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Real-Time Measurement of Cellobiose and Glucose Formation during Enzymatic Biomass Hydrolysis
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-05-20 , DOI: 10.1021/acs.analchem.1c01182 Hucheng Chang 1 , Lena Wohlschlager 1 , Florian Csarman 1 , Adrian Ruff 2 , Wolfgang Schuhmann 2 , Stefan Scheiblbrandner 1 , Roland Ludwig 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-05-20 , DOI: 10.1021/acs.analchem.1c01182 Hucheng Chang 1 , Lena Wohlschlager 1 , Florian Csarman 1 , Adrian Ruff 2 , Wolfgang Schuhmann 2 , Stefan Scheiblbrandner 1 , Roland Ludwig 1
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Enzymatic hydrolysis of lignocellulosic biomass for biofuel production relies on complex multi-enzyme ensembles. Continuous and accurate measurement of the released key products is crucial in optimizing the industrial degradation process and also investigating the activity and interaction between the involved enzymes and the insoluble substrate. Amperometric biosensors have been applied to perform continuous cellobiose measurements during the enzymatic hydrolysis of pure cellulose powders. The oxygen-sensitive mediators used in these biosensors restricted their function under physiological or industrial conditions. Also, the combined measurements of the hydrolysis products cellobiose and glucose require a high selectivity of the biorecognition elements. We employed an [Os(2,2′-bipyridine)2Cl]Cl-modified polymer and cellobiose dehydrogenase to fabricate a cellobiose biosensor, which can accurately and specifically detect cellobiose even in the presence of oxygen and the other main product glucose. Additionally, a glucose biosensor was fabricated to simultaneously measure glucose produced from cellobiose by β-glucosidases. The cellobiose and glucose biosensors work at applied potentials of +0.25 and +0.45 V versus Ag|AgCl (3 M KCl), respectively, and can selectively detect their substrate. Both biosensors were used in combination to monitor the hydrolysis of pure cellulose of low crystallinity or industrial corncob samples. The obtained results correlate with the high-performance liquid chromatography pulsed amperometric detection analysis and demonstrate that neither oxygen nor the presence of redox-active compounds from the lignin fraction of the corncob interferes with the measurements.
中文翻译：
酶促生物质水解过程中纤维二糖和葡萄糖形成的实时测量
用于生物燃料生产的木质纤维素生物质的酶促水解依赖于复杂的多酶集合。连续准确地测量释放的关键产物对于优化工业降解过程以及研究相关酶与不溶性底物之间的活性和相互作用至关重要。电流生物传感器已被应用于在纯纤维素粉末的酶促水解过程中进行连续纤维二糖测量。这些生物传感器中使用的氧敏感介质限制了它们在生理或工业条件下的功能。此外，水解产物纤维二糖和葡萄糖的组合测量需要生物识别元件的高选择性。我们采用 [Os(2,2'-bipyridine) 2Cl]Cl 修饰的聚合物和纤维二糖脱氢酶制备纤维二糖生物传感器，即使在氧气和其他主要产物葡萄糖存在的情况下也能准确、特异性地检测纤维二糖。此外，还制造了一个葡萄糖生物传感器，以同时测量 β-葡萄糖苷酶从纤维二糖产生的葡萄糖。纤维二糖和葡萄糖生物传感器分别在 +0.25 和 +0.45 V 与 Ag|AgCl (3 M KCl) 的外加电位下工作，并且可以选择性地检测它们的底物。两种生物传感器结合使用来监测低结晶度或工业玉米芯样品的纯纤维素的水解。
更新日期：2021-06-01
中文翻译：
酶促生物质水解过程中纤维二糖和葡萄糖形成的实时测量
用于生物燃料生产的木质纤维素生物质的酶促水解依赖于复杂的多酶集合。连续准确地测量释放的关键产物对于优化工业降解过程以及研究相关酶与不溶性底物之间的活性和相互作用至关重要。电流生物传感器已被应用于在纯纤维素粉末的酶促水解过程中进行连续纤维二糖测量。这些生物传感器中使用的氧敏感介质限制了它们在生理或工业条件下的功能。此外，水解产物纤维二糖和葡萄糖的组合测量需要生物识别元件的高选择性。我们采用 [Os(2,2'-bipyridine) 2Cl]Cl 修饰的聚合物和纤维二糖脱氢酶制备纤维二糖生物传感器，即使在氧气和其他主要产物葡萄糖存在的情况下也能准确、特异性地检测纤维二糖。此外，还制造了一个葡萄糖生物传感器，以同时测量 β-葡萄糖苷酶从纤维二糖产生的葡萄糖。纤维二糖和葡萄糖生物传感器分别在 +0.25 和 +0.45 V 与 Ag|AgCl (3 M KCl) 的外加电位下工作，并且可以选择性地检测它们的底物。两种生物传感器结合使用来监测低结晶度或工业玉米芯样品的纯纤维素的水解。
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