Elevated intracellular acetyl-CoA availability by acs2 overexpression and mls1 deletion combined with metK1 introduction enhanced SAM accumulation in Saccharomyces cerevisiae

Chen, Hailong; Yang, Yang; Wang, Zhilai; Dou, Jun; Wang, Hui; Zhou, Changlin

doi:10.1016/j.bej.2015.11.016

Cited by 17 publications

(29 citation statements)

References 28 publications

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“…In previous works, several microorganisms were screened, and genetic strains were used to increase the production of SAM in fermentation processes [1]. In these methods, typically, L-methionine (L-Met) and ATP were supplied in the substrate, and SAM is synthesized by cellular S-adenosylmethionine synthetase [8,10,11]. The volumetric productivity of SAM was greatly improved after process optimization [1,2].…”

Section: Introductionmentioning

confidence: 99%

S-Adenosyl-l-methionine production by Saccharomyces cerevisiae SAM 0801 using dl-methionine mixture: From laboratory to pilot scale

Ren

Cai

et al. 2017

Process Biochemistry

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This study sought to develop a cost-effective biological catalysis process for S-adenosyl-L-methionine (SAM) production. During the process, a mixture of D-methionine (D-Met) and L-methionine (L-Met), namely DL-Met, was used as substrate to replace the conventional but expensive pure L-Met. The concentration of DL-Met in substrate was optimized. When 80 g/L of DL-Met was added in a 5 L-scale bioreactor, 13.74 g/L of SAM was produced with an L-Met conversion rate of 32.15%. The fermentation process was then scaled up to meet the requirements of realistic industrial SAM production. In a 300 L-scale fermentation process, 10.45 g/L of SAM was achieved, with an L-Met conversion rate of 32.61%. Moreover, the proportion of D-Met remaining in broth increased from 50% at the beginning to 76.89% at the end of fermentation. The fermentation process is generally appropriate for commercial SAM production.

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Section: Introductionmentioning

confidence: 99%

S-Adenosyl-l-methionine production by Saccharomyces cerevisiae SAM 0801 using dl-methionine mixture: From laboratory to pilot scale

Ren

Cai

et al. 2017

Process Biochemistry

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“…Currently, SAM is prepared industrially by high cell-density cultures of Saccharomyces cerevisiae or Pichia pastoris , which highly express MAT in the presence of exogenous l -methionine [ 9 , 10 , 11 , 12 ]. Genetic modification of yeast cells and the optimization of fermentation conditions are the two primary approaches to improving the production of SAM [ 13 , 14 , 15 , 16 , 17 , 18 ]. Zhao et al integrated endogenous MAT into an industrial strain to improve SAM accumulation.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, the addition of l -methionine in the medium is required for the enhancement of both SAM production as well as cell growth. Chen et al showed that the elevation of intracellular acetyl-CoA levels improved the intracellular methionine availability, which enhanced SAM accumulation (6.06 g/L) in yeast [ 13 ]. However, there are some obvious drawbacks to producing SAM via yeast fermentation, including a long fermentation period (48–120 h), high energy consumption, low SAM content and yield (~10 g/L medium), and a low conversion rate of l -methionine.…”

Section: Introductionmentioning

confidence: 99%

“…However, there are some obvious drawbacks to producing SAM via yeast fermentation, including a long fermentation period (48–120 h), high energy consumption, low SAM content and yield (~10 g/L medium), and a low conversion rate of l -methionine. Furthermore, the extraction and purification of SAM from yeast cells is complicated and environmentally unfriendly, since the yeast cells must be disrupted by first using a large amount of perchloric acid and then separating the SAM from the many other cell components [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

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Biosynthesis of S-Adenosylmethionine by Magnetically Immobilized Escherichia coli Cells Highly Expressing a Methionine Adenosyltransferase Variant

2017

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S-Adenosylmethionine (SAM) is a natural metabolite having important uses in the treatment of various diseases. To develop a simple and effective way to produce SAM, immobilized Escherichia coli cells highly expressing an engineered variant of methionine adenosyltransferase (MAT) were employed to synthesize SAM. The recombinant I303V MAT variant was successfully produced at approximately 900 mg/L in a 10-L bioreactor and exhibited significantly less product inhibition and had a four-fold higher specific activity (14.2 U/mg) than the wild-type MAT (3.6 U/mg). To reduce the mass transfer resistance, the free whole-cells were permeabilized and immobilized using gellan gum gel as support in the presence of 100 mg/L Fe3O4 nanoparticles, and the highest activity (4152.4 U/L support) was obtained, with 78.2% of the activity recovery. The immobilized cells were more stable than the free cells under non-reactive conditions, with a half-life of 9.1 h at 50 °C. Furthermore, the magnetically immobilized cells were employed to produce SAM at a 40-mM scale. The residual activity of the immobilized cells was 67% of its initial activity after 10 reuses, and the conversion rate of ATP was ≥95% in all 10 batches. These results indicated that magnetically immobilized cells should be a promising biocatalyst for the biosynthesis of SAM.

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“…Most studies have been focused on the improvement of SAM production by altering Saccharomyces cerevisiae or Pichia pastoris strains by a genetic engineering approach or by optimization of culture conditions [9][10][11][12][13][14][15][16]. However, the production of SAM by yeast fermentation has some obvious drawbacks, such as a long fermentation period, low SAM content and yield, and low conversion rate of the substrate.…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Synthesis of S-Adenosylmethionine Using Immobilized Methionine Adenosyltransferase Variants on the 50-mM Scale

et al. 2017

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S-adenosylmethionine (SAM), an important metabolite in all living organisms, has been widely used to treat various diseases. To develop a simple and efficient method to produce SAM, an engineered variant of the methionine adenosyltransferase (MAT) from Escherichia coli was investigated for its potential use in the enzymatic synthesis of SAM due to its significantly decreased product inhibition. The recombinant I303V MAT variant was successfully produced at a high level (~800 mg/L) with approximately four-fold higher specific activity than the wild-type MAT. The recombinant I303V MAT was covalently immobilized onto the amino resin and epoxy resin in order to obtain a robust biocatalyst to be used in industrial bioreactors. The immobilized preparation using amino resin exhibited the highest activity coupling yield (~84%), compared with approximately 3% for epoxy resin. The immobilized enzyme was more stable than the soluble enzyme under the reactive conditions, with a half-life of 229.5 h at 37 • C. The Km ATP value (0.18 mM) of the immobilized enzyme was ca. two-fold lower than that of the soluble enzyme. Furthermore, the immobilized enzyme showed high operational stability during 10 consecutive 8 h batches, with the substrate adenosine triphosphate (ATP) conversion rate above 95% on the 50-mM scale.

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Elevated intracellular acetyl-CoA availability by acs2 overexpression and mls1 deletion combined with metK1 introduction enhanced SAM accumulation in Saccharomyces cerevisiae

Cited by 17 publications

References 28 publications

S-Adenosyl-l-methionine production by Saccharomyces cerevisiae SAM 0801 using dl-methionine mixture: From laboratory to pilot scale

S-Adenosyl-l-methionine production by Saccharomyces cerevisiae SAM 0801 using dl-methionine mixture: From laboratory to pilot scale

Biosynthesis of S-Adenosylmethionine by Magnetically Immobilized Escherichia coli Cells Highly Expressing a Methionine Adenosyltransferase Variant

Enzymatic Synthesis of S-Adenosylmethionine Using Immobilized Methionine Adenosyltransferase Variants on the 50-mM Scale

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