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Applied and Environmental Microbiology, September 1999, p. 4211-4215, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Replacement of a Metabolic Pathway for Large-Scale Production of Lactic Acid from Engineered Yeasts

Danilo Porro,1,* Michele M. Bianchi,2 Luca Brambilla,1 Rossella Menghini,2 Davide Bolzani,3 Vittorio Carrera,3 Jefferson Lievense,4 Chi-Li Liu,4 Bianca Maria Ranzi,3 Laura Frontali,2 and Lilia Alberghina1

Dipartimento di Biotecnologie e Bioscienze, Università degli Studi Milano-Bicocca, 20126 Milan,1 Dipartimento di Biologia Cellulare e dello Sviluppo, Università di Roma La Sapienza, Rome 00185;,2 and Dipartimento di Fisiologia e Biochimica Generali, Sezione Biochimica Comparata, Università degli Studi di Milano, 20133 Milan,3 Italy, and A. E. Staley Manufacturing Co., Decatur, Illinois 625214

Received 20 January 1999/Accepted 17 June 1999

Interest in the production of L-(+)-lactic acid is presently growing in relation to its applications in the synthesis of biodegradable polymer materials. With the aim of obtaining efficient production and high productivity, we introduced the bovine L-lactate dehydrogenase gene (LDH) into a wild-type Kluyveromyces lactis yeast strain. The observed lactic acid production was not satisfactory due to the continued coproduction of ethanol. A further restructuring of the cellular metabolism was obtained by introducing the LDH gene into a K. lactis strain in which the unique pyruvate decarboxylase gene had been deleted. With this modified strain, in which lactic fermentation substituted completely for the pathway leading to the production of ethanol, we obtained concentrations, productivities, and yields of lactic acid as high as 109 g liter-1, 0.91 g liter-1 h-1, and 1.19 mol per mole of glucose consumed, respectively. The organic acid was also produced at pH levels lower than those usual for bacterial processes.

* Corresponding author. Mailing address: Dipartimento di Biotecnologie e Bioscienze, University of Milan-Bicocca, Via Emanueli 12, 20126 Milano, Italy. Phone: 39 0264483451. Fax: 39 0264483233. E-mail: DANILO.PORRO{at}UNIMIB.IT.

Applied and Environmental Microbiology, September 1999, p. 4211-4215, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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