Efficient Production of 2-Deoxyribose 5-Phosphate from Glucose and Acetaldehyde by Coupling of the Alcoholic Fermentation System of Baker’s Yeast and Deoxyriboaldolase-Expressing<i>Escherichia coli</i>

Horinouchi, Nobuyuki; Ogawa, Jun; Kawano, Takako; Sakai, Takafumi; Saito, Kyota; Matsumoto, Seiichiro; Sasaki, Makiko; Mikami, Yoichi; Shimizu, Sakayu

doi:10.1271/bbb.50648

Cited by 20 publications

(10 citation statements)

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“…With the aim of evaluating another glycolytic metabolite, a simple methodology previously reported for FDP production was employed [22]. The reaction mixture containing FDP from baker's yeast was directly used as the starting material for the …”

Section: Resultsmentioning

confidence: 99%

“…The reaction mixture comprising 13% (w/v) biocatalyst, 50% (v/v) fructose FDP solution obtained according to Horinouchi et al [22], 400 mM acetaldehyde, and 1% (v/v) xylene (final volume 1 mL) was stirred at 200 rpm and 28°C for 3 h. Samples were centrifuged at 11,000 rpm for 3 min, and the supernatants were analyzed by TLC.…”

Section: Dr5p Preparation From Fructose 16-diphosphate (Mr3)mentioning

confidence: 99%

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Selection of a New Whole Cell Biocatalyst for the Synthesis of 2-Deoxyribose 5-Phosphate

Valino

Palazzolo

Iribarren

et al. 2011

Appl Biochem Biotechnol

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2-deoxyribose 5-phosphate (DR5P) is a key intermediate in the biocatalyzed preparation of deoxyribonucleosides. Therefore, DR5P production by means of simpler, cleaner, and economic pathways becomes highly interesting. One strategy involves the use of bacterial whole cells containing DR5P aldolase as biocatalyst for the aldol addition between acetaldehyde and D: -glyceraldehyde 3-phosphate or glycolytic intermediates that in situ generate the acceptor substrate. In this work, diverse microorganisms capable of synthesizing DR5P were selected by screening several bacteria genera. In particular, Erwinia carotovora ATCC 33260 was identified as a new biocatalyst that afforded 14.1-mM DR5P starting from a cheap raw material like glucose.

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Section: Resultsmentioning

confidence: 99%

Section: Dr5p Preparation From Fructose 16-diphosphate (Mr3)mentioning

confidence: 99%

Selection of a New Whole Cell Biocatalyst for the Synthesis of 2-Deoxyribose 5-Phosphate

Valino

Palazzolo

Iribarren

et al. 2011

Appl Biochem Biotechnol

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“…1) The excellent ATP regeneration system. 2) Baker’s yeast cell treated by organic solvent (toluene or acetone) temporarily accumulate fructose 1,6-diphosphate (FDP) out of a cell under existence of glucose and phosphate [4-6]. In order to construct a microbial platform for an energy-requiring bioprocess, we established an efficient enzymatic process in which 2′-deoxyribonucleosides (dNSs) are synthesized from glucose, acetaldehyde, and a nucleobase via a C−C coupling reaction requiring a high-energy substrate provided by the glycolytic pathway as an energy-generating system (Figure 1c).…”

Section: Introductionmentioning

confidence: 99%

“…c) An example of energy-requiring bioprocesses: Microbial production of dNS from glucose, acetaldehyde, and a nucleobase. In this process, FDP generated from glucose by baker’s yeast [4,9] serves as a substrate for fructose 1,6-diphosphate aldolase (FDP ALD) reaction in E. coli (DERA-PPMase−co-expressing E. coli ), and then the generated triose phosphates (DHAP and G3P) are converted to dNS through reactions catalyzed by triose phosphate isomerase (TPI) [10] and the enzymes involved in dNS metabolism (DERA-PPMase-NPase) [9,11,12]. This process is classified into type I in Figure 1a.…”

Section: Introductionmentioning

confidence: 99%

Construction of microbial platform for an energy-requiring bioprocess: practical 2′-deoxyribonucleoside production involving a C−C coupling reaction with high energy substrates

et al. 2012

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BackgroundReproduction and sustainability are important for future society, and bioprocesses are one technology that can be used to realize these concepts. However, there is still limited variation in bioprocesses and there are several challenges, especially in the operation of energy-requiring bioprocesses. As an example of a microbial platform for an energy-requiring bioprocess, we established a process that efficiently and enzymatically synthesizes 2′-deoxyribonucleoside from glucose, acetaldehyde, and a nucleobase. This method consists of the coupling reactions of the reversible nucleoside degradation pathway and energy generation through the yeast glycolytic pathway.ResultsUsing E. coli that co-express deoxyriboaldolase and phosphopentomutase, a high amount of 2′-deoxyribonucleoside was produced with efficient energy transfer under phosphate-limiting reaction conditions. Keeping the nucleobase concentration low and the mixture at a low reaction temperature increased the yield of 2′-deoxyribonucleoside relative to the amount of added nucleobase, indicating that energy was efficiently generated from glucose via the yeast glycolytic pathway under these reaction conditions. Using a one-pot reaction in which small amounts of adenine, adenosine, and acetone-dried yeast were fed into the reaction, 75 mM of 2′-deoxyinosine, the deaminated product of 2′-deoxyadenosine, was produced from glucose (600 mM), acetaldehyde (250 mM), adenine (70 mM), and adenosine (20 mM) with a high yield relative to the total base moiety input (83%). Moreover, a variety of natural dNSs were further synthesized by introducing a base-exchange reaction into the process.ConclusionA critical common issue in energy-requiring bioprocess is fine control of phosphate concentration. We tried to resolve this problem, and provide the convenient recipe for establishment of energy-requiring bioprocesses. It is anticipated that the commercial demand for dNSs, which are primary metabolites that accumulate at very low levels in the metabolic pool, will grow. The development of an efficient production method for these compounds will have a great impact in both fields of applied microbiology and industry and will also serve as a good example of a microbial platform for energy-requiring bioprocesses.

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“…Because the reaction catalyzed by DERA is reversible, the decrease of G3P results in a shift in equilibrium toward the cleavage of DR5P. It is also known that DR5P is dephosphorylated at 37 • C or higher (Horinouchi et al, 2006b).…”

mentioning

confidence: 99%

Production of 2-deoxyribose 5-phosphate from fructose to demonstrate a potential of artificial bio-synthetic pathway using thermophilic enzymes

Honda

Maya

Ômasa

et al. 2010

Journal of Biotechnology

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Efficient Production of 2-Deoxyribose 5-Phosphate from Glucose and Acetaldehyde by Coupling of the Alcoholic Fermentation System of Baker’s Yeast and Deoxyriboaldolase-ExpressingEscherichia coli

Cited by 20 publications

References 20 publications

Selection of a New Whole Cell Biocatalyst for the Synthesis of 2-Deoxyribose 5-Phosphate

Selection of a New Whole Cell Biocatalyst for the Synthesis of 2-Deoxyribose 5-Phosphate

Construction of microbial platform for an energy-requiring bioprocess: practical 2′-deoxyribonucleoside production involving a C−C coupling reaction with high energy substrates

Production of 2-deoxyribose 5-phosphate from fructose to demonstrate a potential of artificial bio-synthetic pathway using thermophilic enzymes

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