The Johns Hopkins University

New England Biolabs, Inc.

Saul Roseman

Charles Y Yu

Edith Chitlaru

Nemat O Keyhani

Bonnie Bassler

Donald G Comb

Xibing Li

Chris Rowe

Compositions and methods are provided for converting chitin into N-acetylglucosamine, glucosamine and ethanol. The chitin may be used directly from the environment, for example, as occurs in invertebrate cuticles, fungal cells and/or algae. Mutant bacteria were created by knocking out or inactivating one or more genes preferably resulting in the chitin catabolic sensor maintaining an activated state. Methods are further provided for converting the N-acetylglucosamine into ethanol by means of a genetically engineered yeast strain which can be optionally co-cultivated with the Vibrionaceae to produce significant yields of ethanol.

Conversion of chitin into N-acetylglucosamine, glucosamine and bioethanol

Three genes involved in the catabolism of chitin in Vibrio furnissii: endI encodes periplasmic chitodextrinase, exoI encodes periplasmic .beta.-N-acetylglucosaminidase, and exoII encodes aryl .beta.-N-acetylglucosaminidase are provided. The complete nucleotide sequence for each of the three genes and the complete amino acid for the corresponding enzymes are demonstrated along with host cells capable of expressing the recombinant enzymes. The present invention also describes four specific strains of V. furnissii having deletions in genes involved in the catabolic pathway of chitin and a process for the production of chitin oligosaccharides.

Bacterial catabolism of chitin

The present invention reveals the cloning of four genes involved in the catabolism of chitin in Vibrio furnissii: endI encodes periplasmic chitodextrinase, exoI encodes periplasmic .beta.-N-acetylglucosaminidase, exoII encodes aryl .beta.-N-acetyl-glucosaminidase and chiA encodes extracellular chitinase. The complete nucleotide sequence for each of the four genes and the complete amino acid for the corresponding enzymes are demonstrated along with host cells capable of expressing the recombinant enzymes. The present invention also describes four specific strains of V. furnissii having deletions in genes involved in the catabolic pathway of chitin and a process for the production of chitin oligosaccharides.

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Baltimore, MD, United States of America

Saul Roseman