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Comparison of analytical methods for rapid & reliable quantification of plant-based carbohydrates for the quintessential bioenergy educator
bioRxiv - Scientific Communication and Education Pub Date : 2020-05-23 , DOI: 10.1101/2020.05.21.106468
Shishir P. S. Chundawat

Biochemical conversion of plant-based insoluble carbohydrate polymers, such as starch from corn grains or cellulose-hemicellulose from corn stover, into soluble fermentable sugars (e.g., glucose and xylose) for bioenergy production has seen tremendous research activity and commercial-scale biorefineries deployment over the last three decades, particularly in regions around the world that have a dominant agricultural-based economy. Therefore, educators in schools and universities have developed various hands-on experimental activities to engage the general public and students either in outreach events or lab/classroom-based settings to instruct students on various inter-disciplinary concepts relevant to bioenergy and biochemicals production. One of the limitations of most available protocols is the lack of systematic and comprehensive comparison of educator-friendly analytical tools and protocols for quantitative analysis of water-soluble carbohydrates commonly encountered in a biorefinery backdrop during the biochemical conversion of lignocellulosic biomass to biofuels/biochemicals. Here, we systematically compare and validate findings from four leading analytical approaches for detection and quantification of lignocellulosic biomass derived soluble carbohydrates. We compare these assay methods based on the overall ease of use, detection accuracy/sensitivity, commercial availability, analytical cost per assay run, and suitability for use by instructors in biorefining specific hands-on activity protocols. Next, we provide a detailed instructional protocol that utilizes one of these validated soluble sugar assays as part of a 90 min hands-on bioenergy focused activity (called Grass-to-Gas) conducted at Rutgers University with pre-university high school students. Grass-to-Gas activity involves students running biochemical assays that helps them understand the various facets of cellulosic biomass hydrolysis by commercial cellulase enzymes and monitoring the total glucose product released using our validated sugar assays to finally estimate the fractional conversion of cellulose-to-glucose. Lastly, we further demonstrate how such carbohydrate-based analytical methods can be used by instructors to help university students explore and understand various chemistry, biochemistry, and chemical engineering concepts relevant to other advanced operations involved in lignocellulose biorefining. These activity protocols would greatly aid educators teaching interdisciplinary science and engineering concepts to students in the backdrop of lignocellulose biorefining.

中文翻译:

典型生物能源教育者快速可靠地定量植物性碳水化合物的分析方法的比较

将植物基不溶性碳水化合物聚合物(例如玉米粒中的淀粉或玉米秸秆中的纤维素半纤维素)生物化学转化为可发酵的可发酵糖(例如葡萄糖和木糖)用于生物能源生产,已经看到了巨大的研究活动和商业规模的生物炼制在过去的三十年中,尤其是在世界上以农业为基础的经济占主导地位的地区。因此,学校和大学中的教育工作者开展了各种动手实验活动,以使普通公众和学生参与推广活动或实验室/教室设置,以指导学生有关与生物能源和生化试剂生产相关的各种跨学科概念。大多数可用方案的局限性之一是缺乏系统的,全面的比较,即对木质生物质生物质向生物燃料/生物化学物质进行生物化学转化过程中生物精炼厂通常遇到的水溶性碳水化合物进行定量分析的教育者友好的分析工具和方案。在这里,我们系统地比较和验证了四种检测和定量木质纤维素生物质衍生的可溶性碳水化合物的领先分析方法的发现。我们根据总体易用性,检测准确性/灵敏度,商业可用性,每次分析运行的分析成本以及讲师在生物提炼特定动手操作规程中的适用性来比较这些分析方法。下一个,我们提供了详细的教学方案,该方案利用这些经过验证的可溶性糖测定法之一,作为在罗格斯大学与大学预科高中生进行的90分钟动手生物能源重点活动(称为草气)的一部分。从草到气的活动需要学生进行生化分析,以帮助他们了解商业纤维素酶水解纤维素生物质的各个方面,并使用我们经过验证的糖分析方法监测释放的总葡萄糖产物,以最终估算纤维素转化为葡萄糖的分数转化率。最后,我们进一步演示了教师如何使用这种基于碳水化合物的分析方法来帮助大学生探索和理解各种化学,生物化学,与涉及木质纤维素生物精炼的其他高级操作相关的化学工程概念。这些活动协议将极大地帮助教育工作者在木质纤维素生物精炼的背景下向学生讲授跨学科的科学和工程学概念。
更新日期:2020-05-23
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