4-<i>N</i>-Trimethylaminobutyraldehyde Dehydrogenase: Purification and Characterization of an Enzyme from<i>Pseudomonas</i>sp. 13CM

Hassan, Mohd Rohaizat; Okada, Masahiro; Ichiyanagi, Tsuyoshi; Mori, Nobuhiro

doi:10.1271/bbb.70514

Cited by 7 publications

(4 citation statements)

References 59 publications

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“…Like the case of ω‐aminoaldehydes, BBD1 exhibited the lower affinity to these N ‐trimethylaminoaldehydes but higher specific activities than BBD2 (Table 3). TMAB‐ald is the intermediate of carnitine synthesis in mammals and some microorganisms (Hassan et al 2008, Vaz and Wanders 2002). It was also reported that carnitine was present in barley leaves and stimulated the rate of Chl production in etiolated barley leaves exposed to light (Ariffin et al 1982, Thomas et al 1981).…”

Section: Resultsmentioning

confidence: 99%

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Enzymatic characterization of peroxisomal and cytosolic betaine aldehyde dehydrogenases in barley

Fujiwara

Hori

Ozaki

et al. 2008

Physiologia Plantarum

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Betaine aldehyde dehydrogenase (BADH; EC 1.2.1.8) is an important enzyme that catalyzes the last step in the synthesis of glycine betaine, a compatible solute accumulated by many plants under various abiotic stresses. In barley (Hordeum vulgare L.), we reported previously the existence of two BADH genes (BBD1 and BBD2) and their corresponding proteins, peroxisomal BADH (BBD1) and cytosolic BADH (BBD2). To investigate their enzymatic properties, we expressed them in Escherichia coli and purified both proteins. Enzymatic analysis indicated that the affinity of BBD2 for betaine aldehyde was reasonable as other plant BADHs, but BBD1 showed extremely low affinity for betaine aldehyde with apparent K(m) of 18.9 microM and 19.9 mM, respectively. In addition, V(max)/K(m) with betaine aldehyde of BBD2 was about 2000-fold higher than that of BBD1, suggesting that BBD2 plays a main role in glycine betaine synthesis in barley plants. However, BBD1 catalyzed the oxidation of omega-aminoaldehydes such as 4-aminobutyraldehyde and 3-aminopropionaldehyde as efficiently as BBD2. We also found that both BBDs oxidized 4-N-trimethylaminobutyraldehyde and 3-N-trimethylaminopropionaldehyde.

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Section: Resultsmentioning

confidence: 99%

“…Although TMAB‐ald dehydrogenase has been purified to homogeneity from Pseudomona s sp. 13CM and rat liver (Hassan et al 2008, Vaz et al 2000), it has never reported from plants. Our data imply the additional function of BBD1 and BBD2.…”

Section: Resultsmentioning

confidence: 99%

Enzymatic characterization of peroxisomal and cytosolic betaine aldehyde dehydrogenases in barley

Fujiwara

Hori

Ozaki

et al. 2008

Physiologia Plantarum

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“…In the third step, the NAD + -dependent TMABA dehydrogenase (TMABADH; EC 1.2.1.47) oxidizes TMABA to γ-butyrobetaine (γ-BB) as shown for Pseudomonas sp. 13CM (Hassan et al, 2008;Bari et al, 2013). Finally, stereoselective hydroxylation of γ-BB by γ-BB hydroxylase (γ-BBH; EC 1.14.11.1) yields L-carnitine.…”

Section: Introductionmentioning

confidence: 99%

L-Carnitine Production Through Biosensor-Guided Construction of the Neurospora crassa Biosynthesis Pathway in Escherichia coli

Kugler

Trumm

Frese

et al. 2021

Front. Bioeng. Biotechnol.

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L-Carnitine is a bioactive compound derived from L-lysine and S-adenosyl-L-methionine, which is closely associated with the transport of long-chain fatty acids in the intermediary metabolism of eukaryotes and sought after in the pharmaceutical, food, and feed industries. The L-carnitine biosynthesis pathway has not been observed in prokaryotes, and the use of eukaryotic microorganisms as natural L-carnitine producers lacks economic viability due to complex cultivation and low titers. While biotransformation processes based on petrochemical achiral precursors have been described for bacterial hosts, fermentative de novo synthesis has not been established although it holds the potential for a sustainable and economical one-pot process using renewable feedstocks. This study describes the metabolic engineering of Escherichia coli for L-carnitine production. L-carnitine biosynthesis enzymes from the fungus Neurospora crassa that were functionally active in E. coli were identified and applied individually or in cascades to assemble and optimize a four-step L-carnitine biosynthesis pathway in this host. Pathway performance was monitored by a transcription factor-based L-carnitine biosensor. The engineered E. coli strain produced L-carnitine from supplemented L-Nε-trimethyllysine in a whole cell biotransformation, resulting in 15.9 μM carnitine found in the supernatant. Notably, this strain also produced 1.7 μM L-carnitine de novo from glycerol and ammonium as carbon and nitrogen sources through endogenous Nε-trimethyllysine. This work provides a proof of concept for the de novoL-carnitine production in E. coli, which does not depend on petrochemical synthesis of achiral precursors, but makes use of renewable feedstocks instead. To the best of our knowledge, this is the first description of L-carnitine de novo synthesis using an engineered bacterium.

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“…that NAD + -dependent TMA-butanol dehydrogenase (TMA-butanol DH) and 4-trimethylaminobutyraldehyde dehydrogenase (TMA-butyraldehyde DH) are involved in the TMAbutanol degradation pathway. [14][15][16] Those two enzymes were characterized, and those genes were cloned. 17) However, eukaryotic microbes able to grow on homocholine and TMA-butanol have not been found.…”

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confidence: 99%

Purification and characterization of 4-N-trimethylamino-1-butanol dehydrogenase from Fusarium merismoides var. acetilereum

Fujimitsu

Taniyama

Tajima

et al. 2016

Bioscience, Biotechnology, and Biochemistry

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From investigation of 60 filamentous fungi, we identified Fusarium merismoides var. acetilereum, which uses 4-N-trimethylamino-1-butanol (TMA-butanol) as the sole source of carbon and nitrogen. The fungus produced NAD(+)-dependent TMA-butanol dehydrogenase (DH) when it was cultivated in medium containing TMA-butanol. The enzyme showed molecular mass of 40 kDa by SDS-PAGE and 160 kDa by gel filtration, suggesting that it is a homotetramer. TMA-butanol DH is stable at pH 7.5-9.0. It exhibits moderate stability with respect to temperature (up to 30 °C). Additionally, it has optimum activity at 45 °C and at pH 9.5. The enzyme has broad specificity to various alkyl alcohols and amino alkyl alcohols, and the carbon chains of which are longer than butanol. Moreover, the activity is strongly inhibited by oxidizing agents, carbonyl and thiol modulators, and chelating agents. This report is the first study examining TMA-butanol DH from eukaryotic microbes.

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4-N-Trimethylaminobutyraldehyde Dehydrogenase: Purification and Characterization of an Enzyme fromPseudomonassp. 13CM

Cited by 7 publications

References 59 publications

Enzymatic characterization of peroxisomal and cytosolic betaine aldehyde dehydrogenases in barley

Enzymatic characterization of peroxisomal and cytosolic betaine aldehyde dehydrogenases in barley

L-Carnitine Production Through Biosensor-Guided Construction of the Neurospora crassa Biosynthesis Pathway in Escherichia coli

Purification and characterization of 4-N-trimethylamino-1-butanol dehydrogenase from Fusarium merismoides var. acetilereum

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