脂肪酶是继蛋白酶和淀粉酶之后的第三大工业化酶制剂,可作为油脂改性的重要催化剂。寻找适合在中性低温环境下应用于油脂深加工的新型脂肪酶具有重要工业应用意义。该研究从黑曲霉基因组中克隆并异源表达了一种分子质量为59 kDa的脂肪酶AtglF,在摇瓶发酵条件下,酶活达到25.36 U/mL。此酶在40 ℃和pH 6.0下表现出最高酶活力,在25~50 ℃和pH 5.0~7.0能够表现出60%以上酶活力;常见金属离子对该酶无显著激活作用,Fe3+和Al3+对该酶具有明显抑制作用;N,N-二甲基甲酰胺、甲苯或甲醇可显著提高酶活力。该酶对所测试的18种天然油脂和人工合成底物皆具有不同程度的水解作用,其中对棕榈油的水解作用最强,其最佳化学合成底物为对硝基苯酚辛酸酯(C8)。以上特性表明,AtglF在食品工业、洗涤剂、有机合成等方面可能具有潜在应用价值。
Lipase is the third largest industrial biocatalyst, following protease and amylase.It has been widely used for oil modification in different fields.Exploring new lipases has garnered significant industrial attention, particularly in industries requiring neutral and low-temperature conditions.In this study, a novel lipase gene atglF (encoding a 59 kDa protein) from Aspergillus niger was cloned and heterologously expressed in Pichia pastoris.Under shaking flask conditions, the maximum activity reached 25.36 U/mL.Recombinant AtglF exhibited optimum activity at 40 ℃ and pH 6.0, retaining over 60% of its maximal activity at 25-50 ℃ and pH 5.0-7.0.No significant activation effect on AtglF activity was observed among the tested metal ions.However, Fe3+ and Al3+ exhibited inhibitory effects on AtglF.The activities were significantly enhanced in the presence of N,N-dimethylformamide, toluene, and methanol.The AtglF displayed different hydrolysis activities toward 18 tested natural oils and synthetic substrates.The highest activities were observed with palm oil and 4-nitrophenyl octanoate (C8), respectively.These characteristics of AtglF indicated its great potential for different industrial applications such as the food industry, detergent industry, and organic synthesis.
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