Characterization of a vanillic acid non-oxidative decarboxylation gene cluster from Streptomyces sp. D7 The GenBank accession number for the sequence reported in this paper is AF134589.

Abstract

The genetics of non-oxidative decarboxylation of aromatic acids are poorly understood in both prokaryotes and eukaryotes. Although such reactions have been observed in numerous micro-organisms acting on a variety of substrates, the genes encoding enzymes responsible for these processes have not, to our knowledge, been reported in the literature. Here, the isolation of a streptomycete from soil (Streptomyces sp. D7) which efficiently converts 4-hydroxy-3-methoxybenzoic acid (vanillic acid) to 2-methoxyphenol (guaiacol) is described. Protein two-dimensional gel analysis revealed that several proteins were synthesized in response to vanillic acid. One of these was characterized by partial amino-terminal sequencing, leading to the cloning of a gene cluster from a genomic DNA lambda phage library, consisting of three ORFs, vdcB (602 bp), vdcC (1424 bp) and vdcD (239 bp). Protein sequence comparisons suggest that the product of vdcB (201 aa) is similar to phenylacrylate decarboxylase of yeast ; the putative products of vdcC (475 aa) and vdcD (80 aa) are similar to hypothetical proteins of unknown function from various micro-organisms, and are found in a similar cluster in Bacillus subtilis. Northern blot analysis revealed the synthesis of a 2 5 kb mRNA transcript in vanillic-acid-induced cells, suggesting that the cluster is under the control of a single inducible promoter. Expression of the entire vdc gene cluster in Streptomyces lividans 1326 as a heterologous host resulted in that strain acquiring the ability to decarboxylate vanillic acid to guaiacol non-oxidatively. Both Streptomyces sp. strain D7 and recombinant S. lividans 1326 expressing the vdc gene cluster do not, however, decarboxylate structurally similar aromatic acids, suggesting that the system is specific for vanillic acid. This catabolic system may be useful as a component for pathway engineering research focused towards the production of valuable chemicals from forestry and agricultural by-products.

Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica

“…Wiegel and coworkers identified the gene coding for the (reversible) 4-hydroxybenzoate decarboxylase (54.4 kDa) in Clostridium hydroxybenzoicum (30). This enzyme was similar to the three UbiD-like gene products of T. aromatica and to UbiD from E. coli (53.4 kDa) (17). Two other ORFs, orf2 and orf3, downstream of the gene for the 4-hydroxybenzoate decarboxylase were similar to the vdcDB genes of Streptomyces sp.…”

Metabolism of 2-hydroxy-1-naphthoic acid and naphthalene via gentisic acid by distinctly different sets of enzymes in Burkholderia sp. strain BC1

“…However, non-oxidative decarboxylation of various other aromatic acids such as vanillic acid (Chow et al, 1999;Lupa et al, 2008), 4-hydroxybenzoic acid (Lupa et al, 2008;Matsui et al, 2006), 3,4-dihydroxybenzoic acid (He &Wiegel, 1996) and 2,6-dihydroxybenzoic acid (Yoshida et al, 2004) has been studied extensively. Some of these non-oxidative decarboxylases were shown to catalyse reversible carboxylation reactions as well (Matsui et al, 2006;Yoshida et al, 2004).…”

Phenylphosphate Carboxylase: a New C-C Lyase Involved in Anaerobic Phenol Metabolism in Thauera aromatica

“…Also, a 54-kDa 4-hydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum (35) and subunits of various proven or putative vanillic acid decarboxylases from E. coli, Bacillus subtilis, and Streptomyces sp. show similarity to UbiD and UbiX (18,20,40), yet these enzymes are likely to function as decarboxylases, yielding phenolic compounds.…”