College of Food Science and Nutrition Engineering, China Agriculture University, Beijing 100083, China.
College of Engineering, China Agriculture University, Beijing 100083, China.
Food Chem. 2021 Jul 15;350:129212. doi: 10.1016/j.foodchem.2021.129212. Epub 2021 Feb 2.
A novel alkaline cold-active phospholipase C (PLC) gene (AoPC) from Aspergillus oryzae was cloned. AoPC exhibited the highest sequence similarity of 32.5% with that of a PLC from Arabidopsis thaliana. The gene was co-expressed in Pichia pastoris with molecular chaperone PDI (protein disulfide isomerases), and the highest PLC activity of 82, 782 U mL was achieved in a 5-L fermentor. The recombinant enzyme (AoPC) was most active at pH 8.0 and 25 °C, respectively, and it was stable over a broad pH range of 4.5-9.0 and up to 40 °C. It is the first fungal alkaline PLC. The application of AoPC (with 25% citric acid, w/w) in oil degumming process significantly reduced the phosphorus of crude soybean oil by 93.3% to a commercially acceptable level (<10 mg kg). Therefore, the relatively high yield and excellent properties of AoPC may possess it great potential in crude oil refining industry.
从米曲霉中克隆到一种新型碱性冷活性磷脂酶 C（PLC）基因（AoPC）。AoPC 与拟南芥 PLC 的序列相似性最高，为 32.5%。该基因与分子伴侣 PDI（蛋白质二硫键异构酶）在毕赤酵母中共同表达，在 5-L 发酵罐中获得了最高的 82,782 U/mL PLC 活性。重组酶（AoPC）在 pH8.0 和 25°C 时活性最高，在 pH4.5-9.0 范围内稳定，最高可达 40°C。这是第一个真菌碱性 PLC。AoPC（添加 25%柠檬酸，w/w）在油脂脱胶过程中的应用使粗大豆油中的磷含量降低了 93.3%，达到了商业上可接受的水平（<10mg/kg）。因此，AoPC 较高的产率和优良的性质可能使其在原油精炼行业具有巨大的潜力。
