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Applied and Environmental Microbiology, January 2003, p. 304-311, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.304-311.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Plasmid-Encoded Diacetyl (Acetoin) Reductase in Leuconostoc pseudomesenteroides

Fergal P. Rattray,* Dorte Myling-Petersen, Dianna Larsen, and Dan Nilsson

Department of Genomics and Strain Development, Chr. Hansen A/S., DK-2970 Hørsholm, Denmark

Received 22 July 2002/ Accepted 21 October 2002

A plasmid-borne diacetyl (acetoin) reductase (butA) from Leuconostoc pseudomesenteroides CHCC2114 was sequenced and cloned. Nucleotide sequence analysis revealed an open reading frame encoding a protein of 257 amino acids which had high identity at the amino acid level to diacetyl (acetoin) reductases reported previously. Downstream of the butA gene of L. pseudomesenteroides, but coding in the opposite orientation, a putative DNA recombinase was identified. A two-step PCR approach was used to construct FPR02, a butA mutant of the wild-type strain, CHCC2114. FPR02 had significantly reduced diacetyl (acetoin) reductase activity with NADH as coenzyme, but not with NADPH as coenzyme, suggesting the presence of another diacetyl (acetoin)-reducing activity in L. pseudomesenteroides. Plasmid-curing experiments demonstrated that the butA gene is carried on a 20-kb plasmid in L. pseudomesenteroides.

* Corresponding author. Mailing address: Department of Genomics and Strain Development, Chr. Hansen A/S., 10-12 Bøge Allé, DK-2970 Hørsholm, Denmark. Phone: 45 45748545. Fax: 45 45748994. E-mail: FergalPatrick.Rattray{at}dk.chr-hansen.com.

Applied and Environmental Microbiology, January 2003, p. 304-311, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.304-311.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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