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通过易于使用的傅里叶变换红外光谱法监测酶促酯交换反应。

Enzymatic transesterification monitored by an easy-to-use Fourier transform infrared spectroscopy method.

机构信息

Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy.

出版信息

Biotechnol J. 2013 Jan;8(1):133-8. doi: 10.1002/biot.201200173. Epub 2012 Aug 15.

Abstract

Transesterification of triglycerides with short chain alcohols is the key reaction in biodiesel production, in addition to other applications in chemical synthesis. However, it is crucial to optimize reaction conditions to make enzymatic transesterification a cost-effective and competitive process. In this work, a new, easy Fourier transform infrared (FTIR) spectroscopic approach for monitoring the transesterification reaction is reported and compared with a gas-chromatographic method. The concentration of the total methyl esters in the reaction mixture is determined from the peak intensity at ∼1435 cm(-1) in the second derivatives of the FTIR absorption spectra using a linear regression calibration. Interestingly, we found that the use of second derivatives allows an accurate determination of the methyl esters without the interference of free fatty acids. Moreover, information on substrate hydrolysis can be obtained within the same measurement by the infrared absorption at ∼1709 cm(-1) . We applied this approach to monitor methanolysis and hydrolysis reactions catalyzed by different commercial lipases, which displayed different sensitivities to methanol inhibition. Therefore, the FTIR approach reported in this work represents a rapid, inexpensive, and accurate method to monitor enzymatic transesterification, requiring very limited sample preparation and a simple statistical analysis of the spectroscopic data.

摘要

短链醇与甘油三酯的酯交换反应是生物柴油生产的关键反应,此外在化学合成中还有其他应用。然而,优化反应条件对于使酶促酯交换成为一种具有成本效益和竞争力的工艺至关重要。在这项工作中,报道了一种新的、简单的傅里叶变换红外(FTIR)光谱方法来监测酯交换反应,并与气相色谱法进行了比较。通过在 FTIR 吸收光谱的二阶导数中使用线性回归校准,从约 1435 cm(-1) 处的峰强度确定反应混合物中总甲酯的浓度。有趣的是,我们发现二阶导数的使用允许在没有游离脂肪酸干扰的情况下准确测定甲酯。此外,通过在约 1709 cm(-1) 处的红外吸收,可以在同一测量中获得关于底物水解的信息。我们将这种方法应用于监测不同商业脂肪酶催化的甲醇解和水解反应,这些脂肪酶对甲醇抑制的敏感性不同。因此,本文报道的 FTIR 方法代表了一种快速、廉价和准确的监测酶促酯交换的方法,仅需要非常有限的样品制备和对光谱数据的简单统计分析。

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