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Journal of Bacteriology, April 2007, p. 3072-3080, Vol. 189, No. 8
0021-9193/07/$08.00+0     doi:10.1128/JB.01772-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Lipolytic Enzymes in Myxococcus xanthus{triangledown}

Aurelio Moraleda-Muñoz and Lawrence J. Shimkets*

Department of Microbiology, University of Georgia, Athens, Georgia 30602

Received 21 November 2006/ Accepted 5 February 2007

The genome of Myxococcus xanthus encodes lipolytic enzymes in three different families: patatin lipases, {alpha}/ß hydrolases, and GDSL lipases. One member of each family was characterized. The protein encoded by MXAN_3852 contains motifs characteristic of patatins. MXAN_5522 encodes a protein with the G-X-S-X-G motif characteristic of the lipase subfamily of {alpha} hydrolases. MXAN_4569 encodes a member of the GDSL family of lipolytic enzymes. Strains with deletions of MXAN_5522 and MXAN_4569 undergo faster development and earlier myxospore formation than the wild-type strain. The MXAN_5522 mutation results in spore yields substantially higher than those seen for wild-type cells. Gene expression analysis using translational lacZ fusions indicates that while all three genes are expressed during development, only MXAN_5522 and MXAN_4569 are expressed during vegetative growth. The proteins encoded by these genes were overexpressed using a T7 RNA polymerase transcription (pET102/D-TOPO) system in Escherichia coli BL21 Star (DE3) cells. The substrate specificities of the purified enzymes were investigated using p-nitrophenyl esters with chain lengths from C2 to C16. These enzymes preferentially hydrolyzed esters of short-chain fatty acids, yielding the highest activity with p-nitrophenyl acetate.


* Corresponding author. Mailing address: Department of Microbiology, University of Georgia, Athens, GA 30602. Phone: (706) 542-2681. Fax: (706) 542-2674. E-mail: shimkets{at}uga.edu

{triangledown} Published ahead of print on 16 February 2007.


Journal of Bacteriology, April 2007, p. 3072-3080, Vol. 189, No. 8
0021-9193/07/$08.00+0     doi:10.1128/JB.01772-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.