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Enhanced l-lysine production from pretreated beet molasses by engineered Escherichia coli in fed-batch fermentation

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Abstract

Faster sugar consumption rate and low-cost nitrogen source are required for the chemical biosynthesis using molasses. Five pretreatment methods were applied to beet molasses prior to fermentation through engineered Escherichia coli, respectively, and corn steep liquid was used as an organic nitrogen source to replace expensive yeast extract. Furthermore, the effects of different feeding strategy in fed-batch fermentation on l-lysine production were investigated. The experimental results showed that combined tricalcium phosphate, sulfuric acid, and activated carbon pretreatment method (TPSA) pretreatment could improve the sugar consumption rate most greatly, and the initial total sugar concentration of 35 g/L from TPSA-pretreated beet molasses gave the best results with respect to l-lysine production, dry cell weight concentration, and l-lysine yield in batch fermentation. Moreover, a mixture of low-cost corn steep liquid and yeast extract containing equal amount of nitrogen could be used as the organic nitrogen source for effective l-lysine fermentation, and constant speed feeding strategy of TPSA-pretreated beet molasses promoted l-lysine production by engineered E. coli. The TPSA-pretreated beet molasses had a sugar consumption rate of 1.75 g/(L h), and a l-lysine yield of 27.81 % was achieved, compared with the theoretical yield of 62 % by glucose. It was clarified that the pretreatment significantly enhanced the conversion of sugars in beet molasses to l-lysine.

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Abbreviations

MF:

Microfiltration pretreatment method

AC:

Activated carbon pretreatment method

SA:

Sulfuric acid pretreatment method

TCPH:

Tricalcium phosphate and sulfuric acid pretreatment method

TPSA:

Combined tricalcium phosphate, sulfuric acid, and activated carbon pretreatment method

CSL:

Corn steep liquor powder

TN:

Total nitrogen, g/g

DCW:

E. coli dry cell weight, g/L

OD560 :

Optical density measured at 560 nm

l-Lysine yield:

The final amount of l-lysine produced from 1 g of glucose, and expressed as a mass ratio

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Acknowledgments

This work was supported by the National Nature Science Foundation of China (Grant nos. 21390200, 21106068), National Key Technology Support Program (Grant no. 2012BAI44G00), “973” program of China (Grant no. 2011CBA00807), and “863” program of China (Grant no. 2014AA021703).

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Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical, Nanjing Tech University, Nanjing, 211816, People’s Republic of China

    Xun He, Kequan Chen, Yan Li, Zhen Wang, Hong Zhang, Juan Qian & Pingkai Ouyang

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  1. Xun He
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  2. Kequan Chen
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  3. Yan Li
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  4. Zhen Wang
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  5. Hong Zhang
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  6. Juan Qian
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  7. Pingkai Ouyang
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Correspondence to Kequan Chen.

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He, X., Chen, K., Li, Y. et al. Enhanced l-lysine production from pretreated beet molasses by engineered Escherichia coli in fed-batch fermentation. Bioprocess Biosyst Eng 38, 1615–1622 (2015). https://doi.org/10.1007/s00449-015-1403-x

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  • DOI: https://doi.org/10.1007/s00449-015-1403-x

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