Departamento de Química, CCE, Universidade Estadual de Londrina, 86051-990 Londrina, Parana, Brazil.
Appl Biochem Biotechnol. 2013 Aug;170(7):1792-806. doi: 10.1007/s12010-013-0309-9. Epub 2013 Jun 9.
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 2(4) 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 (2(2) 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.
分别评估了大豆和蓖麻粕作为唯一用水进行的液体发酵中脂肪酶生产的底物,以及以不同比例组合使用时的效果。还分别和组合添加分析级甘油(AG)和粗甘油(CG),考察它们对脂肪酶生产的影响。AG 甘油增加了酶的产量,而 CG 甘油则降低了产量。采用 2(4)析因设计确定大豆粕、蓖麻粕、AG 甘油和 KH2PO4 的最佳浓度,以优化 B. ribis EC-01 的脂肪酶生产。大豆粕和 AG 甘油对脂肪酶生产有显著影响,而蓖麻粕则没有。设计了第二个处理(2(2)析因设计中心组合),当 B. ribis EC-01 在 0.5%(w/v)大豆粕和 5.2%(v/v)蒸馏水中生长时,获得了最佳的脂肪酶生产(4,820 U/g 干物质含量(ds)),与模型计算的预测值(4,892 U/g ds)一致。在优化条件下确定的脂肪酶生产成本,基于营养成本,从每单位 0.42 美元到 0.44 美元不等。固定化在 Celite 上的真菌脂肪酶具有较高的热稳定性,并用于在甲醇(1:3)中进行大豆油的转酯化,生成 36%的脂肪酸烷酯含量。测定了表观 K m 和 V max,分别为 1.86 mM 和 14.29 μmol min(-1) mg(-1)。