Departament of Biochemistry and Chemical Technology, IQ/UNESP, Rua Prof. Francisco Degni, 55, CEP, Araraquara, SP, 14800-060, Brazil.
Department of Chemistry and Environmental Sciences, IBILCE/UNESP, Rua Cristóvão Colombo, 2265, CEP, São José do Rio Preto, SP, 15054-000, Brazil.
Appl Biochem Biotechnol. 2021 Jan;193(1):33-51. doi: 10.1007/s12010-020-03411-w. Epub 2020 Aug 18.
Fungal enzymes are widely used in technological processes and have some interesting features to be applied in a variety of biosynthetic courses. Here, free and substrate-immobilised lipases from Fusarium verticillioides P24 were obtained by solid-state fermentation using wheat bran as substrate and fungal carrier. Based on their hydrolytic and transesterification activities, the lipases were characterised as pH-dependent in both reactions, with higher substrate conversion in an alkaline environment. Thermally, the lipases performed well from 30 to 45 °C, being more stable in mild conditions. Organic solvents significantly influenced the lipase selectivity using different vegetable oils as fatty acid source. Omega(ω)-3 production in n-hexane achieved 45% using canola oil, against ≈ 18% in cyclohexane. However, ω-6 production was preferably produced for both solvents using linseed oil with significant alterations in the yield (≈ 79% and 49% for n-hexane and cyclohexane, respectively). Moreover, the greatest enzyme selectivity for ω-6 led us to suppose a lipase preference for the Sn1 position of the triacylglycerol. Lastly, a transesterification reaction was performed, achieving 90% of ester conversion in 72 h. This study reports the characterisation and use of free and substrate-immobilised lipases from Fusarium verticillioides P24 as an economic and efficient method for the first time.
真菌酶广泛应用于技术过程中，具有一些有趣的特点，可应用于各种生物合成过程。本文采用固态发酵法，以麦麸为底物和真菌载体，获得了Verticillium 禾谷镰孢 P24 的游离和固定化脂肪酶。基于其水解和转酯化活性，这些脂肪酶在两种反应中均表现出 pH 依赖性，在碱性环境中具有更高的底物转化率。从 30 到 45°C，脂肪酶的热稳定性良好，在温和条件下更稳定。有机溶剂对不同植物油作为脂肪酸源时脂肪酶的选择性有显著影响。使用菜籽油在正己烷中ω-3 的产量达到 45%，而在环己烷中约为 18%。然而，对于这两种溶剂，亚麻籽油都更有利于ω-6 的生成，产率有显著改变（正己烷和环己烷分别约为 79%和 49%）。此外，脂肪酶对ω-6 的最大选择性使我们假设它对三酰基甘油的 Sn1 位置具有偏好。最后，进行了转酯化反应，72 小时内达到 90%的酯转化率。本研究首次报道了 Verticillium 禾谷镰孢 P24 的游离和固定化脂肪酶的特性和用途，这是一种经济高效的方法。
