PROTEASES FROM CELL CULTURE OF Bromelia hemisphaerica ...

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FSB1 – 2004 Food Science and Biotechnology in Developing Countries activity of 37640U/L*min in 72 hours. These results show that addition of an enzymatic repressor at low levels have positive effect in enzyme production, the activity up 17.34 times with relation at only inductor production. REFERENCES Aguilar, C.N. 2000. Inducción y represión de la síntesis de la tanasa de Aspergillus níger Aa-20 en cultivo en medios líquido y sólido. Tesis de doctorado. Universidad Autónoma Metropolitana. Unidad Iztapalapa. México, D.F. Aguilar, C.N., Augur, C., Favela-Torres, E. and Viniegra-González, G. 2001. Induction and repression patterns of fungal tannase in solid state and submerged cultures. Process Biochemistry. 36(6), 571-578. Ashokkumar, B., Kayalvizhi, N. and Gunasekaran, P. 2001. Optimization of media for βfructofuranosidase production by Aspergillus niger in submerged and solid state fermentation. Process Biochemistry. 37, 331-338. Miller, G.L. 1959. Use of Dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry. 31, 426-428. Romero-Gomez, S.J., Augur, C. and Viniegra-Gonzalez, G. 2000. Invertase production by Aspergillus niger in submerged and solid state fermentation. Biotechnology Letters. 22, 1255- 1258.
FSB1 – 2004 Food Science and Biotechnology in Developing Countries PRODUCTION, PARTIAL PURIFICATION AND CHARACTERIZATION <strong>OF</strong> A FUNGAL TANNASE PRODUCED BY SOLID STATE FERMENTATION. Mario Cruz-Hernández, Juan Carlos Contreras-Esquivel, Raúl Rodríguez-Herrera and Cristóbal Noé Aguilar* Food Research Dept. School of Chemistry. Universidad Autónoma de Coahuila. Saltillo, Coahuila, México. *Corresponding author: cag13761@mail.uadec.mx ABSTRACT Production and partial characterization of a fungal tannase was studied. Aspergillus niger GH1 was selected by its capacity for tannase production in the solid state culture (SSC). Several culture conditions were evaluated. A protocol for tannase pre-purification was developed. It included concentration by polyethilenglycol (PEG), ionic exchange chromatography (IEC) and isoelectric focusing Rotofor ® . Two tannase isoforms were found in the culture broth. Key words: tannase, production, partial purification, partial characterization, solid state fermentation. INTRODUCTION Tannase or tannin acyl hydrolase (EC, 3.1.1.20) catalyzes the hydrolysis reaction of the ester bonds present in the hydrolysable tannins and gallic acid esters. Its production at industrial level is by microbial way using SmC, where the activity is expressed mainly of intracellular form, implying additional costs in its production (Lekha and Lonsane, 1994). At the moment, the main commercial applications of the tannase are given in the elaboration of instantaneous tea or of acorn liquor and in the production of the gallic acid (Coggon et al., 1975; Chae and Yu, 1983; Pourrat et al., 1985; García- Najera et al., 2002), which is an important intermediary compound in the synthesis of the antibacterial drug, trimetroprim, used in the pharmaceutical industry (Sittig, 1988) and also in the food industry; gallic acid is a substrate for the chemical or enzymatic synthesis of the propylgallate, a potent antioxidant. Also, the tannase is used as clarifying agent in some wines, juices of fruits and in refreshing drinks with flavour to coffee (Lekha et al., 1993). Several studies have reported interesting advantages between the tannase produced by SSC in relation with that produced by SmC. On this topic there are few reports, but they are clearly interesting (Chaterjee et al., 1996; Lekha and Lonsane 1997; García-Peña et al., 1999; Ramírez-Coronel et al., 1999; Aguilar et al., 1999; Aguilar et al., 2001a, 2001b and 2002a; Van de Lagemaat and Pyle, 2001). In these, attractive advantages indicated, are the high production titles (up to 5.5 times more than in SmC), the nature extracellular of the enzymes and the stability to wide pH and temperature ranges (Lekha and Lonsane, 1994). This study was aimed to evaluate several culture conditions for production partial purification and characterization of fungal tannase MATERIAL AND METHODS Microorganism The strain of Aspergillus niger GH1 (DIA/UAdeC-Collection) was selected after several comparative studies between tannin-degrading strains (Cruz-Hernández et al., 2002). Spores were maintained at - 20ºC under a crio-protector system.
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PROTEASES FROM CELL CULTURE OF Bromelia hemisphaerica ...

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