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Lipase Production by Botryosphaeria ribis EC-01 on Soybean and Castorbean Meals: Optimization, Immobilization, and Application for Biodiesel Production
Authors:Milena M Andrade  Aneli M Barbosa  Matheus R Bofinger  Robert F H Dekker  Josana M Messias  Carmen L B Guedes  Tiago Zaminelli  Bruno H de Oliveira  Valéria M G de Lima  Luiz H Dall’Antonia
Institution:1. Departamento de Química, CCE, Universidade Estadual de Londrina, 86051-990, Londrina, Parana, Brazil
2. Departamento de Bioquímica e Biotecnologia, CCE-Universidade Estadual de Londrina, 86051-990, Londrina, Parana, Brazil
3. Biorefining Research Institute, Lakehead University, Thunder Bay, ON, P7B 5E1, Canada
5. Departamento de Bioquímica e Imunologia, FMRP-Universidade de S?o Paulo, 14049-900, Ribeir?o Preto, S?o Paulo, Brazil
4. Departamento de Ciências Biológicas, Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP), 19806-900, Assis, S?o Paulo, Brazil
Abstract:The effects of soybean and castorbean meals were evaluated separately, and in combinations at different ratios, as substrates for lipase production by Botryosphaeria ribis EC-01 in submerged fermentation using only distilled water. The addition of glycerol analytical grade (AG) and glycerol crude (CG) to soybean and castorbean meals separately and in combination, were also examined for lipase production. Glycerol-AG increased enzyme production, whereas glycerol-CG decreased it. A 24 factorial design was developed to determine the best concentrations of soybean meal, castorbean meal, glycerol-AG, and KH2PO4 to optimize lipase production by B. ribis EC-01. Soybean meal and glycerol-AG had a significant effect on lipase production, whereas castorbean meal did not. A second treatment (22 factorial design central composite) was developed, and optimal lipase production (4,820 U/g of dry solids content (ds)) was obtained when B. ribis EC-01 was grown on 0.5 % (w/v) soybean meal and 5.2 % (v/v) glycerol in distilled water, which was in agreement with the predicted value (4,892 U/g ds) calculated by the model. The unitary cost of lipase production determined under the optimized conditions developed ranged from US$0.42 to 0.44 based on nutrient costs. The fungal lipase was immobilized onto Celite and showed high thermal stability and was used for transesterification of soybean oil in methanol (1:3) resulting in 36 % of fatty acyl alkyl ester content. The apparent K m and V max were determined and were 1.86 mM and 14.29 μmol min?1 mg?1, respectively.
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