Metabolic engineering of <i>Escherichia coli</i> for microbial production of L‐methionine

Huang, Jianfeng; Liu, Zhi‐Qiang; Jin, Liqun; Tang, Xiaoling; Shen, Zhongliang; Yin, Huanhuan; Zheng, Yu‐Guo

doi:10.1002/bit.26198

Cited by 67 publications

(55 citation statements)

References 28 publications

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“…DL-Met (an equal racemic mixture of the D-and L-isomers of Met) is commonly added to poultry feeds that are low in natural Met to satisfy the requirement of chickens for this amino acid. However, only L-isomer that had been commercially available by a fermentation approach recently (Huang et al 2017), is the biologically functional form of Met, as it can be used readily in intestinal cells and directly incorporated into protein synthesis (Fang et al 2010). It is indispensable that dietary D-Met is converted to L-Met in a process catalysed by D-Met oxidase that resides in liver, kidney and intestines prior to its utilisation (Fang et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Bioavailability ofl-methionine relative todl-methionine in Broiler chickens

Wang

et al. 2019

Italian Journal of Animal Science

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This study was aimed to estimate the relative bioavailability (RBA) of L-methionine (L-Met) to DLmethionine (DL-Met) in broiler chickens from 1 to 21 d of age. A total of 1664 one-day-old broiler chicks were randomly divided into 13 groups with 8 replicates per group. The basal diet contained 60% of the NRC requirement for Met. Birds in treatment groups were fed the basal diet supplemented with graded levels of DL-or L-Met, resulting in the dietary content of Met at 75%, 80%, 85%, 90%, 95% and 100% of the NRC requirement, respectively. Results showed that there were linear increases (p < .05) in average daily gain (ADG), average daily feed intake, feed efficiency (FE), eviscerated weight, breast muscle weight and leg muscle weight in response to the increasing supplementation of either DL-or L-Met, which also linearly reduced (p < .05) plasma uric acid and protein carbonyl contents. Compared with DL-Met, L-Met addition resulted in elevations (p < .05) in ADG and FE, along with leg muscle yield of birds. A lower (p < .05) content of urea nitrogen with a higher (p < .05) content of uric acid in plasma were observed in the birds supplemented with L-Met compared with DL-Met. Slope-ratio assay revealed that the RBA of L-Met to DL-Met for ADG and FE were 141.5% and 189.1%, respectively. And the respective values were calculated as 122.9% and 116.8% when based on eviscerated weight and breast muscle weight as the response criteria. However, the above RBA estimates except for FE did not differ from 100%. Collectively, supplemental L-Met had a higher bioavailability for FE of broilers relative to DL-Met during the first 3 weeks of life. HIGHLIGHTS L-methionine addition has an advantage over DL-methionine for supporting the growth performance of broiler chickens in early age. The relative bioavailability of L-Met to DL-Met for feed efficiency of broiler chickens during the first 21 days post-hatch was calculated as 189.1% that differs from 100%.

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Section: Introductionmentioning

confidence: 99%

Bioavailability ofl-methionine relative todl-methionine in Broiler chickens

Wang

et al. 2019

Italian Journal of Animal Science

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“…Description Sources pTrc99a AmpR, pBR322-origin, trc promoter, 4.2 kb [35,38] pCas Kan R , Cas9 nuclease expression plasmid, temperature-sensitive origin [39,40] pTrc ΔrhtAΔilvIH/Gap-ilvA*-Pbs-leuDH without any obvious reduction of cell growth when promoters of different strengths including Pbs [30] and Gap from E. coli BL21 (DE3) were employed to regulate the expression of ilvA* and leuDH. These results indicated that the disruption of ilvIH drived more carbon flux to l-ABA and proper adjustments of attenuating the expression of ilvA* by a relatively weak promoter and enhancing the expression of leuDH by a strong promoter were also beneficial for the biosynthesis of l-ABA in this strain.…”

Section: Plasmidsmentioning

confidence: 99%

“…TPM medium was used to monitor the production of l-ABA, during the growth of cells at 35 [21]. The medium was adjusted to pH 7.0 by KOH.…”

Section: Strains Media and Growth Conditionsmentioning

confidence: 99%

“…Ammonia or phosphate was automatically fed into the broth to keep the pH at 7.0. When the glucose concentration in the broth was less than 5 g/L, 100 mL of feed solution (500 g/L glucose, 14 g/L (NH 4 ) 2 SO 4 , 12.5 g/L KH 2 PO 4 , 3 g/L l-methionine and 4.4 g/L l-lysine) was injected into the broth to raise the residual glucose concentration to around 20 g/L [21,35].…”

