Cloning and overexpression in Escherichia coli of the gene encoding dihydroxyacetone kinase isoenzyme I from Schizosaccharomyces pombe , and its application to dihydroxyacetone phosphate production

Tujibata, Y.; Liu, J. Q.

doi:10.1007/s002530051381

Cited by 27 publications

(16 citation statements)

References 13 publications

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“…13 Table 1 showing a good concordance with those published for cFMN. Finally, the structure elucidation of the minor compound was carried out.…”

Section: Nmr Proceduressupporting

confidence: 82%

“…[12] From a biocatalytic point of view, ATP-dependent DHAK's have been given considerable attention because their feasibility for the simple and efficient obtaining of DHAP. [13] We have reported a multi-enzyme system for one-pot C-C bond formation catalysed by DHAP-dependent aldolases, based in the use of the recombinant DHAK from C. freundii CECT 4626, for in situ DHAP formation. [14] This enzyme (GeneBank Accession Nº DQ473522) is a new isoform that differs in 22 aa from the enzyme of the DSM 30040 strain (see the Supporting Information).…”

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confidence: 99%

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From Kinase to Cyclase: An Unusual Example of Catalytic Promiscuity Modulated by Metal Switching

et al. 2009

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Enzyme promiscuity is a concept that in the last years is earning prominence in different fields of enzymology like biocatalysis, enzyme engineering or enzyme evolution. [1] Catalytic promiscuity is defined as the ability of an enzyme to catalyze more than one chemical transformation. [1b,c] Naturally occurring catalytic promiscuity provide the starting point for a Darwinian evolution of enzymes to new functions since this process must occur gradually, while maintaining organism fitness throughout. [2] Tawfik and co-workers [3] have provided experimental evidence for the plasticity and "evolvability" [4] of promiscuous functions. These authors propose a model by which a protein acquires a new function, without losing the original one, and gene duplication may follow the emergence of a new function, rather than initiate it. Besides the intriguing implications that this theory of divergent molecular evolution has for protein evolution, its application to promiscuous enzymes allows to design enzymes with new catalytic activities. [5] A special case of catalytic promiscuity is the shown by metalloenzymes, where the variety of metallic ions that can be incorporated in the active site increase the range of chemical transformations that can be catalyzed by the enzyme. Thus, there are several examples showing that protein modification via the covalent attachment of ligands that incorporate metal ions or by incorporation of the catalytic metal ion alone in a suitable site for coordination, is a strategy that allows to obtain enzymes with either modified or completely new catalytic activities. [6] In this communication we describe the promiscuous behaviour of the dihydroxyacetone (DHA) kinase from Citrobacter freundii strain CECT 4626. This ATP-dependent DHAK is able to catalyse, beside the transfer of the γ-phosphate of the ATP to DHA, the cyclization of the FAD to yield riboflavin 4',5'-cyclic phosphate (4',5'-cFMN) (Scheme 1). This catalytic promiscuity is modulated by the divalent cation that forms the complex with the phosphorylated substrate. Although DHAK's are widely distributed in the three biological kingdoms, only their role in the catabolism of glycerol and in methanol assimilation in microorganisms have been well characterized. [7] In the bacteria C. freundii strain DSM 30040 [8] the entire dha regulon has been cloned and characterized at molecular level. [9] The kinetic properties and mechanism of the corresponding DHAK have been described [10] and the X-ray structure of the full-length DHAK in complex with its substrates has been elucidated. [11] This kinase is the only one known with an all-α nucleotide-binding domain. [12] From a biocatalytic point of view, ATP-dependent DHAK's have been given considerable attention because their feasibility for the simple and efficient obtaining of DHAP. [13] We have reported a multi-enzyme system for one-pot C-C bond formation catalysed by DHAP-dependent aldolases, based in the use of the recombinant DHAK from C. freundii CECT 4626, for in situ DHAP formation. [14...

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“…13 Table 1 showing a good concordance with those published for cFMN. Finally, the structure elucidation of the minor compound was carried out.…”

Section: Nmr Proceduressupporting

confidence: 82%

mentioning

confidence: 99%

From Kinase to Cyclase: An Unusual Example of Catalytic Promiscuity Modulated by Metal Switching

et al. 2009

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“…However, to make the process more practical, energy (ATP) should be supplied in more economical forms and recycled. Although production of DR5P from glucose requires ATP, there are some ATP-providing or -regenerating systems (for example, ATP production from adenine by Corynebacterium ammoniagenes [12] or from adenosine by a methylotrophic yeast [17] and ATP regeneration by acetate kinase [9] or by baker's yeast through alcoholic fermentation [23]) which could be coupled with DR5P synthesis from glucose and acetaldehyde via G3P. Coupling of such energy generation systems is currently under investigation.…”

Section: Discussionmentioning

confidence: 99%

Construction of Deoxyriboaldolase-Overexpressing Escherichia coli and Its Application to 2-Deoxyribose 5-Phosphate Synthesis from Glucose and Acetaldehyde for 2′-Deoxyribonucleoside Production

