Colonization of the cystic fibrosis lung by is greatly facilitated by the production of an exopolysaccharide called alginate. Many of the enzymes involved in alginate biosynthesis are clustered in an operon at 34 min on the chromosome. This paper reports the nucleotide sequence of a previously uncharacterized gene, which lies between the and genes of the operon. DNA sequencing data for predicted a protein product of approximately 52.5 kDa which contains a putative 27 amino acid N-terminal signal sequence and a consensus cleavage and lipid attachment site for signal peptidase II. Expression of using either T7 or promoter expression systems, and labelling studies with [S]methionine, indicated that encodes a polypeptide of approximately 53 kDa which is processed to a mature protein of approximately 50 kDa when expressed in or in agreement with the nucleotide sequence analysis. Results from an AlgK-β-lactamase fusion survey support this interpretation and also provide evidence that mature AlgK is entirely periplasmic and is probably membrane-anchored.


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