The Enzymatic Condensation of Pyruvate and Formaldehyde

Hift, Helen; Mahler, H. R.

doi:10.1016/s0021-9258(18)55548-4

Cited by 48 publications

(17 citation statements)

References 13 publications

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“…Wang at al. (2019) reported to use pyruvate as a HCHO acceptor catalyzed by 2-keto-4-hydroxybutyrate aldolase (KHB); − they further developed this reaction into a new 1,3-propanediol (PDO) biosynthesis pathway …”

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confidence: 99%

“…An acetaldehyde-dependent deoxyribose-5-phosphate aldolase from E. coli K-12 (DERA ECO ) turned out to well catalyze the condensation of HCHO and ACALD. 33 1.02 ± 0.03 mM (77.6 ± 2.28 mg/L) PDO were formed in a preliminary trial with 10 mM of HCHO and ACALD added after 2 h. To prove that the added HCHO and ACALD were condensed into the synthesis of PDO, we used 13 C-HCHO or 13 C-ACALD as cosubstrate for carrying out the biosynthesis. As shown in Figure 4 the ratio of the peaks around m/z 162.1 and m/z 163.1 are almost the same in the unlabeled standard sample and the 13 C-labled samples.…”

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confidence: 99%

“…33 1.02 ± 0.03 mM (77.6 ± 2.28 mg/L) PDO were formed in a preliminary trial with 10 mM of HCHO and ACALD added after 2 h. To prove that the added HCHO and ACALD were condensed into the synthesis of PDO, we used 13 C-HCHO or 13 C-ACALD as cosubstrate for carrying out the biosynthesis. As shown in Figure 4 the ratio of the peaks around m/z 162.1 and m/z 163.1 are almost the same in the unlabeled standard sample and the 13 C-labled samples. It confirmed that the synthesis of PDO totally depends on HCHO and ACALD added.…”

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confidence: 99%

“…39,40 PDO production with this new module was demonstrated in vitro using purified proteins, showing that HpADH1 can provide enough ACALD for the PDO formation (Figures S1C,D). We used 13 C-ethanol and HCHO as cosubstrates to verify the PDO production from such an ACALD supply module (Figure 6A). Using E. coli carrying DERA TMA and DhaT, ethanol and HCHO were successfully converted to PDO in a whole-cell bioconversion (Figure 6B).…”

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confidence: 99%

“…Indeed, several diols were formed in the bioconversion system using HCHO and ethanol as the substrates (Figure 7A). 13 C-ethanol was used as substrate to confirm the diol byproducts in batch bioconversion. As shown in Figure 7B−D, the GC-MS spectra of samples showed that the byproducts include 1,3-butanediol (1,3-BDO) (m/z 178.1), 3,5dihydroxypentanal (m/z 206.1), and 3,5-dihydroxyhexanal (m/ z 221.1).…”

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An Aldolase-Based New Pathway for Bioconversion of Formaldehyde and Ethanol into 1,3-Propanediol in Escherichia coli

et al. 2021

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Formaldehyde (HCHO) is a reactive one-carbon compound that is interesting for biosynthesis. The assimilation of HCHO depends on the catalysis of aldolase. Here, we present a novel synthetic pathway in E. coli to convert HCHO and ethanol into 1,3-propanediol (PDO) using a deoxyribose-5-phosphate aldolase (DERA). DERA condenses HCHO and acetaldehyde to form 3-hydroxypropionaldehyde, the direct precursor of PDO formation. This new pathway opens up the possibility to synthesize an appealing C3 compound from a C1 compound and a C2 compound without carbon loss in contrast to all the other known PDO synthetic pathways where typically 30–50% of the carbons are lost as CO2 and other byproducts. The pathway is successfully demonstrated by elaborating three metabolic modules. First, DERA from Thermotoga maritima was found to be efficient for the aldol condensation and PDO production module. For the module of acetaldehyde supply from ethanol, an alcohol dehydrogenase from Hansenula polymorpha was selected. For the HCHO supply module, the control of HCHO concentration and its utilization were shown to be important for achieving the assimilation of HCHO in recombinant E. coli cells. By deleting the gene frmA for endogenous conversion of HCHO to formate and controlling HCHO at a level of about 0.6 mM, the concentration and yield of PDO were increased from initially 5.67 mM (0.43 g/L) and 0.057 mol/mol to 17.35 mM (1.32 g/L) and 0.096 mol/mol in bioconversion of ethanol and HCHO with resting E. coli cells. Further engineering of DERA and the HCHO supply module is necessary to realize the potential of this promising metabolic pathway.

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