Indian Journal of Science and Technology

Open Access Open Access  Restricted Access Subscription Access

Optimization of Extracellular Lipase Production in Colletotrichum Gloeosporioides by Solid State Fermentation


Affiliations
1 Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai-600 025, Tamil Nadu, India
 

Twenty two endophytic fungi isolated from oil seeds were screened for their ability to produce extracellular lipolytic enzymes in solid state fermentation. Among them, the most productive strain, identified as Colletotrichum gloeosporioides was cultivated for lipase production in solid state fermentation (SSF) using oil-mill residue as a solid support. The elaboration of extracellular lipases from C. gloeosporioides on different residual cheap oil substrates was tested. Among these, pongamia oil cake (POC) showed maximum lipase activity (983 U/g dry matter (DM)) followed by coconut oil cake (COC) (925 U/g DM). These two oil industrial residues were chosen for enhanced lipase production. The maximum enzyme yield was obtained when POC was impregnated with CDB (Czapek-Dox Broth) in the ratio of 1:1.5 and with (906 U/g DM). Triton X 100 as the extractant from the fermented solids yielded 1020 U/g DM. The optimum temperature for lipase secretion by C. gloeosporioides was found to be 25°C. Xylose as the carbon source and peptone as the nitrogen source were favorable for the secretion of the enzymes. Tween 60 served as a good lipid substrate for lipase production (1170U/g DM) when POC was the substrate. Amending the POC with Magnesium sulphate as a metallic ion source induced good lipase activity (1240 U/g DM) by C. gloeosporioides. The Sunflower oil was found to be best co-vegetable oil substrate to induce the enzyme (2560 U/g DM). Triton X100 served as the surfactant for lipase secretion in C. gloeosporioides. Thus, pongamia oil cake can be used as cheap substrate for enhanced extracellular lipase production by C. gloeosporioides.

Keywords

Lipases, Colletotrichum gloeosporioides, Solid State Fermentation, Pongamia Oil Cake, Coconut Oil Cake
User

  • Al- Asheh S and Duvnjak Z (1994) The effect of surfactants on the phytase production and the reduction of the phytic acid in canola meal by Aspergillus carbonarius using a solid state fermentation process. Biotechnol. Lett. 2, 183- 188.

  • Benjamin S and Pandey A (1997) Coconut cake- a potent substrate for the production of lipase by Candida rugosa in solid-state fermentation. Acta Biotechnol. 17, 241-251.

  • Benjamin S and Pandey A (1998) Mixed-solid substrate fermentation. A novel process for enhanced lipase production by Candida rugosa. Acta Biotechnol. 18, 315-324.

  • Bogar B, Szak´acs G, Pandey A, Abdulhameed S, Linden JC and Tengerdy RP (2003) Production of phytase by Mucor racemosus in solid-state fermentation. Biotechnol. Prog. 19, 312-319.

  • Cammarota MC and Freire DMG (2006) A review on hydrolytic enzymes in the treatment of wastewater with high oil and grease content. Bioresour. Technol. 97, 2195–2210.

  • Castilho LR, Polato CMS, Baruque EA, Sant’Anna jr GL and Freire DMG (2000) Economic analysis of lipase production by Penicillium restrictum in solid-state and submerged fermentations. Biochem. Eng. J. 4,239–247.

  • Colen G, Junqueira RG and Moraes- Santos T (2006) Isolation and Screening of alkaline lipase- producing fungi from Brazilian Savanna soil. World J. Microbiol. Biotechnol. 22, 881- 885.

  • Cordova J, Nemmaoui M, Ismaili-Alaoui M, Morin A, Roussos S, Raimbault M and Benjilali B (1998) Lipase production by solid state fermentation of olive cake and sugar cane bagasse. J. Mol. Catal. B: Enzym. 5, 75-78.

  • Di Luccio M, Capra F, Ribeiro NP, Vargas GDLP, Freire DMG, Denise MG and De Oliveira D (2004) Effect of temperature, moisture, and carbon supplementation on lipase production by solid-state fermentation of soy cake by Penicillium simplicissimum. Appl. Biochem. Biotechnol. 113– 116, 173-180.

  • Dominguez A, Costas M, Longo MA and Sanroman A (2003) A novel application of solid state culture: Production of lipases by Yarrowia lipolytica. Biotechnol. Lett. 25, 1225-1229.

