Badgujar Kirtikumar C, Bhanage Bhalchandra M
Department of Chemistry, Institute of Chemical Technology , Mumbai-400 019, India.
J Phys Chem B. 2014 Dec 26;118(51):14808-19. doi: 10.1021/jp5093493. Epub 2014 Dec 16.
In the present study, we have synthesized a biocompatible hybrid carrier of hypromellose (HY) and poly(vinyl alcohol) (PVA) for immobilization of Burkholderia cepacia lipase (BCL). The immobilized biocatalyst HY:PVA:BCL was subjected to determination of half-life time (τ) and deactivation rate constant (K(D)) in various organic solvents. Biocatalyst showed higher τ-value in a nonpolar solvent like cyclohexane (822 h) as compared to that of a polar solvent such as acetone (347 h), which signifies better compatibility of biocatalyst in the nonpolar solvents. Furthermore, the K(D)-value was found to be less in cyclohexane (0.843 × 10(-3)) as compared to acetone (1.997 × 10(-3)), indicating better stability in the nonpolar solvents. Immobilized-BCL (35 mg) was sufficient to achieve 99% conversion of phenethyl butyrate (natural constituent of essential oils and has wide industrial applications) using phenethyl alcohol (2 mmol) and vinyl butyrate (6 mmol) at 44 °C in 3 h. The activation energy (E(a)) was found to be lower for immobilized-BCL than crude-BCL, indicating better catalytic efficiency of immobilized lipase BCL. The immobilized-BCL reported 6-fold superior biocatalytic activity and 8 times recyclability as compared to crude-BCL. Improved catalytic activity of immobilized enzyme in nonpolar media was also supported by thermodynamic activation parameters such as enthalpy (ΔH(⧧)), entropy (ΔS(⧧)) and Gibb's free energy (ΔG(⧧)) study, which showed that phenethyl butyrate synthesis catalyzed by immobilized-BCL was feasible as compared to crude-BCL. The present work explains a thermodynamic investigation and superior biocatalytic activity for phenethyl butyrate synthesis using biocompatible immobilized HY:PVA:BCL in nonaqueous media for the first time.
在本研究中,我们合成了一种由羟丙甲纤维素(HY)和聚乙烯醇(PVA)组成的生物相容性杂化载体,用于固定洋葱伯克霍尔德菌脂肪酶(BCL)。将固定化生物催化剂HY:PVA:BCL置于各种有机溶剂中测定半衰期(τ)和失活速率常数(K(D))。与极性溶剂如丙酮(347小时)相比,生物催化剂在非极性溶剂如环己烷(822小时)中显示出更高的τ值,这表明生物催化剂在非极性溶剂中具有更好的相容性。此外,发现环己烷中的K(D)值(0.843×10(-3))低于丙酮中的K(D)值(1.997×10(-3)),表明在非极性溶剂中具有更好的稳定性。固定化BCL(35毫克)足以在44℃下3小时内使用苯乙醇(2毫摩尔)和丁酸乙烯酯(6毫摩尔)实现99%的丁酸苯乙酯(精油的天然成分,具有广泛的工业应用)转化。发现固定化BCL的活化能(E(a))低于粗BCL,表明固定化脂肪酶BCL具有更好的催化效率。与粗BCL相比,固定化BCL的生物催化活性高6倍,可重复使用8次。固定化酶在非极性介质中催化活性的提高也得到了热力学活化参数如焓(ΔH(⧧))、熵(ΔS(⧧))和吉布斯自由能(ΔG(⧧))研究的支持,这些研究表明与粗BCL相比,固定化BCL催化丁酸苯乙酯合成是可行的。本工作首次解释了使用生物相容性固定化HY:PVA:BCL在非水介质中合成丁酸苯乙酯的热力学研究和优异的生物催化活性。