Increasing L-homoserine production in Escherichia coli by engineering the central metabolic pathways

Liu, Min; Lou, Jiali; Gu, Jiali; Lyu, Xiaomei; Wang, Fengqing; Wei, Dongzhi

doi:10.1016/j.jbiotec.2020.03.010

Cited by 16 publications

(26 citation statements)

References 30 publications

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“…coli W3110 in a fed-batch fermentation, three different publications reported a ca. 2-fold lower l -homoserine concentration (39.54, 35.8, and 37.57 g L –1 ) in a fed-batch fermentation after 40 h with l -homoserine yield on a glucose of ca. 0.29–0.35 g g –1 .…”

Section: Results and Discussionmentioning

confidence: 98%

“…It was found that l -alanine is also biotransformed and that the dependence of the reaction rate on l -alanine concentration ( r 8 ) can be described by the Michaelis–Menten kinetics with substrate inhibition (eq , Table ). In the longer experiments, it was observed that l -homoserine was also transformed, probably to l -threonine and l -methionine . The reaction rate of l -homoserine transformation ( r 9 ) was described by the Michaelis–Menten kinetics (eq , Table and Section S3, SI).…”

Section: Experimental Partmentioning

confidence: 99%

“…The downside of this technology is the formation of an unwanted acetic acid as a byproduct during the fermentation in concentrations of ca. 5.2–6.9 g L –1 , , which shows a toxic effect toward E. coli at concentrations of 5 g L –1 and higher .…”

Section: Introductionmentioning

confidence: 98%

“…l -Homoserine, synthesized in this work, is a noncanonical amino acid, a precursor of essential amino acids such as l -threonine, l -methionine, and l -isoleucine. − It is also a valuable building block for the synthesis of 3-hydroxypropionaldehyde and 1,3-propanediol − and has potential applications as a fertilizer and feed additive. , Several examples in the literature − reported the metabolic engineering of microorganisms for the production of l -homoserine. Metabolic engineering implies the use of growing cells as catalysts, which basically means that the biocatalyst is produced simultaneously as the reaction is carried out .…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Cascade Synthesis of l-Homoserine Catalyzed by Lyophilized Whole Cells Containing Transaminase and Aldolase Activities: The Mathematical Modeling Approach

Katulić

Sudar

Hernández

et al. 2021

Ind. Eng. Chem. Res.

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Aldolase and transaminase coexpressed in Escherichia coli cells and lyophilized (i.e., lyophilized whole-cell biocatalyst (LWCB)) were used as biocatalysts for the one-pot cascade synthesis of L-homoserine with substrate cycling. The kinetic analysis of enzymes within lyophilized cells was performed to evaluate the behavior of the system. The best result among the performed fed-batch reactor experiments achieved was 640.8 mM (76.3 g L −1 ) of L-homoserine with a volume productivity of 2.6 g L −1 h −1 . This is comparable with the results of the same cascade synthesis using cell-free extracts (CFEs) and significantly better than the reports in the literature applying fermentation technology. The approach applied here can serve as guidance for the design of microbial cells with an optimal ratio of expressed enzymes that act as biocatalysts in the cascade, resulting in lower biocatalyst cost, no need for the addition of expensive coenzymes, and enhanced enzyme stability as compared with cell-free extracts.

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Section: Results and Discussionmentioning

confidence: 98%

Section: Experimental Partmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Cascade Synthesis of l-Homoserine Catalyzed by Lyophilized Whole Cells Containing Transaminase and Aldolase Activities: The Mathematical Modeling Approach

Katulić

Sudar

Hernández

et al. 2021

Ind. Eng. Chem. Res.

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show abstract

“…First, the regulation of both glyoxylate shunt and TCA cycle in FAs is different from in glucose. The metabolic pro les of glyoxylate shunt and TCA cycle under FAsutilization condition should be systematically investigated 20,21,22 . To establish an e cient FAs feedstocks bioconversion process at large scale is another challenge.…”

Section: Introductionmentioning

confidence: 99%

Enhanced glyoxylate shunt for efficient production of C4 derivatives using fatty acids feedstocks

Miao

Liu

wang

et al. 2021

Preprint

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Biosynthesis of TCA cycle-derived C4 chemicals through glyoxylate shunt is an attractive metabolic route because it can be drived by TCA-glyoxylate cycle force under aerobic conditions. However, yield of this route is low with at least 1/3 carbon loss from glucose. FAs could sufficiently provide acetyl-CoA by β-oxidation without carbon loss and directly enter the TCA-glyoxylate cycle, which is acknowledged as a promising alternative feedstock. Here β-alanine was selected as the target TCA cycle-derived chemical, of which the theoretical yield is 1.391 g/g FAs, much higher than that of glucose(0.49 g/g). By adopting multi-metabolic engineering strategies and relieving the active oxygen damage caused by FAs utilization, β-alanine production reached 78.05 g/L with a yield of 1.2 g/g, about 86% of theoretical yield. Our study establish a promising bioproduction route of β-alanine from waste FAs (such as gutter oil, palm fatty acid distillate etc.), and more importantly, provide an efficient platform for TCA cycle-derived C4 chemicals biosynthesis.

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Engineering O‐Succinyl‐L‐Homoserine Mercaptotransferase for Efficient L‐Methionine Biosynthesis by Fermentation‐Enzymatic Coupling Route

Tang

Liu

et al. 2023

Adv Synth Catal

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L-Methionine is the unique sulfur-containing amino acid essential for humans and animals, which is widely used in pharmaceutical, food and feed industries. Its green and efficient production has attracted a great deal of attention. Fermentation-enzymatic coupling route is regarded to be promising for L-methionine biosynthesis, while O-succinyl-L-homoserine mercaptotransferase (MetZ) is the key biocatalyst. Exploring and engineering of MetZ with ideal catalytic properties is highly desired. Herein, the MetZ from Chromobacterium violaceum (CvMetZ) was screened and through in silico analyses, potential beneficial amino acid residues both at the access channel and around the oxygen anion holes were anchored for site-saturation mutagenesis. The positive mutants were obtained with improved activity for L-methionine biosynthesis using O-succinyl-Lhomoserine (OSH) and methyl mercaptan as substrate. The best mutant was further applied for L-methionine production and supply of methyl mercaptan was optimized in a fed-batch approach. The conversion of 100 g/L OSH reached 92% with L-methionine yield of 62.6 g/L, which was well coupled with the OSH fermentation level.

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Increasing L-homoserine production in Escherichia coli by engineering the central metabolic pathways

Cited by 16 publications

References 30 publications

Cascade Synthesis of l-Homoserine Catalyzed by Lyophilized Whole Cells Containing Transaminase and Aldolase Activities: The Mathematical Modeling Approach

Cascade Synthesis of l-Homoserine Catalyzed by Lyophilized Whole Cells Containing Transaminase and Aldolase Activities: The Mathematical Modeling Approach

Enhanced glyoxylate shunt for efficient production of C4 derivatives using fatty acids feedstocks

Engineering O‐Succinyl‐L‐Homoserine Mercaptotransferase for Efficient L‐Methionine Biosynthesis by Fermentation‐Enzymatic Coupling Route

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