  • Durand A (2003) Bioreactor designs for solid state fermentation. Biochem. Eng. J. 13,113- 125.

  • Ebune A, Al- Asheh S and Duvnjak Z (1995) Effect of phosphates, surfactants and glucose on phytase production and hydrolysis of phytic acid in canola meal by Aspergillus ficcum during solid state fermentation. Bioresour. Technol . 54, 241-247.

  • Freire DMG, Teles EMF, Bon EPS and Sant Anna GL (1997) Lipase production by Penicillium restrictum in a bench-scale fermenter effect of carbon and nitrogen nutrition agitation and aeration. Appl. Biochem. Biotechnol. 63/64, 409-421.

  • Gombert AK, Pinto AL, Castilho LR and Freire DMG (1999) Lipase production by Penicillium restrictum in solid-state fermentation using babassu oil cake as substrate. Proc. Biochem. 35, 85-90.

  • Gutarra MLE, Godoy MG, Castilho LR, Freire DMG (2007) Inoculum strategies for Penicillium simplicissimum lipase production by solid-state fermentation using a residue from the babassu oil industry. J. Chem. Technol. Biotechnol. 82, 313–318.

  • H¨olker U, H¨ofer M and Lenz J (2004) Biotechnological advantages of laboratoryscale solid-state fermentation with fungi. Appl. Microbial. Biotechnol. 64, 175–186.

  • Kamini NR, Mala JGS, Puvanakrishnan P (1998) Lipase production from Aspergillus niger by solid-state fermentation using gingelly oil cake. Proc. Biochem. 33, 505-511.

  • Kwon DY and Rhee JS (1986) A simple and rapid colorimetric method for determination of free fatty acids for lipase assay. JAOCS , 63 (1), 89- 92.

  • Leal MCMR, Cammarota MC, Freire DMG and Sant’Anna GL (2002) Hydrolytic enzymes as coadjutants in the anaerobic treatment of dairy wastewaters. Brazil. J. Chem. Eng.19, 175- 180.

  • Macedo GA, Lozano MMS and Pastore GM (2003) Enzymatic synthesis of short chain citronellyl esters by a new lipase from Rhizopus sp. Electr. J. Biotechnol. 6, (http://www.ejbiotechnology.info/content/ vol6/issue1/full/2/2.pdf).

  • Mahadik ND, Puntambekar US, Bastawde KB, Khire JM and Gokhale DV (2002) Production of acidic lipase by Aspergillus niger in solid state fermentation. Proc. Biochem. 38 (5), 715-721.

  • Olama ZA and El-Sabaeny AH (1993) Lipase production by Aspergillus niger under various growth conditions using solid state fermentation. Microbiologia (Madrid) 9, 134- 141.

  • Ortiz-Vazquez E, Granados-Baeza M and Rivera-Munoz G (1993) Effect of culture conditions on lipolytic enzyme production by Penicillium candidum in a solid state fermentation. Biotechnol. Adv. 11, 409-416.

  • Palma MB, Pinto AL, Gombert AK, Seitz KH, Kivatinitz SC, Castilho LR and Freire DMG (2000) Lipase production by Penicillium restrictum using solid waste of industrial Babassu oil production as substrate. Appl. Biochem. Biotechnol. 84–86, 1137- 1146.

  • Pandey A, Benjamin S, Soccol CR, Nigam P, Krieger N and Soccol VT (1999a) The realm of microbial lipases in biotechnology. Biotechnol. Appl. Biochem. 29, 119-131.

  • Pandey A, Selvakumar P, Soccol CR and Nigam P (1999b) Solid state fermentation for the production of industrial enzymes. Curr. Sci. 77, 149-162.

  • Pandey A, Soccol CR and Mitchell D (2000) New developments in solid state fermentation: 1-bioprocesses and products. Proc. Biochem. 35, 1153-1169.

  • Raimbault M and Alazard D (1980) Culture method to study fungal growth in solid fermentation. Eur. J. Appl. Microbiol. Biotechnol. 9, 199-209.

  • Ramachandran S, Patel AK, Nampoothiri KM, Franics F, Nagy V and Szakacs G (2004) Coconut oil cake – a potential raw material for the production of a – amylase. Bioresour. Technol, 93, 169-174.

