High-throughput microplate technique for enzymatic hydrolysis of lignocellulosic biomass.

@article{Chundawat2008HighthroughputMT,
  title={High-throughput microplate technique for enzymatic hydrolysis of lignocellulosic biomass.},
  author={Shishir P. S. Chundawat and Venkatesh Balan and Bruce E. Dale},
  journal={Biotechnology and bioengineering},
  year={2008},
  volume={99 6},
  pages={
          1281-94
        }
}
Several factors will influence the viability of a biochemical platform for manufacturing lignocellulosic based fuels and chemicals, for example, genetically engineering energy crops, reducing pre-treatment severity, and minimizing enzyme loading. Past research on biomass conversion has focused largely on acid based pre-treatment technologies that fractionate lignin and hemicellulose from cellulose. However, for alkaline based (e.g., AFEX) and other lower severity pre-treatments it becomes… 

Figures and Tables from this paper

Shotgun Approach to Increasing Enzymatic Saccharification Yields of Ammonia Fiber Expansion Pretreated Cellulosic Biomass
TLDR
It is shown that directly blending crude cellulolytic and hemicellulolytic enzymes-rich microbial secretomes can maximize specific activity on AFEX treated biomass without having to isolate individual enzymes.
Synthetic enzyme mixtures for biomass deconstruction: Production and optimization of a core set
TLDR
The GENPLAT platform and the optimized “core” set together provide a starting point for the rapid testing and optimization of alternate core enzymes from other microbial and recombinant sources as well as for the testing of “accessory” proteins for development of superior enzyme mixtures for biomass conversion.
Application of a microassay method to study enzymatic hydrolysis of pretreated wheat straw
TLDR
This enzymatic microassay represents a rapid method to test the performance of new selected cellulase enzymes on real pretreated lignocellulosic substrates and will enable evaluation of enzyme components separately, or optimized mixtures.
Micro-assay method for enzymatic saccharification of industrially relevant lignocellulose substrates
The development of micro-assays suitable for automation systems represents a helpful alternative by which many enzymes and factors can be studied simultaneously for the conversion of lignocellulose.
High‐Throughput Pretreatment and Hydrolysis Systems for Screening Biomass Species in Aqueous Pretreatment of Plant Biomass
TLDR
The discovery and use of biomass species with reduced recalcitrance, when combined with optimized pretreatment processes and enzyme mixtures, could potentially improve the commercial viability of fuels and chemicals production from lignocellulosic biomass.
Application of high throughput pretreatment and co‐hydrolysis system to thermochemical pretreatment. Part 1: Dilute acid
TLDR
A single‐step neutralization and buffering method was developed to allow effective enzymatic hydrolysis of the whole pretreated slurry, demonstrating the method to be a useful extension of HTPH systems.
High-Throughput Screening Techniques for Biomass Conversion
High-throughput (HTP) screening of biomass or biomass-degrading enzymes, regardless of the desired outcome, is fraught with obstacles and challenges not typically faced in more traditional
Lignin triggers irreversible cellulase loss during pretreated lignocellulosic biomass saccharification
TLDR
An FPLC (fast protein liquid chromatography)-based methodology is utilized to quantify free Trichoderma reesei cellulases concentration within a complex hydrolyzate mixture during the varying time course of biomass saccharification, aiding in further understanding the complex mechanisms of non-productive binding of cellulases to pretreated lignocellulosic biomass.
A High-Throughput Platform for Screening Milligram Quantities of Plant Biomass for Lignocellulose Digestibility
TLDR
A high-throughput digestibility platform (HTDP) for screening collections of germplasm for improved digestibility is described, which was developed under the auspices of the Department of Energy-Great Lakes Bioenergy Research Center (DOE-GLBRC).
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 55 REFERENCES
A rapid microassay to evaluate enzymatic hydrolysis of lignocellulosic substrates
TLDR
A rapid assay for lignocellulose hydrolysis has been developed and shows considerable time and cost benefits over the standard assay protocol and is applicable to a broad range of lignOcellulosic substrates.
In vitro gas production as a surrogate measure of the fermentability of cellulosic biomass to ethanol
TLDR
Because the in vitro ruminal fermentation assay can be performed without sterilization of the medium and does not require aseptic conditions, this assay may be useful for biomass feedstock agronomic and breeding research.
Standard assays do not predict the efficiency of commercial cellulase preparations towards plant materials.
TLDR
It was concluded that the choice of an enzyme preparation is more dependent on the characteristics of the substrate rather than on standard enzyme-activities measured, because there was a large difference in the performance of the fourteen enzyme samples.
Outlook for cellulase improvement: screening and selection strategies.
Potential and utilization of thermophiles and thermostable enzymes in biorefining
TLDR
existing and potential applications of thermophiles and thermostable enzymes with focus on conversion of carbohydrate containing raw materials are discussed and strategies that enhance thermostablity of enzymes both in vivo and in vitro are assessed.
Substrate pretreatment: the key to effective enzymatic hydrolysis of lignocellulosics?
TLDR
Focusing particularly on steam pretreatment, this review examines the influence that pretreatment conditions have on substrate characteristics such as lignin and hemicellulose content, crystallinity, degree of polymerization and specific surface, and the resulting implications for effective hydrolysis by cellulases.
Optimization of Ammonia Fiber Expansion (AFEX) Pretreatment and Enzymatic Hydrolysis of Miscanthus x giganteus to Fermentable Sugars
TLDR
Sugar yields during enzymatic hydrolysis from ammonia fiber expansion (AFEX) pretreated Miscanthus are reported to be 96% glucan and 81% xylan conversions, and a mass balance for the AFEX pretreatment and enzyme loadings is presented.
Enzymatic conversion of biomass for fuels production.
Bioconversion for Production of Renewable Transportation Fuels in the United States: A Strategic Perspective Cellulase and Xylanase Systems of Thermotoga neapolitana Structure-Function Studies of
Structural modification of lignocellulosics by pretreatments to enhance enzymatic hydrolysis
TLDR
The relationship between the Hydrolysis rate and the set of structural features indicated that an increase in surface area and a decrease in the crystallinity and lignin content enhance the hydrolysis; the specific surface area is the most influential of the structural features, followed by the lignIn content.
Limitations of the DNS assay for reducing sugars from saccharified lignocellulosics.
TLDR
The DNS assay can be used as an accurate analytical method for the evaluation of reducing sugars in pure solution as well as in supernatants from enzymatic saccharification if glucose is the sole product, but only specific assay methods such as HPLC and YSI-type glucose analyzers should be used for the analysis of saccharides produced from the hydrolysis of native or pretreated lignocellulosics.
...
1
2
3
4
5
...