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Microbiology 148 (2002), 3467-3476
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Microbiology (2002), 148, 3467-3476.
© 2002 Society for General Microbiology

Research Paper

Catabolism of mannitol in Lactococcus lactis MG1363 and a mutant defective in lactate dehydrogenase

Ana Rute Neves1, Ana Ramos1, Claire Shearman2, Michael J. Gasson2 and Helena Santos1

Instituto de Tecnologia Química e Biológica/Universidade Nova de Lisboa, and Instituto de Biologia Experimental e Tecnológica, Rua da Quinta Grande, 6, Apt 127, 2780-156 Oeiras, Portugal1
Institute of Food Research, Norwich Laboratory, Norwich Research Park, Colney, Norwich NR4 7UA, UK2

Author for correspondence: Helena Santos. Tel: +351 21 4469829. Fax: +351 21 4428766. e-mail: santos{at}itqb.unl.pt

Mannitol metabolism in Lactococcus lactis MG1363 and in a derivative strain deficient in lactate dehydrogenase (LDHd) was characterized. Both strains had the ability to grow on mannitol as an energy source, although this polyol was a poorer substrate for growth than glucose. When compared to glucose, the metabolism of mannitol caused an NADH burden due to formation of an additional NADH molecule at the reaction catalysed by mannitol-1-phosphate dehydrogenase (Mtl1PDH). This resulted in a prominent accumulation of mannitol 1-phosphate (Mtl1P) both in growing and resting cells, suggesting the existence of a severe bottleneck at Mtl1PDH. Growth on mannitol induced the activity of Mtl1PDH in both the LDHd and MG1363 strains. The lower accumulation of Mtl1P in mannitol-grown cells when compared to glucose-grown LDHd cells, as monitored by in vivo 13C-NMR, reflects this induction. A clear shift towards the production of ethanol was observed on mannitol, indicating pressure to regenerate NAD+ when this substrate was used. A strategy to obtain a mannitol-overproducing strain is proposed.

Keywords: mannitol catabolism, L. lactis, in vivo 13C-NMR

Abbreviations: FBP, fructose 1,6-bisphosphate; F6P, fructose 6-phosphate; GAPDH, glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12); LDH, L-lactate dehydrogenase (EC 1.1.1.27); LDHd, lactate-dehydrogenase-deficient; Mtl1P, mannitol 1-phosphate; Mtl1PDH, mannitol-1-phosphate dehydrogenase (EC 1.1.1.17); PEP, phosphoenolpyruvate; PFK, 6-phosphofructokinase (EC 2.7.1.11); 3-PGA, 3-phosphoglycerate; PK, pyruvate kinase (EC 2.7.1.40); PTSMtl, mannitol phosphotransferase system




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