber, D., 2005. Rapid large scale purication of ellagitannins from pomegranate husk, a by-product of the commercial juice industry. Sep. Purif. Technol. 41, 4955. Seplveda, L., Aguilera-Carb, A., Ascacio-Valds, J.A., Rodrguez-Herrera, R., Martnez-Hernndez, J.L., Aguilar, C.N., 2012. Optimization of ellagic acid accumulation by Aspergillus niger GH1 in solid state culture using pomegranate shell powder as a support. Process Biochem. 47, 21992203. Shi, B., Qiang, H., Kai, Y., Wen, H., Qing, L., 2005. Production of ellagic acid from degradation of valonea tannins by Aspergillus niger and Candida utililis. J. Chem. Technol. Biotechnol. 80, 11541159. Vattem, D.A., Shetty, K., 2002. Solid-state production of phenolic antioxidants from cranberry pomace by Rhizopus oligosporum. Food Biotechnol. 16, 189210. Vattem, D.A., Shetty, K., 2003. Ellagic acid production and phenolic antioxidant activity in cranberry pomace (Vaccinium macrocarpon) mediated by Lentinus edodes using a solid-state system. Process Biochem. 39, 367379. Wilson, T.C., Hagerman, A.E., 1990. Quantitative determination of ellagic acid. J. Agr. Food Chem. 38, 16781683.

Please cite this article in press as: Buenrostro-Figueroa, J., et al., Potential use of different agroindustrial by-products as supports for fungal ellagitannase production under solid-state fermentation. Food Bioprod Process (2013), http://dx.doi.org/10.1016/j.fbp.2013.08.010