Kcat Technology from KAIROS is an advanced assay system based on Digital Imaging Spectroscopy (DIS) that is designed for high-throughput screening of enzyme variants. Using Directed Evolution and Kcat Technology, millions of genetic variants of your enzyme can be screened for:

• thermal stability
• tolerance to extreme pH
• modified substrate specificity
• better catalysis
• improved activity in nonaqueous solvents
• enhanced enantioselectivity

Kcat combines microcolony assays in a solid-phase format with instrumentation and algorithms to acquire spectral and kinetics measurements of large expression libraries. Massively parallel data acquisition permits fast, accurate assays using only nanoliter amounts of substrate per variant. The results: vastly higher throughput than conventional formats, greater assay flexibility (including activity measurements on high molecular weight substrates), and lower cost per assay.  

Custom enzymes whenever you want them. Kcat technology is so user-friendly that it makes it possible for virtually any laboratory to produce its own custom enzymes for any desired purpose. KAIROS will help you set up your own in-house screening facility. Alternatively, we can screen libraries for you, using our extensive expertise in mutagenesis, library construction, protein engineering and directed evolution.

Ultra-high throughput library screening. Kcat technology revolutionizes traditional chromogenic and fluorogenic enzyme assays by giving the user the power to perform them in a massively parallel format. It’s like operating over 100,000 spectrophotometers simultaneously for each assay. Each instrument is capable of screening over 1,000,000 enzyme variants per day.

Vastly simplified handling and dispensing. Unlike most liquid-phase formats, Kcat solid-phase technology requires almost no pipetting. This feature obviates the need for large, expensive, complex robotics, and makes it simple to handle viscous or insoluble substrates.

Wide substrate range. Kcat is ideally suited for high molecular weight substrates such as DNA, RNA, proteins, and polysaccharides, as well as polymers and solid materials. It is also ideal for substrates that cannot penetrate intact cells and for coupled enzyme assays.

Greatly reduced substrate requirements. Each microcolony requires less than 50 nanoliters of substrate volume per assay, greatly reducing the cost of screening large libraries on expensive substrates.

Simultaneous screening of multiple substrates. Kcat technology is designed to perform highly sophisticated enzyme assays, with full kinetics and spectra for every variant, not just endpoints or "one-shot" fluorescence measurements. This makes it possible to find not only the most active variants, but also the variants with improved regio- or enantioselectivity or altered substrate specificity.

Screen for multiple properties simultaneously. The solid-phase format makes it easy and convenient to expose the microcolonies to a variety of conditions, including elevated temperature, extremes of pH, chemical inhibitors, and non-aqueous solvents.

The Kcat instrument is self-contained and easy to use.   The instrument comes with HyperDIS software for instrument control, data acquisition, analysis, and display.  Numerous image processing algorithms are included as well as proprietary sorting algorithms to rapidly and concisely identify desirable enzyme variants out of the tens of thousands of variants screened.  

Flexible licensing and leasing arrangements.  Contact us about arrangements to help you customize an enzyme to your specifications.

See an on-line publication of Kcat Technology

Solid Phase Enzyme Screening.  ASM News, 2000.

Application of a very high-throughput digital imaging screen to evolve the enzyme galactose oxidase.  Protein Engineering, 2001

KCAT^TM patents are issued and pending.

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