Horinouchi

Ogawa

Sakai

et al. 2003

Appl Environ Microbiol

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The gene encoding a deoxyriboaldolase (DERA) was cloned from the chromosomal DNA of Klebsiella pneumoniae B-4-4. This gene contains an open reading frame consisting of 780 nucleotides encoding 259 amino acid residues. The predicted amino acid sequence exhibited 94.6% homology with the sequence of DERA from Escherichia coli. The DERA of K. pneumoniae was expressed in recombinant E. coli cells, and the specific activity of the enzyme in the cell extract was as high as 2.5 U/mg, which was threefold higher than the specific activity in the K. pneumoniae cell extract. One of the E. coli transformants, 10B5/pTS8, which had a defect in alkaline phosphatase activity, was a good catalyst for 2-deoxyribose 5-phosphate (DR5P) synthesis from glyceraldehyde 3-phosphate and acetaldehyde. The E. coli cells produced DR5P from glucose and acetaldehyde in the presence of ATP. Under the optimal conditions, 100 mM DR5P was produced from 900 mM glucose, 200 mM acetaldehyde, and 100 mM ATP by the E. coli cells. The DR5P produced was further transformed to 2-deoxyribonucleoside through coupling the enzymatic reactions of phosphopentomutase and nucleoside phosphorylase. These results indicated that production of 2-deoxyribonucleoside from glucose, acetaldehyde, and a nucleobase is possible with the addition of a suitable energy source, such as ATP.There will be a need for 2Ј-deoxyribonucleoside in the near future due to increasing demand in new medical and biotechnology fields. 2Ј-Deoxyribonucleoside is a building block of promising antisense drugs for cancer therapy. For some recently developed antiviral agents, such as azidothymidine for treatment of human immunodeficiency virus infections, 2Ј-deoxyribonucleoside is a synthesis intermediate. 2Ј-Deoxyribonucleoside is also a precursor of an indispensable material used for widespread PCR applications, 2Ј-deoxyribonucleoside triphosphate. The current 2Ј-deoxyribonucleoside sources include hydrolyzed herring and salmon sperm DNA, which are not suitable sources for sudden high demand. The difficulty in chemical synthesis of 2Ј-deoxyribonucleoside lies in the generation of 2-deoxyribosyl groups. The chemical synthesis of 2-deoxyribosyl groups and the subsequent synthesis of 2Ј-deoxyribonucleoside involve complex protection and deprotection steps (1,8,10,15). It is likely that introduction of biochemical reactions with high selectivity will solve this problem.In general metabolism, there are two reactions involving 2-deoxyriobosyl groups. One of these reactions is reduction of ribonucleotide to 2Ј-deoxyribonuleotide during biosynthesis, and the other is cleavage of 2-deoxyribose 5-phosphate (DR5P) to produce glyceraldehyde 3-phosphate (G3P) and acetaldehyde during degradation of 2Ј-deoxyribonucleoside. The former reaction is regulated in a complicated fashion and is hard to handle for practical purposes (3, 4). We focused on the latter reaction for generation of a 2-deoxyribose frame and found that the reverse reactions of 2Ј-deoxyribonucleoside degradation are promising reactions for synth...

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“…The Dha-specific kinases have been characterized in bacteria [10,41] and in methylotrophic yeast (for a review see [42][43][44]). They have been purified from yeasts [16,17,42,43,[45][46][47][48][49], algae [50], tomato [51], rat brain [31], porcine kidney [52] and from bacteria, viz. C. freundii [10,41], K. pneumoniae [13] and E. coli [53].…”

Section: Occurrence and Biochemistry Of Dha Kinasesmentioning

confidence: 99%

Small substrate, big surprise: fold, function and phylogeny of dihydroxyacetone kinases

Erni

Siebold

Christen

et al. 2006

Cell. Mol. Life Sci.

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Dihydroxyacetone (Dha) kinases are a family of sequence-conserved enzymes which utilize either ATP (in animals, plants and eubacteria) or phosphoenolpyruvate (PEP, in eubacteria) as their source of high-energy phosphate. The kinases consist of two domains/subunits: DhaK, which binds Dha covalently in hemiaminal linkage to the Nepsilon2 of a histidine, and DhaL, an eight-helix barrel that contains the nucleotide-binding site. The PEP-dependent kinases comprise a third subunit, DhaM, which rephosphorylates in situ the firmly bound ADP cofactor. DhaM serves as the shuttle for the transfer of phosphate from the bacterial PEP: carbohydrate phosphotransferase system (PTS) to the Dha kinase. The DhaL and DhaK subunits of the PEP-dependent Escherichia coli kinase act as coactivator and corepressor of DhaR, a transcription factor from the AAA(+) family of enhancerbinding proteins. In Gram-positive bacteria genes for homologs of DhaK and DhaL occur in operons for putative transcription factors of the TetR and DeoR families. Proteins with the Dha kinase fold can be classified into three families according to phylogeny and function: Dha kinases, DhaK and DhaL homologs (paralogs) associated with putative transcription regulators of the TetR and DeoR families, and proteins with a circularly permuted domain order that belong to the DegV family.

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Cloning and overexpression in Escherichia coli of the gene encoding dihydroxyacetone kinase isoenzyme I from Schizosaccharomyces pombe , and its application to dihydroxyacetone phosphate production

Cited by 27 publications

References 13 publications

From Kinase to Cyclase: An Unusual Example of Catalytic Promiscuity Modulated by Metal Switching

From Kinase to Cyclase: An Unusual Example of Catalytic Promiscuity Modulated by Metal Switching

Construction of Deoxyriboaldolase-Overexpressing Escherichia coli and Its Application to 2-Deoxyribose 5-Phosphate Synthesis from Glucose and Acetaldehyde for 2′-Deoxyribonucleoside Production

Small substrate, big surprise: fold, function and phylogeny of dihydroxyacetone kinases

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