Jumbri Khairulazhar, Al-Haniff Rozy Mohd Fahruddin, Ashari Siti Efliza, Mohamad Rosfarizan, Basri Mahiran, Fard Masoumi Hamid Reza
Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia.
Enzyme and Microbial Technology Research Centre, Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia.
PLoS One. 2015 Dec 14;10(12):e0144664. doi: 10.1371/journal.pone.0144664. eCollection 2015.
Kojic acid is widely used to inhibit the browning effect of tyrosinase in cosmetic and food industries. In this work, synthesis of kojic monooleate ester (KMO) was carried out using lipase-catalysed esterification of kojic acid and oleic acid in a solvent-free system. Response Surface Methodology (RSM) based on central composite rotatable design (CCRD) was used to optimise the main important reaction variables, such as enzyme amount, reaction temperature, substrate molar ratio, and reaction time along with immobilised lipase from Candida Antarctica (Novozym 435) as a biocatalyst. The RSM data indicated that the reaction temperature was less significant in comparison to other factors for the production of a KMO ester. By using this statistical analysis, a quadratic model was developed in order to correlate the preparation variable to the response (reaction yield). The optimum conditions for the enzymatic synthesis of KMO were as follows: an enzyme amount of 2.0 wt%, reaction temperature of 83.69°C, substrate molar ratio of 1:2.37 (mmole kojic acid:oleic acid) and a reaction time of 300.0 min. Under these conditions, the actual yield percentage obtained was 42.09%, which is comparably well with the maximum predicted value of 44.46%. Under the optimal conditions, Novozym 435 could be reused for 5 cycles for KMO production percentage yield of at least 40%. The results demonstrated that statistical analysis using RSM can be used efficiently to optimise the production of a KMO ester. Moreover, the optimum conditions obtained can be applied to scale-up the process and minimise the cost.
曲酸在化妆品和食品工业中被广泛用于抑制酪氨酸酶的褐变作用。在本研究中,在无溶剂体系中利用脂肪酶催化曲酸与油酸的酯化反应合成了曲酸单油酸酯(KMO)。采用基于中心复合旋转设计(CCRD)的响应面法(RSM)来优化主要反应变量,如酶用量、反应温度、底物摩尔比和反应时间,同时使用南极假丝酵母固定化脂肪酶(诺维信435)作为生物催化剂。RSM数据表明,与其他因素相比,反应温度对KMO酯的生产影响较小。通过这种统计分析,建立了一个二次模型,以便将制备变量与响应(反应产率)相关联。KMO酶促合成的最佳条件如下:酶用量2.0 wt%,反应温度83.69℃,底物摩尔比1:2.37(毫摩尔曲酸:油酸),反应时间300.0分钟。在这些条件下,实际获得的产率百分比为42.09%,与最大预测值44.46%相当。在最佳条件下,诺维信435可重复使用5个循环,KMO生产百分比产率至少为40%。结果表明,使用RSM进行统计分析可以有效地优化KMO酯的生产。此外,获得的最佳条件可应用于扩大生产规模并降低成本。