Enhanced l-phenylalanine biosynthesis by co-expression of pheAfbr and aroFwt

Zhou, Haïyan; Liao, Xianyan; Wang, Tianwen; Du, Guocheng; Chen, Jian

doi:10.1016/j.biortech.2010.01.043

Cited by 70 publications

(49 citation statements)

References 31 publications

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“…Plasmid pAP-B03 with E. coli genes aroF (encoding L-Tyr-sensitive DS) and pheA fbr (encoding L-Phe feedbackresistant CM-PDT) under the control of the temperatureinducible phage lambda promoters (pR and pL), respectively, was constructed in our previous work [29]. Kanamycin (Kan) resistance was employed as a selection marker for plasmidcontaining cells.…”

Section: Methodsmentioning

confidence: 99%

“…The cell concentration was measured as the optical density at 610 nm (OD 610 ) with a spectrophotometer-722 (Third Analytical Instrument Factory, Shanghai, China) after an appropriate dilution, where one unit of OD 610 was equivalent to 0.32 g/l dry cell weight (DCW) [29]. To determine DCW, 10 ml of fermentation broth in a pre-weighed centrifuge tube was centrifuged for 10 min at 10,000 rpm and 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…Production of L-Phe by recombinant E. coli was conducted in a fermentation medium slightly modified according to [7,29] Shake flask fermentations were carried out in 500-ml Erlenmeyer flasks containing 50 ml fermentation medium inoculated with 10% (v/v) seed cultures at 33°C and 200 rpm on rotary shakers. Calcium carbonate (12 g/l) was added to adjust the pH of the medium.…”

Section: Media and Cultivation Conditionsmentioning

confidence: 99%

“…Combining an efficient fermentation process, they achieved a final L-Phe titer of 50 g/l and a yield on glucose of 0.25 mol/mol after 36 h. This has been the highest level reported so far, however, the genetic basis of the L-Phe over-production is unrevealed. In our previous work [29], a metabolically engineered strain, E. coli WSH-Z06 (pAP-B03), was constructed by introducing a recombinant plasmid pAP-B03 carrying a novel pheA fbr gene as well as a wild-type aroF gene [encoding L-tyrosine (L-Tyr)-sensitive DS]. This strain exhibited strong resistance to high-level L-Phe and good potential for over-production of L-Phe: 35.38 g/l L-Phe with a yield coefficient of 0.26 mol/ mol glucose in pH-stat fed-batch culture within 58 h. A further improved titer of around 55 g/l at 50 h has been achieved via the optimization of fermentation conditions of L-Phe production by E. coli WSH-Z06 (pAP-B03) (unpublished data).…”

Section: Introductionmentioning

confidence: 99%

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Enhanced l-phenylalanine production by recombinant Escherichia coli BR-42 (pAP-B03) resistant to bacteriophage BP-1 via a two-stage feeding approach

Zhou

Liao

Liu

et al. 2010

J Ind Microbiol Biotechnol

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The L-phenylalanine (L-Phe) production by Escherichia coli WSH-Z06 (pAP-B03) was frequently prevented by bacteriophage BP-1 infestation. To cope with the bacteriophage BP-1 problem for an improved L-Phe production, one bacteriophage BP-1-resistant mutant, E. coli BR-42, was obtained from 416 mutant colonies of E. coli WSH-Z06 after N-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutagenesis by selection for resistance to bacteriophage BP-1. The recombinant E. coli BR-42-carrying plasmid pAP-B03 had a high capacity in L-Phe production and a remarkable tolerance to 1 × 10(10) pfu (plaque-forming unit)/ml bacteriophage stock. For an enhanced L-Phe production by E. coli BR-42 (pAP-B03), the effects of different feeding strategies including pH-stat, constant rate feeding, linear decreasing rate feeding, and exponential feeding on L-Phe production were investigated; and a two-stage feeding strategy, namely exponential feeding at μ (set) = 0.18 h(-1) in the first 20 h and a following linear varying rate feeding with F = (-0.55 × t + 18.6) ml/h, was developed to improve L-Phe production. With this two-stage feeding approach, a maximum L-Phe titer of 57.63 g/l with a high L-Phe productivity (1.15 g/l/h) was achieved, which was 15% higher than the highest level (50 g/l) reported so far according to our knowledge. The recombinant E. coli BR-42 (pAP-B03) is a potential L-Phe over-producer in substantial prevention of bacteriophage BP-1 infestation compared to its parent strain WSH-Z06 (pAP-B03).

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Media and Cultivation Conditionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Enhanced l-phenylalanine production by recombinant Escherichia coli BR-42 (pAP-B03) resistant to bacteriophage BP-1 via a two-stage feeding approach

Zhou

Liao

Liu

et al. 2010

J Ind Microbiol Biotechnol

Self Cite

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“…For example, it is a precursor of some anticancer drugs and the dipeptide sweetener aspartame [1]. In early industrial processes, L-Phe was mainly produced by chemical synthesis.…”

Section: Introductionmentioning

confidence: 99%

The influence of hydroxypropyl-β-cyclodextrin on the enantioselective hydrolysis of 2-amino phenylpropionitrile catalyzed by recombinant nitrilase

Wang

2014

Bioresour. Bioprocess.

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Enhancement of l‐phenylalanine production in Escherichia coli by heterologous expression of Vitreoscilla hemoglobin

Guo

Zhang

et al. 2017

Biotech and App Biochem

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l-Phenylalanine is an important amino acid that is widely used in the production of food flavors and pharmaceuticals. Generally, l-phenylalanine production by engineered Escherichia coli requires a high rate of oxygen supply. However, the coexpression of Vitreoscilla hemoglobin gene (vgb), driven bya tac promoter, with the genes encoding 3-deoxy-d-arabinoheptulosonate-7-phosphate synthetase (aroF) and feedback-resistant chorismate mutase/prephenate dehydratase (pheA ), led to increased productivity and decreased demand for aeration by E. coli CICC10245. Shake-flask studies showed that vgb-expressing strains displayed higher rates of oxygen uptake, and l-phenylalanine production under standard aeration conditions was increased. In the aerobic fermentation process, cell growth, l-phenylalanine production, and glucose consumption by the recombinant E. coli strain PAPV, which harbored aroF, pheA , and tac-vgb genes, were increased compared to that in the strain harboring only aroF and pheA (E. coli strain PAP), especially under oxygen-limited conditions. The vgb-expressing strain PAPV produced 21.9% more biomass and 16.6% more l-phenylalanine, while consuming only approximately 5% more glucose after 48 H of fermentation. This study demonstrates a method to enhance the l-phenylalanine production by E. coli using less intensive and thus more economical aeration conditions.

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Enhanced l-phenylalanine biosynthesis by co-expression of pheAfbr and aroFwt

Cited by 70 publications

References 31 publications

Enhanced l-phenylalanine production by recombinant Escherichia coli BR-42 (pAP-B03) resistant to bacteriophage BP-1 via a two-stage feeding approach

Enhanced l-phenylalanine production by recombinant Escherichia coli BR-42 (pAP-B03) resistant to bacteriophage BP-1 via a two-stage feeding approach

The influence of hydroxypropyl-β-cyclodextrin on the enantioselective hydrolysis of 2-amino phenylpropionitrile catalyzed by recombinant nitrilase

Enhancement of l‐phenylalanine production in Escherichia coli by heterologous expression of Vitreoscilla hemoglobin

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