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

Neves, Ana Rute; Ramos, António Manuel Pinho; Shearman, Claire; Gasson, Michael J.; Santos, Helena

doi:10.1099/00221287-148-11-3467

Cited by 36 publications

(41 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…M1Pase activity in this strain was induced by the addition of various concentrations of nisin. The endogeneous M1PDH was induced in this strain by preculturing cells on M17 broth containing mannitol, since the mannitol utilization genes, including the mtlD gene encoding M1PDH, are activated during growth on mannitol (23,30). Induction with increasing levels of nisin in these mannitol-induced cells during growth on glucose resulted in a stable and high level of M1PDH activity, and an increasing level of M1Pase activity (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The results shown here emphasize the importance of M1Pase activity for mannitol production by L. lactis and indicate that a L. lactis strain deficient in LDH activity and with high M1PDH and M1Pase activity would be a good candidate for in situ mannitol production in food products. Regarding the possibility of the use of such a L. lactis in a microbial mannitol production process, an advantage of mannitol production by a L. lactis strain in comparison with heterofermentative LABs such as Leuconostoc mesenteroides (13,28) might be the capability of L. lactis to use several sugar substrates to synthesize mannitol, whereas mannitol is exclusively formed from fructose by the (23,30). Therefore, it is not unlikely that reutilization of mannitol would occur eventually after repeated subculturing, due to induction of the mannitol transport system by mannitol (22,23).…”

Section: Discussionmentioning

confidence: 99%

“…Regarding the possibility of the use of such a L. lactis in a microbial mannitol production process, an advantage of mannitol production by a L. lactis strain in comparison with heterofermentative LABs such as Leuconostoc mesenteroides (13,28) might be the capability of L. lactis to use several sugar substrates to synthesize mannitol, whereas mannitol is exclusively formed from fructose by the (23,30). Therefore, it is not unlikely that reutilization of mannitol would occur eventually after repeated subculturing, due to induction of the mannitol transport system by mannitol (22,23). Inactivation of such a mannitol transport system, as recently published (12), could prevent the reutilization of mannitol in strains overproducing the large amounts of mannitol as presented here.…”

Section: Discussionmentioning

confidence: 99%

“…To create a mannitol-producing L. lactis strain, several metabolic engineering strategies have been applied, such as inactivation of LDH activity and a mannitol transport system (12,22,23). With L. lactis and L. plantarum, inactivation of NADH consuming pathways such as LDH has been proven to be a successful strategy to invoke alternative NAD ϩ regeneration through mannitol production (11,12,22).…”

Section: Discussionmentioning

confidence: 99%

“…Transient formation of high concentrations of intracellular mannitol (90 mM) and mannitol 1-phosphate (76 mM) were detected in high-density nongrowing cell suspensions of a lactate dehydrogenase (LDH)-deficient L. lactis strain (22). During growth, only small amounts of mannitol (Ͻ0.4 mM) were transiently produced extracellularly (23). Recently, inactivation of the mannitol transport system in an LDH-deficient L. lactis strain resulted in high extracellular mannitol production.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Overproduction of Heterologous Mannitol 1-Phosphatase: a Key Factor for Engineering Mannitol Production byLactococcus lactis

Wisselink

Moers

Mars

et al. 2005

Appl Environ Microbiol

View full text Add to dashboard Cite

To achieve high mannitol production by Lactococcus lactis, the mannitol 1-phosphatase gene of Eimeria tenella and the mannitol 1-phosphate dehydrogenase gene mtlD of Lactobacillus plantarum were cloned in the nisindependent L. lactis NICE overexpression system. As predicted by a kinetic L. lactis glycolysis model, increase in mannitol 1-phosphate dehydrogenase and mannitol 1-phosphatase activities resulted in increased mannitol production. Overexpression of both genes in growing cells resulted in glucose-mannitol conversions of 11, 21, and 27% by the L. lactis parental strain, a strain with reduced phosphofructokinase activity, and a lactate dehydrogenasedeficient strain, respectively. Improved induction conditions and increased substrate concentrations resulted in an even higher glucose-to-mannitol conversion of 50% by the lactate dehydrogenase-deficient L. lactis strain, close to the theoretical mannitol yield of 67%. Moreover, a clear correlation between mannitol 1-phosphatase activity and mannitol production was shown, demonstrating the usefulness of this metabolic engineering approach.Mannitol is a reduced form of fructose and is produced by a variety of microorganisms including bacteria, yeasts, and fungi. Besides the ability of several organisms to maintain their redox balance by the production of mannitol (9, 21, 22), mannitol has a physiological function in microorganisms as an osmolyte (16) and can serve as a protecting agent. It has been reported that mannitol enhances survival of Lactococcus lactis cells during drying processes (10). The viability of starter cultures of L. lactis, a lactic acid bacterium (LAB) extensively used in dairy industry, may thus be enhanced by mannitol production. In addition, the use of a mannitol-producing L. lactis may result in fermented products with extra value, since mannitol is assumed to have several beneficial effects as a food additive. It can serve as an antioxidant (4,5,25,26) and as a low-calorie sweetener that can replace sucrose (6,8).In heterofermentative LABs such as Leuconostoc mesenteroides, mannitol is formed from fructose in a single conversion by mannitol dehydrogenase, and fructose-to-mannitol conversion rates of Ͼ90% are common (13,24,27). In contrast, most homofermentative LABs, such as Lactococcus lactis, do not normally produce mannitol. Mannitol formation in homofermentative LABs is limited to strains whose ability to regenerate NAD to fulfill the redox balance is severely hampered. In these strains, mannitol 1-phosphate dehydrogenase (M1PDH) and mannitol 1-phosphatase (M1Pase) are the enzymes involved in the mannitol biosynthesis route (Fig. 1). Transient formation of high concentrations of intracellular mannitol (90 mM) and mannitol 1-phosphate (76 mM) were detected in high-density nongrowing cell suspensions of a lactate dehydrogenase (LDH)-deficient L. lactis strain (22). During growth, only small amounts of mannitol (Ͻ0.4 mM) were transiently produced extracellularly (23). Recently, inactivation of the mannitol transport system in an LDH-def...

show abstract

Section: Resultsmentioning

confidence: 99%