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Improvement of mannitol production by Lactobacillus brevis mutant 3-A5 based on dual-stage pH control and fed-batch fermentations

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Lactobacillus brevis 3-A5 was isolated and expected to produce mannitol efficiently by regulating pH in batch and fed-batch fermentations. In 48 h batch fermentations with free and constant pH, the optimal pH for cell growth and mannitol production in the first 24 h of incubation was 5.5, whereas that for mannitol production in the second 24 h of incubation was 4.5. To achieve high cell density and mannitol yield simultaneously, a dual-stage pH control strategy was proposed based on the kinetic analysis of mannitol production. The pH value was controlled at 5.5 for the first 12 h of fermentation and subsequently shifted to 4.5 until the fermentation was completed. Under dual-stage pH control fermentation, a 103 g/L yield of mannitol with a volumetric production rate of 3.7 g/L/h was achieved after 28 h. The dual-stage pH control fed-batch fermentation strategy was further developed to improve mannitol yield, wherein the yield increased by 109 % to 215 g/L after 98 h of fermentation. This value is the highest yield of mannitol ever reported using L. brevis.

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Fig. 1
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Fig. 3


X :

Cell dry weight, g/L

X m :

Maximum cell dry weight, g/L

μ :

Specific cell growth rate, 1/h

μ max :

Maximum specific cell growth rate, 1/h

t :

Time, h

P :

Product concentration, g/L

Q P :

Volumetric productivity of mannitol, g/L/h

α :

Growth associated constant for product formation

β :

Non-growth associated constant for product formation, 1/h

q P :

Specific mannitol formation rate, 1/h

q L :

Specific lactic acid formation rate, 1/h

q A :

Specific acetic acid formation rate, 1/h

q E :

Specific ethanol formation rate, 1/h

P/(L + A + E):

Ratio of mannitol to organic acids (lactic acid and acetic acid) and ethanol

S :

Substrate concentration, g/L

Y X/S :

Cell yield coefficient on substrate, g biomass/g sugar

Y P/X :

Product yield coefficient on biomass, g mannitol/g biomass

Y P/S :

Product yield coefficient on substrate, g mannitol/g sugar

m s :

Energy maintain constant, 1/h

q SF :

Specific fructose consumption rate, 1/h

q SG :

Specific glucose consumption rate, 1/h


Specific mannitol dehydrogenase activity on the biomass, U/mg protein/(g/L biomass)


Ratio of NADH to NAD


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This research was financially supported by the Main Direction Program of Knowledge Innovation of the Chinese Academy of Science (Grant No. KSCX2-EW-G-5), the National High Technology Research and Development Program of China (863 Program) (Grant No. 2012AA021205) and National Science and Technology Support Program (Grant No. 2012BAD32B09). We thank Dr. Haidong Tan for critical reading of the manuscript.

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Correspondence to Yuguang Du.

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Yue, M., Cao, H., Zhang, J. et al. Improvement of mannitol production by Lactobacillus brevis mutant 3-A5 based on dual-stage pH control and fed-batch fermentations. World J Microbiol Biotechnol 29, 1923–1930 (2013). https://doi.org/10.1007/s11274-013-1357-6

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  • Mannitol
  • Dual-stage pH control
  • Lactobacillus brevis
  • Batch fermentation
  • Fed-batch fermentation