EAGER: Rapid Development of the "Rosettazyme" - A Novel Enzyme Sequestration Platform for Cellulosic Substrate Deconstruction

R. Michael CeballosProposal#0929484EAGER：快速开发“Rosettazyme”-一种用于纤维素底物分解的新型酶螯合平台目前生物乙醇生产技术受到有效分解纤维素生物质为可发酵糖的能力的限制。 此外，纤维素底物的许多预处理方案需要酸性洗涤和高温工艺，这抑制了生物乙醇生产工艺中使用的纤维素降解酶的催化效率。 利用来自火山温泉的超嗜热嗜酸微生物的蛋白质，该项目的研究人员开发了一种耐温耐酸的酶螯合平台，能够结合多种纤维素分解酶形成一种称为“rosettazyme”的“超级酶”。 在初步研究中，已经表明这些rosettazyme可以将纤维素底物的简单糖生产提高两倍以上。 该项目的目的是使用来自几个物种的结合的木质纤维素解构酶的各种组合来进一步开发这种新技术，以使纤维素分解/转化效率提高5至10倍。 底物解构效率的这种水平的改进已经被确定为使该技术成为当前生物乙醇生产方法的经济上可行的替代方案的阈值。 这项拟议中的工作将是美国几个学术机构和政府实验室的共同努力。 该项目还将为来自代表性不足的群体和经济上服务不足的背景的理科学生提供蛋白质生物化学，微生物学，遗传工程和替代能源科学的研究经验。在该项目的两年内，研究人员将使用细菌，古细菌和/或真菌酶开发几种rosettazyme亚型，以提高木质纤维素转化为简单可发酵糖的效率。

R. Michael CeballosProposal#0929484EAGER: Rapid Development of the 'Rosettazyme' - A Novel Enzyme Sequestration Platform for Cellulosic Substrate DeconstructionCurrent technology in bioethanol production is limited by the ability to efficiently breakdown cellulosic biomass into fermentable sugars. In addition, many pre-treatment regimens for cellulosic substrate require acidic washes and high temperature processes that inhibit the catalytic efficiency of cellulose-degrading enzymes used in bioethanol production processes. Using proteins derived from hyperthermoacidophilic microbes from volcanic hot springs, the investigators on this project have developed a temperature- and acid-resistant enzyme sequestration platform capable of binding multiple cellulose-deconstruction enzymes to form a 'super-enzyme', called a 'rosettazyme'. In preliminary studies, it has been shown that these rosettazymes can improve simple sugar production from cellulosic substrates by more than two-fold. The purpose of this project is to further develop this novel technology using various combinations of bound lignocellulosic deconstruction enzymes from several species in order to generate a 5 to 10-fold increase in cellulose breakdown/conversion efficiency. This level of improvement in substrate deconstruction efficiency has been determined to be the threshold for making this technology an economically viable alternative to current bioethanol production methods. The proposed work will be a joint effort between several academic institutions and government laboratories throughout the United States. This project will also serve to provide science students from underrepresented groups and economically underserved backgrounds with research experiences in protein biochemistry, microbiology, genetic engineering, and alternative energy sciences. Within the two years of this project, the investigators will develop several rosettazyme subtypes using bacterial, archaeal, and/or fungal enzymes to increase the efficiency of lignocelluose conversion to simple fermentable sugars.