Growth of Mycobacteria on Carbon Monoxide and Methanol

Abstract

Carboxydobacteria are a group of bacteria which are able to grow chemolithotrophically on carbon monoxide (CO) as the sole carbon and energy source under aerobic conditions (22,30 Microbiol. 1965Microbiol. , abstr. P108, 1965, and Actinoplanes, Microbispora, and Mycobacterium (4), have also been described.The facultatively chemolithotrophic bacterium Mycobacterium sp. strain JC1 (originally Acinetobacter sp. strain JC1 DSM 3803; reclassified by Song et al. [41]), is capable of growing aerobically not only on CO but also on methanol as a sole source of carbon and energy (8,39). This means that the bacterium is able to employ three distinct types of nutrition, chemoheterotrophy, chemolithotrophy, and methylotrophy, depending on substrate availability.Combined with these results, the facts that many mycobacterial species including Mycobacterium tuberculosis (10; GenBank accession no. AL123456), Mycobacterium avium (NCBI reference sequence [RefSeq] NC-002943), Mycobacterium bovis (NCBI RefSeq NC-002945), Mycobacterium leprae (9; GenBank accession no. AL450380), and Mycobacterium smegmatis (NCBI RefSeq NC-002974) have genes encoding amino acid sequences similar to those of Mycobacterium sp. strain JC1 CO dehydrogenase (CO-DH) (T. Song and Y. M. Kim, unpublished data), that Mycobacterium phlei is able to oxidize CO (4), and that Mycobacterium cuneatum (40), Mycobaterium gastri (18), and Mycobacterium ID-Y (36) are capable of growing on methanol raise the possibility that all known mycobacteria have an intrinsic ability to grow on CO and/or methanol as the sole carbon and energy source.In order to address this question, we examined several wellknown mycobacteria for the ability to grow on CO and/or methanol, and we found that all the mycobacteria tested grew well on each of these substrates as the sole source of carbon and energy, except that M. tuberculosis did not grow on methanol. We also present several enzymological backgrounds for the growth of the mycobacteria on CO and methanol. MATERIALS AND METHODSStrains and cultivation conditions. Mycobacterium sp. strain JC1 (DSM 3803) (3, 41), Mycobacterium flavescens (ATCC 14474), M. gastri (ATCC 15754), Mycobacterium neoaurum (ATCC 25795), Mycobacterium parafortuitum (ATCC 19686), Mycobacterium peregrinum (ATCC 14467), M. phlei (ATCC 11758), M. smegmatis mc 2 (ATCC 700084), M. tuberculosis H37Ra (ATCC 35835), and Mycobacterium vaccae (ATCC 15483) were used throughout this study. Cells were cultivated at 37°C under CO chemolithoautotrophy with a gas mixture of 30% CO-70% air in either standard mineral base (SMB) medium (SMB-CO) (21) or 0.47% (wt/vol) Middlebrook 7H9 medium (7H9-CO; Becton Dickinson, Cockeysville, Md.). For methylotrophic growth, cells were grown at 37°C in SMB medium supplemented with 1% (vol/vol) methanol (SMB-MeOH). For the methanol assimilation enzyme assay, Methylobacterium extorquens AM1 (NCIB 9133) and Methylobacillus sp. strain SK1 (DSM 8269) grown at 30°C in SMBMeOH were used as controls. Growth was measured with a spectrophotometer by determi...

Cloning and expression analysis of the duplicated genes for carbon monoxide dehydrogenase of Mycobacterium sp. strain JC1 DSM 3803

“…JC1 has no epitopes identical to those of the non-mycobacterial CO-DHs . Several mycobacterial species have been shown to consume CO (King, 2003;Park et al, 2003) and to have nucleotide sequences corresponding to a part of the large subunit of CO-DH (King, 2003). Phylogenetic analyses with a part of the deduced amino acid sequences of the tentative large subunit genes of CO-DHs in other mycobacteria have also suggested that mycobacterial CO-DHs form a subclade distinct from other CO-DHs in other bacteria (King, 2003).…”

“…CO-DH from the bacterium is similar in molecular mass, subunit structure, cofactor composition and other properties to the enzymes of Gram-negative carboxydobacteria (Gram-negative CO-DH), although not immunologically related to them . We have also shown that several mycobacterial species are capable of carboxydotrophic growth and possess CO-DH enzymes that share common antigenic epitopes (Park et al, 2003). In addition, we have recently demonstrated that the CO-DHs from several mycobacteria, including Mycobacterium sp.…”

Genes involved in the methyl tert-butyl ether (MTBE) metabolic pathway of Mycobacterium austroafricanum IFP 2012

“…Seven of these proteins were characterized: (i) proteins of 64, 55 and 27 kDa encoded by the mpdB, mpdC and orf1 genes, respectively, described in this study; (ii) proteins of 67, 63 and 11 kDa corresponding to DnaK and GroEL/ES molecular chaperones, respectively; and (iii) a protein of 50 kDa, which is the methanol : NDMA oxidoreductase detected in Mycobacterium gastri (Bystrykh et al, 1993) and several other mycobacteria (Park et al, 2003). This latter protein was detected in crude extracts of M. austroafricanum IFP 2012 after growth on HIBA and 2-propanol (François, 2002), suggesting its involvement in assimilation of 2-propanol, a putative intermediate of the MTBE metabolic pathway.…”

Identification and functional characterization of a gene for the methanol : N,N'-dimethyl-4-nitrosoaniline oxidoreductase from Mycobacterium sp. strain JC1 (DSM 3803)

“…strain JC1 grown with methanol did not exhibit activities of the classical PQQ-containing MDH in Gram-negative bacteria, the NAD-dependent PQQ-containing MDH and cytochrome c-dependent MDH of A. methanolica, the NADdependent MDH of methylotrophic Bacillus, or the NADdependent glutathionine-requiring MDH and methanol oxidase of methylotrophic yeasts (Anthony & Zatman, 1967;Arfman et al, 1989;Duine et al, 1984;Kato et al, 1975;Mehta, 1975;Tani et al, 1972). Instead, the bacterium exhibited MDO activity during growth on methanol (Park et al, 2003), suggesting that this bacterium may use MDO as a key enzyme for the oxidation of methanol. However, it has not been identified whether MDO is the enzyme necessary for Mycobacterium sp.…”

“…strain JC1 can grow on methanol as the sole source of carbon and energy (Cho et al, 1985;Ro et al, 1997b;Song et al, 2002). In previous experiments (Park et al, 2003;Ro et al, 1997a), Mycobacterium sp. strain JC1 grown with methanol did not exhibit activities of the classical PQQ-containing MDH in Gram-negative bacteria, the NAD-dependent PQQ-containing MDH and cytochrome c-dependent MDH of A. methanolica, the NADdependent MDH of methylotrophic Bacillus, or the NADdependent glutathionine-requiring MDH and methanol oxidase of methylotrophic yeasts (Anthony & Zatman, 1967;Arfman et al, 1989;Duine et al, 1984;Kato et al, 1975;Mehta, 1975;Tani et al, 1972).…”