  • Rao PV, Jayaraman K and Lakshmanan CM (1993a) Production of lipase by Candida rugosa in solid state fermentation. 1: determination of significant process variables. Proc. Biochem. 28, 385-389.

  • Rao PV, Jayaraman K and Lakshmanan CM (1993b) Production of lipase by Candida rugosa in solid state fermentation. 2. Medium optimization and effect of aeration Proc. Biochem. 28, 391-395.

  • Rivera-Munoz G, Tinoco-Valencia JR, Sanchez S and Farres A (1991) Production of microbial lipases in a solid state fermentation system. Biotechnol. Lett. 13, 277-280.

  • Rodriguez JA, Mateos JC, Nungaray J, Gongalez V, Bhagnagar T, Roussos S, Cordova J and Baratti J (2006) Improving lipase production by nutrient source modification using Rhizopus homothallicus cultured in solid state fermentation. Proc. Biochem. 41, 2264-2269.

  • Sharma R, Chisti Y and Banerjee UC (2001) Production, purification, characterization, and applications of lipases. Biotechnol.19, 627–662.

  • Sugiura M (1984) Bacterial lipases. In: Borgstrom B, Brockman H.L, editors. Lipases. Amsterdam: Elsevier. 505-523.

  • Sumitra R, Singh SK, Larroche C, Soccol CR and Pandey A (2007) Oil cakes and their biotechnological applications- A review. Bioresour. Technol. 98 (10), 2000- 2009.

  • Tan T, Zhang M, Wang B, Ying C and Deng L (2003) Screening of high lipase production Candida sp. and production of lipase by fermentation. Proc. Biochem. 39, 459-465.

  • Tan T, Zhang M, Xu J and Zhang J (2004) Optimization of culture conditions and properties of lipase from Penicillium camembertii Thom PG-3. Proc. Biochem. 39,1495-1502.

  • Ul-Haq I, Idrees S and Rajoka MI (2002) Production of lipases by Rhizopus oligosporous by solid-state fermentation. Proc. Biochem. 37, 637-641.

  • Uvarani G, Jaganathan L, Shridas P and Boopathy R (1998) Purification and characterization of lipase from Rhizomucor miehei. J. Sci. Ind. Res. 57, 607-610.


Abstract Views: 564

PDF Views: 527




  • Optimization of Extracellular Lipase Production in Colletotrichum Gloeosporioides by Solid State Fermentation

Abstract Views: 564  |  PDF Views: 527

Authors

V. Balaji
Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai-600 025, Tamil Nadu, India
P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai-600 025, Tamil Nadu, India

Abstract


Twenty two endophytic fungi isolated from oil seeds were screened for their ability to produce extracellular lipolytic enzymes in solid state fermentation. Among them, the most productive strain, identified as Colletotrichum gloeosporioides was cultivated for lipase production in solid state fermentation (SSF) using oil-mill residue as a solid support. The elaboration of extracellular lipases from C. gloeosporioides on different residual cheap oil substrates was tested. Among these, pongamia oil cake (POC) showed maximum lipase activity (983 U/g dry matter (DM)) followed by coconut oil cake (COC) (925 U/g DM). These two oil industrial residues were chosen for enhanced lipase production. The maximum enzyme yield was obtained when POC was impregnated with CDB (Czapek-Dox Broth) in the ratio of 1:1.5 and with (906 U/g DM). Triton X 100 as the extractant from the fermented solids yielded 1020 U/g DM. The optimum temperature for lipase secretion by C. gloeosporioides was found to be 25°C. Xylose as the carbon source and peptone as the nitrogen source were favorable for the secretion of the enzymes. Tween 60 served as a good lipid substrate for lipase production (1170U/g DM) when POC was the substrate. Amending the POC with Magnesium sulphate as a metallic ion source induced good lipase activity (1240 U/g DM) by C. gloeosporioides. The Sunflower oil was found to be best co-vegetable oil substrate to induce the enzyme (2560 U/g DM). Triton X100 served as the surfactant for lipase secretion in C. gloeosporioides. Thus, pongamia oil cake can be used as cheap substrate for enhanced extracellular lipase production by C. gloeosporioides.

Keywords


Lipases, Colletotrichum gloeosporioides, Solid State Fermentation, Pongamia Oil Cake, Coconut Oil Cake

References





DOI: https://doi.org/10.17485/ijst%2F2008%2Fv1i7%2F29600