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Applied and Environmental Microbiology, November 2001, p. 5094-5099, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5094-5099.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Characterization of Two New Glycosyl Hydrolases from the Lactic Acid Bacterium Carnobacterium piscicola Strain BA

Jonna Coombs¹ and Jean E. Brenchley^2,*

Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, New Jersey 08901-8525,¹ and Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802²

Received 22 May 2001/Accepted 3 September 2001

Three genes with homology to glycosyl hydrolases were detected on a DNA fragment cloned from a psychrophilic lactic acid bacterium isolate, Carnobacterium piscicola strain BA. A 2.2-kb region corresponding to an -galactosidase gene, agaA, was followed by two genes in the same orientation, bgaB, encoding a 2-kb -galactosidase, and bgaC, encoding a structurally distinct 1.76-kb -galactosidase. This gene arrangement had not been observed in other lactic acid bacteria, including Lactococcus lactis, for which the genome sequence is known. To determine if these sequences encoded enzymes with - and -galactosidase activities, we subcloned the genes and examined the enzyme properties. The -galactosidase, AgaA, hydrolyzes para-nitrophenyl--D-galactopyranoside and has optimal activity at 32 to 37°C. The -galactosidase, BgaC, has an optimal activity at 40°C and a half-life of 15 min at 45°C. The regulation of these enzymes was tested in C. piscicola strain BA and activity on both - and -galactoside substrates decreased for cells grown with added glucose or lactose. Instead, an increase in activity on a phosphorylated -galactoside substrate was found for the cells supplemented with lactose, suggesting that a phospho-galactosidase functions during lactose utilization. Thus, the two -galactosidases may act synergistically with the -galactosidase to degrade other polysaccharides available in the environment.

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^* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, Pennsylvania State University, 209 S. Frear, University Park, PA 16802. Phone: (814) 863-7794. Fax: (814) 865-3330. E-mail: jeb7{at}psu.edu.

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Applied and Environmental Microbiology, November 2001, p. 5094-5099, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5094-5099.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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