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An Algorithmically Optimized Combinatorial Library Screened by Digital Imaging Spectroscopy

Bio/Technology volume 10, pages 1557–1561 (1992)Cite this article

Abstract

Combinatorial cassettes based on a phylogenetic “target set” were used to simultaneously mutagenize seven amino acid residues on one face of a transmembrane alpha helix comprising a bacteriochlorophyll binding site in the light harvesting n antenna of Rhodobacter capsulatus. This pigmented protein provides a model system for developing complex mutagenesis schemes, because simple absorption spectroscopy can be used to assay protein expression, structure, and function. Colony screening by Digital Imaging Spec-troscopy showed that 6% of the optimized library bound bacteriochlorophyll in two distinct spectroscopic classes. This is approximately 200 times the throughput (ca. 0.03%) of conventional combinatorial cassette mutagenesis using [NN(G/C)]7. “Doping” algorithms evaluated in this model system are generally applicable and should enable simultaneous mutagenesis at more positions in a protein than currently possible, or alternatively, decrease the screening size of combinatorial libraries.

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  1. Douglas C. Youvan: Corresponding author.

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  1. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Ellen R. Goldman & Douglas C. Youvan

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  1. Ellen R. Goldman
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  2. Douglas C. Youvan
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Goldman, E., Youvan, D. An Algorithmically Optimized Combinatorial Library Screened by Digital Imaging Spectroscopy. Nat Biotechnol 10, 1557–1561 (1992). https://doi.org/10.1038/nbt1292-1557

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