Laboratory of Biochemistry and Enzymatic Engineering of Lipases, National School of Engineers of Sfax, University of Sfax, Sfax, Tunisia.
Department of Chemical Engineering, College of Engineering, Imam Mohammad Ibn Saud Islamic University, IMSIU, Riyadh, Saudi Arabia.
Prep Biochem Biotechnol. 2023 Nov;53(10):1165-1175. doi: 10.1080/10826068.2023.2177867. Epub 2023 Feb 15.
This work deals with the optimization of an extracellular phospholipase C production by (PLC) using Response Surface Methodology (RMS) and Box-Behnken design. In fact, after optimization, a maximum phospholipase activity (51 U/ml) was obtained after 6 h of cultivation on tryptone (10 g/L), yeast extract (10 g/L), NaCl (8.125 g/L), pH 7.5 with initial OD (0.15). The PLC activity, esteemed by the model (51 U) was very approximate to activity gutted experimentally (50 U). The PLC can be considered as thermoactive phospholipase since it showed a maximal activity of 50 U/mL at 60 °C using egg yolk or egg phosphatidylcholine (PC) as substrate. In addition, the enzyme was active at pH 7 and is stable after incubation at 55 °C for 30 min. The application of phospholipase C in soybean oil degumming was investigated. Our results showed that when using enzymatic degumming, the residual phosphorus decrease more than with water degumming, indeed, it passes from 718 ppm in soybean crude oil to 100 ppm and 52 ppm by degumming using water and enzymatic process, respectively. The diacylgycerol (DAG) yield showed an increase of 1.2% with enzymatic degumming compared to soybean crude oil. This makes our enzyme a potential candidate for food industrial applications such as enzymatic degumming of vegetable oils.
本文采用响应面法(RMS)和 Box-Behnken 设计优化了(PLC)细胞外磷脂酶 C 的生产。事实上,经过优化后,在含有 10g/L 胰蛋白胨、10g/L 酵母提取物、8.125g/L NaCl、初始 OD(0.15)、pH7.5 的条件下培养 6 小时后,获得了最大的磷脂酶活性(51U/ml)。通过模型估计的 PLC 活性(51U)非常接近实验测定的活性(50U)。该 PLC 可视为热活性磷脂酶,因为它在 60°C 时使用蛋黄或卵磷酯(PC)作为底物时表现出最大活性 50U/mL。此外,该酶在 pH7 下具有活性,在 55°C 孵育 30 分钟后保持稳定。本文还研究了磷脂酶 C 在大豆油脱胶中的应用。结果表明,与水脱胶相比,酶法脱胶时残磷含量降低更多,用酶法和水法脱胶后,粗大豆油中的磷含量分别从 718ppm 降至 100ppm 和 52ppm。与粗大豆油相比,二酰基甘油(DAG)的产量通过酶法脱胶增加了 1.2%。这使得我们的酶成为食品工业应用的潜在候选物,如植物油的酶法脱胶。