Optimization of Culture Conditions for Enhanced Lysine Production Using Engineered Escherichia coli

Ying, Hanxiao; He, Xun; Li, Yan; Chen, Kequan; Ouyang, Pingkai

doi:10.1007/s12010-014-0820-7

Cited by 41 publications

(28 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…An OD 600 of 1.0 represented 45 mg dry weight per liter. The concentration of glucose and l -lysine was analyzed using a SBA-40C biosensor analyzer [23] (Shandong Province Academy of Sciences, Jinan, China) as previously described. The protein expression were examined by SDS-PAGE.…”

Section: Methodsmentioning

confidence: 99%

Expanding metabolic pathway for de novo biosynthesis of the chiral pharmaceutical intermediate l-pipecolic acid in Escherichia coli

et al. 2017

Self Cite

View full text Add to dashboard Cite

BackgroundThe six-carbon circular non-proteinogenic compound l-pipecolic acid is an important chiral drug intermediate with many applications in the pharmaceutical industry. In the present study, we developed a metabolically engineered strain of Escherichia coli for the overproduction of l-pipecolic acid from glucose.ResultsThe metabolic pathway from l-lysine to l-pipecolic acid was constructed initially by introducing lysine cyclodeaminase (LCD). Next, l-lysine metabolic flux from glucose was amplified by the plasmid-based overexpression of dapA, lysC, and lysA under the control of the strong trc promoter to increase the biosynthetic pool of the precursor l-lysine. Additionally, since the catalytic efficiency of the key enzyme LCD is limited by the cofactor NAD+, the intracellular pyridine nucleotide concentration was rebalanced by expressing the pntAB gene encoding the transhydrogenase, which elevated the proportion of LCD with bound NAD+ and enhanced l-pipecolic acid production significantly. Further, optimization of Fe2+ and surfactant in the fermentation process resulted in 5.33 g/L l-pipecolic acid, with a yield of 0.13 g/g of glucose via fed-batch cultivation.ConclusionsWe expanded the metabolic pathway for the synthesis of the chiral pharmaceutical intermediate l-pipecolic acid in E. coli. Using the engineered E. coli, a fast and efficient fermentative production of l-pipecolic acid was achieved. This strategy could be applied to the biosynthesis of other commercially and industrially important chiral compounds containing piperidine rings.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0666-0) contains supplementary material, which is available to authorized users.

show abstract

Section: Methodsmentioning

confidence: 99%

Expanding metabolic pathway for de novo biosynthesis of the chiral pharmaceutical intermediate l-pipecolic acid in Escherichia coli

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…The concentrations of L-lysine were analyzed by using a SBA-40 C biosensor analyzer (Shandong Province Academy of Sciences, China) [20]. During the reaction, product identity was monitored by TLC.…”

Section: Methodsmentioning

confidence: 99%

Enhanced conversion of L-lysine to L-pipecolic acid using a recombinant Escherichia coli containing lysine cyclodeaminase as whole-cell biocatalyst

Ying

Wang

et al. 2015

Journal of Molecular Catalysis B: Enzymatic

Self Cite

View full text Add to dashboard Cite

“…An OD 560 of 1.0 represented 0.42 g of dry weight per liter. The broth samples were centrifuged at 32009g for 10 min (AvantiJ-26xp, Beckman, USA), and then the concentrations of L-lysine were determined using an SBA-40C biosensor analyzer (Shandong Province Academy of Sciences, China) [3,15].…”

Section: Analytical Techniquesmentioning

confidence: 99%

“…such as Corynebacterium glutamicum [2], Escherichia spp. such as Escherichia coli [3]. Nevertheless, with the continuous expansion of the global market for L-lysine, less expensive fermentation substrates are needed to reduce the costs of production [4].…”

Section: Introductionmentioning

confidence: 99%

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

Chen

et al. 2015

Bioprocess Biosyst Eng

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Optimization of Culture Conditions for Enhanced Lysine Production Using Engineered Escherichia coli

Cited by 41 publications

References 20 publications

Expanding metabolic pathway for de novo biosynthesis of the chiral pharmaceutical intermediate l-pipecolic acid in Escherichia coli

Expanding metabolic pathway for de novo biosynthesis of the chiral pharmaceutical intermediate l-pipecolic acid in Escherichia coli

Enhanced conversion of L-lysine to L-pipecolic acid using a recombinant Escherichia coli containing lysine cyclodeaminase as whole-cell biocatalyst

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

Contact Info

Product

Resources

About