Characterizing Key Factors That Influence Recalcitrance of Lignocellulosic Biomass Deconstruction via in-situ liquid IR SNOM Approach
EMSL Project ID
60163
Abstract
Call Topic: This proposal is in response to Functional and Systems Biology on EMSL’s suite of surface science capabilities are ideal for chemical imaging organic matter mineral interactions. Capabilities include in-situ liquid IR SNOM. Specific Aims The most expensive operations in biological processing of cellulosic biomass to fuels and chemicals are for releasing sugars from this naturally recalcitrant material. Unfortunately, such bioprocessing technologies have not yet been commercialized, at least partly because high enzyme doses are currently required to hydrolyze cellulose to glucose at the high yields vital to economic success and to compensate for the rapid fall-off in hydrolysis rate as conversion progresses [1]. Infrared scattering-scanning near-field optical microscopy (IR s-SNOM) can provide chemical information with nanoscale spatial resolution and high sensitivity by combining infrared spectroscopy with atomic force microscopy (AFM). Briefly, the apex of an AFM tip is illuminated with infrared radiation and probes the infrared absorption of a sample with nanoscale spatial resolution. This allows infrared spectroscopy with far higher spatial resolution than conventional techniques and can be used to create chemical images with ~10 nm spatial resolution.[2] This technology will be applied to reacted cellulose to uncover spatial variations of the chemical changes during enzymatic hydrolysis of cellulose correlated with surface morphology. The new capability extends IR s-SNOM to liquid environments, which will allow in-situ measurements of cellulose hydrolysis. Using a novel, bottom illumination geometry, in-liquid IR s-SNOM will probe chemical changes throughout the cellulose hydrolysis process in-situ.[3] Thus, this project seeks to use ultrasensitive IR s-SNOM imaging to quantitatively study enzymatic deconstruction of cellulosic biomass across multiple spatial and temporal scales in order to discover groundbreaking new mechanisms of the enzymatic hydrolysis process. Mission Relevance: The mechanisms of hydrolysis reactions and factors that limit hydrolysis effectiveness remain unclear, and consequently it limits many promising applications in the real world. In particular, the mechanism of rapid decline of cellulose hydrolysis rate during enzymatic hydrolysis, which contributes to high enzyme demands and high cost of biomass bioconversion, is not understood despite of decades of research efforts. Work Plan Overall, this research is to apply innovative techniques to develop a fundamental understanding of the relationship between dynamic change of substrate structure and the functionality of various cellulases components during enzymatic hydrolysis of cellulose. Effects of enzyme-substrate interactions on reaction rates will be studied using key cellulase components from wild type Trichoderma reesei and its variants with different levels of N-linked glycosylation (heterologously expressed in Aspergillus niger) with cellulose substrates provided by WSU.
Project Details
Project type
Exploratory Research
Start Date
2021-12-01
End Date
2023-06-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members