Previous Article | Next Article 
Applied and Environmental Microbiology, September 2000, p. 3835-3841, Vol. 66, No. 9
Agricultural Research Centre of Finland, Food
Research Institute, FIN-31600 Jokioinen,1
Department of Basic Veterinary Sciences, Section of
Microbiology, FIN-00014 University of Helsinki,2
and Laboratory of Bioprocess Engineering, Helsinki
University of Technology, FIN-02015 HUT,3
Finland
Received 17 March 2000/Accepted 6 June 2000
Expression of D-(
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Metabolic Engineering of Lactobacillus
helveticus CNRZ32 for Production of Pure
L-(+)-Lactic Acid
)-lactate dehydrogenase
(D-LDH) and L-(+)-LDH genes (ldhD
and ldhL, respectively) and production of
D-()- and L-(+)-lactic acid were studied in
Lactobacillus helveticus CNRZ32. In order to develop a host
for production of pure L-(+)-isomer of lactic acid, two
ldhD-negative L. helveticus CNRZ32 strains were
constructed using gene replacement. One of the strains was constructed
by deleting the promoter region of the ldhD gene, and the
other was constructed by replacing the structural gene of
ldhD with an additional copy of the structural gene
(ldhL) of L-LDH of the same species. The
resulting strains were designated GRL86 and GRL89, respectively. In
strain GRL89, the second copy of the ldhL structural gene
was expressed under the ldhD promoter. The two
D-LDH-negative strains produced only
L-(+)-lactic acid in an amount equal to the total lactate
produced by the wild type. The maximum L-LDH activity was
found to be 53 and 93% higher in GRL86 and GRL89, respectively, than
in the wild-type strain. Furthermore, process variables for
L-(+)-lactic acid production by GRL89 were optimized using
statistical experimental design and response surface methodology. The
temperature and pH optima were 41°C and pH 5.9. At low pH, when the
growth and lactic acid production are uncoupled, strain GRL89 produced
approximately 20% more lactic acid than GRL86.
*
Corresponding author. Mailing address: Faculty of
Veterinary Medicine, Department of Basic Veterinary Sciences, Section
of Microbiology, P.O. Box 57, FIN-00014 University of Helsinki,
Finland. Phone: 358 9 191 49531. Fax: 358 9 191 49799. E-mail:
Airi.Palva{at}helsinki.fi.
Applied and Environmental Microbiology, September 2000, p. 3835-3841, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Helanto, M., Kiviharju, K., Leisola, M., Nyyssola, A.
(2007). Metabolic Engineering of Lactobacillus plantarum for Production of L-Ribulose. Appl. Environ. Microbiol.
73: 7083-7091
[Abstract] [Full Text] -
Saitoh, S., Ishida, N., Onishi, T., Tokuhiro, K., Nagamori, E., Kitamoto, K., Takahashi, H.
(2005). Genetically Engineered Wine Yeast Produces a High Concentration of L-Lactic Acid of Extremely High Optical Purity. Appl. Environ. Microbiol.
71: 2789-2792
[Abstract] [Full Text] -
Lindholm, A., Smeds, A., Palva, A.
(2004). Receptor Binding Domain of Escherichia coli F18 Fimbrial Adhesin FedF Can Be both Efficiently Secreted and Surface Displayed in a Functional Form in Lactococcus lactis. Appl. Environ. Microbiol.
70: 2061-2071
[Abstract] [Full Text] -
Juarez Tomas, M. S., Ocana, V. S., Wiese, B., Nader-Macias, M. E.
(2003). Growth and lactic acid production by vaginal Lactobacillus acidophilus CRL 1259, and inhibition of uropathogenic Escherichia coli. J Med Microbiol
52: 1117-1124
[Abstract] [Full Text] -
Avall-Jaaskelainen, S., Lindholm, A., Palva, A.
(2003). Surface Display of the Receptor-Binding Region of the Lactobacillusbrevis S-Layer Protein in Lactococcuslactis Provides Nonadhesive Lactococci with the Ability To Adhere to Intestinal Epithelial Cells. Appl. Environ. Microbiol.
69: 2230-2236
[Abstract] [Full Text] -
Zhou, S., Shanmugam, K. T., Ingram, L. O.
(2003). Functional Replacement of the Escherichia coliD-(-)-Lactate Dehydrogenase Gene (ldhA) with the L-(+)-Lactate Dehydrogenase Gene (ldhL) from Pediococcus acidilactici. Appl. Environ. Microbiol.
69: 2237-2244
[Abstract] [Full Text] -
Smeds, A., Pertovaara, M., Timonen, T., Pohjanvirta, T., Pelkonen, S., Palva, A.
(2003). Mapping the Binding Domain of the F18 Fimbrial Adhesin. Infect. Immun.
71: 2163-2172
[Abstract] [Full Text] -
Zhou, S., Causey, T. B., Hasona, A., Shanmugam, K. T., Ingram, L. O.
(2003). Production of Optically Pure D-Lactic Acid in Mineral Salts Medium by Metabolically Engineered Escherichia coli W3110. Appl. Environ. Microbiol.
69: 399-407
[Abstract] [Full Text] -
Jakava-Viljanen, M., Avall-Jaaskelainen, S., Messner, P., Sleytr, U. B., Palva, A.
(2002). Isolation of Three New Surface Layer Protein Genes (slp) from Lactobacillus brevis ATCC 14869 and Characterization of the Change in Their Expression under Aerated and Anaerobic Conditions. J. Bacteriol.
184: 6786-6795
[Abstract] [Full Text] -
Avall-Jaaskelainen, S., Kyla-Nikkila, K., Kahala, M., Miikkulainen-Lahti, T., Palva, A.
(2002). Surface Display of Foreign Epitopes on the Lactobacillus brevis S-Layer. Appl. Environ. Microbiol.
68: 5943-5951
[Abstract] [Full Text] -
Underwood, S. A., Zhou, S., Causey, T. B., Yomano, L. P., Shanmugam, K. T., Ingram, L. O.
(2002). Genetic Changes To Optimize Carbon Partitioning between Ethanol and Biosynthesis in Ethanologenic Escherichia coli. Appl. Environ. Microbiol.
68: 6263-6272
[Abstract] [Full Text]
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»