Section: Fermentationmentioning

confidence: 99%

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Fermentative production of the unnatural amino acid l-2-aminobutyric acid based on metabolic engineering

Zhang

et al. 2019

Microb Cell Fact

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Background l -2-aminobutyric acid ( l -ABA) is an unnatural amino acid that is a key intermediate for the synthesis of several important pharmaceuticals. To make the biosynthesis of l -ABA environmental friendly and more suitable for the industrial-scale production. We expand the nature metabolic network of Escherichia coli using metabolic engineering approach for the production of l -ABA. Results In this study, Escherichia coli THR strain with a modified pathway for threonine-hyperproduction was engineered via deletion of the rhtA gene from the chromosome. To redirect carbon flux from 2-ketobutyrate (2-KB) to l -ABA, the ilvIH gene was deleted to block the l -isoleucine pathway. Furthermore, the ilvA gene from Escherichia coli W3110 and the leuDH gene from Thermoactinomyces intermedius were amplified and co-overexpressed. The promoter was altered to regulate the expression strength of ilvA* and leuDH . The final engineered strain E. coli THR Δ rhtA Δ ilvIH /Gap- ilvA *-Pbs- leuDH was able to produce 9.33 g/L of l -ABA with a yield of 0.19 g/L/h by fed-batch fermentation in a 5 L bioreactor. Conclusions This novel metabolically tailored strain offers a promising approach to fulfill industrial requirements for production of l -ABA. Electronic supplementary material The online version of this article (10.1186/s12934-019-1095-z) contains supplementary material, which is available to authorized users.

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“…A comparative proteomics study between BL21 and W3110 manifested that W3110 maintained growth and metabolism at lower oxygen levels, thus enabling foreign protein to be gradually expressed [23]. Therefore, W3110 strain has also been applied to produce various chemicals, including L-methionine [24][25], L-homoserine [26], and L-malate [27].…”

Section: Introductionmentioning

confidence: 99%

Engineered chromosome-based T7 RNA polymerase in Escherichia coli W3110 for orthogonal T7 promoter circuit as a cell factory

Ting¹,

Tan²,

Ng³

2020

Preprint

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Background Orthogonal T7 RNA polymerase (T7RNAP) and T7 promoter were powerful tools to mediate the protein expression. Moreover, Escherichia coli W3110 strain possesses more advantages than the B strain due to more heat shock proteins and higher tolerance to chemicals. Therefore, implementation of T7-based system in W3110 strain is a conceivable strategy to develop the cell factory. Results Three novel W3110 strains with chromosome-equipped T7RNAP (i.e W3110:IL5, W3110::L5 and W3110::pI) were engineered to demonstrate the feasibility on protein expression and chemical production. At first, the LacZ and T7RNAP with IPTG induction showed higher expression levels in W3110 derivatives than that in BL21(DE3). The plasmids with and without lacI/lacO repression were used to investigate the protein expression of super-fold green fluorescence protein (sfGFP), Cas9, carbonic anhydrase (CA) and lysine decarboxylase (CadA). All the proteins were expressed higher and enzymatic functions were better in W3110::L5 and W3110::pI. Moreover, the highest cadaverine production, lysine consumption and the yield were obtained in W3110::L5(+) strain with pET28a(+)-CadA which reached 32.2 g/L, 45 g/L and 91.7% at 24 h, while the W3110::pI(-) strain with pSU-T7-CadA achieved 36.9 g/L, 43.8 g/L and 103.4% at 12 h which is unnecessary of inducer. Conclusion Inducer and lacI/lacO regulators on chromosome and plasmid have been investigated in W3110 strains with T7RNAP. The newly engineered W3110::L5 and W3110:pI both possessed similar protein expression compared to commercial BL21(DE3). Furthermore, among all strains, W3110::pI displayed the greatest potential as cell factory in the future.

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Metabolic engineering of Escherichia coli for microbial production of L‐methionine

Cited by 67 publications

References 28 publications

Bioavailability ofl-methionine relative todl-methionine in Broiler chickens

Bioavailability ofl-methionine relative todl-methionine in Broiler chickens

Fermentative production of the unnatural amino acid l-2-aminobutyric acid based on metabolic engineering

Engineered chromosome-based T7 RNA polymerase in Escherichia coli W3110 for orthogonal T7 promoter circuit as a cell factory

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