KAIROS Scientific Awarded $500K for Enzyme Development for Cellulosic Ethanol and Other Applications
4 October 2005
KAIROS Scientific , a biotechnology company in the fields of digital imaging spectroscopy and protein engineering, announced that it has been awarded a $500,000 grant from the National Science Foundation (NSF).
The Phase II Small Business Innovation Research (SBIR) grant will be used to accelerate the development of optimized cellulases—enzymes that selectively degrade cellulose fibers derived from wood, plant material and recycled paper.
KAIROS will employ its patented Kcat technology for directed evolution of the enzymes. Directed evolution is a general term that describes a set of molecular biology techniques that mimic natural selection for the iterative production, evaluation and selection of variants of a biological sequence.
In essence, bioengineers make modifications to genes, introduce those genes into many microcolonies of bacteria, screen the results, pick the offspring with the most desirable traits (expression of an enzyme in this case), and repeat the process again.
The enzymes will be designed for greater stability and resistance to common inhibitors, thus making them more economically viable for use in industry. The engineered cellulases may be applied to papermaking, recycling and the efficient conversion of agricultural waste into ethanol for fuel.
By carefully controlling in vitro mutation efficiencies and screening for enhanced catalytic properties over multiple generations, researchers have developed new enzymes that are hundreds of times more active than the natural enzymes in chemical process environments.
The bottleneck in the process is the time it takes to screen through all the outcomes. That’s where KAIROS’ Kcat technology comes in.
KCat is a high-throughput solid-phase assay system consisting of a new digital imaging spectrophotometer and a series of colorimetric solid phase assays for screening bacteria that are expressing mutagenized enzymes undergoing directed evolution.
The system can detect less than a 20% difference in enzyme rates within microcolonies grown at a nearly confluent density of 500 colonies per cm2. Each microcolony is analyzed simultaneously at single-pixel resolution (1.5 megapixels; 75 micron/pixel), requiring less than 100 nanoliters of substrate per measurement, a 1000-fold reduction over conventional liquid phase assays.
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