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Journal of Bacteriology, August 1999, p. 4592-4597, Vol. 181, No. 15
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Genetic Characterization of a Cell
Envelope-Associated Proteinase from Lactobacillus
helveticus CNRZ32
Jeffrey A.
Pederson,1
Gerald J.
Mileski,2,3
Bart
C.
Weimer,2,3 and
James L.
Steele1,*
Department of Food Science, University of
Wisconsin
Madison, Madison, Wisconsin 53706,1
and Western Center for Dairy Research2
and Department of Nutrition and Food
Sciences,3 Utah State University, Logan, Utah
84322
Received 16 March 1999/Accepted 21 May 1999
A cell envelope-associated proteinase gene (prtH) was
identified in Lactobacillus helveticus CNRZ32. The
prtH gene encodes a protein of 1,849 amino acids and with a
predicted molecular mass of 204 kDa. The deduced amino acid sequence of
the prtH product has significant identity (45%) to that of
the lactococcal PrtP proteinases. Southern blot analysis indicates that
prtH is not broadly distributed within L. helveticus. A prtH deletion mutant of CNRZ32 was
constructed to evaluate the physiological role of PrtH. PrtH is not
required for rapid growth or fast acid production in milk by CNRZ32.
Cell surface proteinase activity and specificity were determined by
hydrolysis of
s1-casein fragment 1-23 by whole cells. A
comparison of CNRZ32 and its prtH deletion mutant indicates that CNRZ32 has at least two cell surface proteinases that differ in
substrate specificity.
*
Corresponding author. Mailing address: Department of
Food Science, University of Wisconsin
Madison, 1605 Linden Dr.,
Madison, WI 53706-1565. Phone: (608) 262-5960. Fax: (608) 262-6872. E-mail: jlsteele{at}facstaff.wisc.edu.
Journal of Bacteriology, August 1999, p. 4592-4597, Vol. 181, No. 15
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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