2,3-Butanediol Synthesis and the Emergence of the Vibrio cholerae El Tor Biotype

Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Ave. Boston, MA 02115

^* To whom correspondence should be addressed. Email: jmekalanos{at}hms.harvard.edu.

	Abstract

Vibrio cholerae is an aquatic bacterium that causes the severe diarrheal disease cholera. V. cholerae strains of the O1 serogroup exist as two biotypes, classical and El Tor. Toxigenic strains of the El Tor biotype emerged to cause the 7th pandemic of cholera in 1961 and subsequently displaced strains of the classical biotype both in the environment and as a cause of cholera within a decade. The factors that drove emergence of the El Tor biotype and the displacement of the classical biotype are unknown. Here, we show a unique difference in carbohydrate metabolism between these two biotypes. When grown with added carbohydrates, classical biotype strains generated a sharp decrease in media pH resulting in loss of viability. However, growth of El Tor biotype strain, N16961, was enhanced due to its ability to produce 2,3-butanediol, a neutral fermentation end-product, and suppress the accumulation of organic acids. A N16961 mutant (SSY01) defective in 2,3-butanediol synthesis showed the same defect in growth that classical biotype strains show in media rich in carbohydrates. Importantly, the SSY01 mutant was attenuated in its ability to colonize the intestine of infant mice suggesting that host carbohydrates may be available to V. cholerae within the intestinal environment. Similarly, the SSY01 mutant failed to develop biofilms when utilizing N-acetyl-D-glucosamine as a carbon source. Because, growth on N-acetyl-D-glucosamine likely reflects the ability of a strain to grow on chitin in certain aquatic environments, we conclude that the strains of classical biotype are likely defective compared to those of El Tor in growth in any environmental niche that is rich in chitin and/or other metabolizable carbohydrates. We propose that the ability to metabolize sugars without production of acid by-products might account for the improved evolutionary fitness of the V. cholerae El Tor biotype compared to the classical biotype both as a global cause of cholera and as an environmental organism.