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

R. Michael ceballos提案#0929484EAGER：“Rosettazyme”的快速发展-一种用于纤维素底物解构的新型酶隔离平台目前生物乙醇生产的技术受到有效将纤维素生物质分解为可发酵糖的能力的限制。此外，许多纤维素底物的预处理方案需要酸性洗涤和高温处理，这抑制了生物乙醇生产过程中使用的纤维素降解酶的催化效率。利用来自火山温泉的高温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隔离平台，能够结合多种纤维素分解酶形成一种“超级酶”，称为“罗塞塔酶”。在初步研究中，已经证明这些玫瑰酶可以将纤维素底物的单糖产量提高两倍以上。该项目的目的是进一步开发这种新技术，使用来自几种物种的结合木质纤维素分解酶的各种组合，以使纤维素分解/转化效率提高5到10倍。这种底物分解效率的提高水平已被确定为使该技术成为目前生物乙醇生产方法经济上可行的替代方法的门槛。这项提议的工作将由美国各地的几个学术机构和政府实验室共同努力。该项目还将为来自代表性不足群体和经济条件不足背景的理科生提供蛋白质生物化学、微生物学、基因工程和替代能源科学方面的研究经验。在这个项目的两年内，研究人员将利用细菌、古细菌和/或真菌酶开发几种罗塞塔酶亚型，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.