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Metabolic Engineering of Acetaldehyde Production by Streptococcus thermophilus

A. C. S. D. Chaves,^1,2 M. Fernandez,^1, A. L. S. Lerayer,³ I. Mierau,¹ M. Kleerebezem,¹ and J. Hugenholtz^1*

NIZO Food Research and Wageningen Centre for Food Sciences (WCFS), Ede, The Netherlands,¹ State University of Campinas (UNICAMP),² Instituto de Tecnologia de Alimentos (ITAL), Campinas SP, Brazil³

Received 25 April 2002/ Accepted 20 August 2002

The process of acetaldehyde formation by the yogurt bacterium Streptococcus thermophilus is described in this paper. Attention was focused on one specific reaction for acetaldehyde formation catalyzed by serine hydroxymethyltransferase (SHMT), encoded by the glyA gene. In S. thermophilus, SHMT also possesses threonine aldolase (TA) activity, the interconversion of threonine into glycine and acetaldehyde. In this work, several wild-type S. thermophilus strains were screened for acetaldehyde production in the presence and absence of L-threonine. Supplementation of the growth medium with L-threonine led to an increase in acetaldehyde production. Furthermore, acetaldehyde formation during fermentation could be correlated to the TA activity of SHMT. To study the physiological role of SHMT, a glyA mutant was constructed by gene disruption. Inactivation of glyA resulted in a severe reduction in TA activity and complete loss of acetaldehyde formation during fermentation. Subsequently, an S. thermophilus strain was constructed in which the glyA gene was cloned under the control of a strong promoter (P_LacA). When this strain was used for fermentation, an increase in TA activity and in acetaldehyde and folic acid production was observed. These results show that, in S. thermophilus, SHMT, displaying TA activity, constitutes the main pathway for acetaldehyde formation under our experimental conditions. These findings can be used to control and improve acetaldehyde production in fermented (dairy) products with S. thermophilus as starter culture.

Present address: Instituto de Productos Lacteos de Asturias (CSIC), Villaviciosa, Asturias, Spain.

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