Engineered catalytic biofilms: Site-specific enzyme immobilization onto E-coli curli nanofibers

被引:105
作者
Botyanszki, Zsofia [1 ,2 ]
Tay, Pei Kun R. [2 ,3 ]
Nguyen, Peter Q. [2 ,3 ]
Nussbaumer, Martin G. [2 ,3 ]
Joshi, Neel S. [2 ,3 ]
机构
[1] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA
[2] Harvard Univ, Wyss Inst Biol Inspired Engn, Cambridge, MA 02138 USA
[3] Harvard Univ, Sch Engn & Appl Sci, Cambridge, MA 02138 USA
关键词
biofilm; bacterial immobilization; curli fibers; biocatalysis; enzyme display; extracellular matrix; MICROBIAL BIOFILMS; BACTERIAL DISPLAY; ORGANIC MEDIA; BIOCATALYSIS; CELLS; PROTEIN; CHEMICALS; ALGINATE; SYSTEM; MATRIX;
D O I
10.1002/bit.25638
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Biocatalytic transformations generally rely on purified enzymes or whole cells to perform complex transformations that are used on industrial scale for chemical, drug, and biofuel synthesis, pesticide decontamination, and water purification. However, both of these systems have inherent disadvantages related to the costs associated with enzyme purification, the long-term stability of immobilized enzymes, catalyst recovery, and compatibility with harsh reaction conditions. We developed a novel strategy for producing rationally designed biocatalytic surfaces based on Biofilm Integrated Nanofiber Display (BIND), which exploits the curli system of E. coli to create a functional nanofiber network capable of covalent immobilization of enzymes. This approach is attractive because it is scalable, represents a modular strategy for site-specific enzyme immobilization, and has the potential to stabilize enzymes under denaturing environmental conditions. We site-specifically immobilized a recombinant -amylase, fused to the SpyCatcher attachment domain, onto E. coli curli fibers displaying complementary SpyTag capture domains. We characterized the effectiveness of this immobilization technique on the biofilms and tested the stability of immobilized -amylase in unfavorable conditions. This enzyme-modified biofilm maintained its activity when exposed to a wide range of pH and organic solvent conditions. In contrast to other biofilm-based catalysts, which rely on high cellular metabolism, the modified curli-based biofilm remained active even after cell death due to organic solvent exposure. This work lays the foundation for a new and versatile method of using the extracellular polymeric matrix of E. coli for creating novel biocatalytic surfaces. Biotechnol. Bioeng. 2015;112: 2016-2024. (c) 2015 Wiley Periodicals, Inc.
引用
收藏
页码:2016 / 2024
页数:9
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