Gene Expression Analysis of Methylotrophic Oxidoreductases Involved in the Oligotrophic Growth of<i>Rhodococcus erythropolis</i>N9T-4

Yoshida, Nobuyuki; Hayasaki, Takuya; Takagi, Hiroshi

doi:10.1271/bbb.100700

Cited by 21 publications

(11 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In C. glutamicum, mycothiol was reported to contribute to resistance to alkylating agents, glyphosate, ethanol, antibiotics, heavy metals, and aromatic compounds (63). Our findings indicate that C. glutamicum, similar to R. erythropolis (47), can oxidize formaldehyde in an MSH-dependent manner via AdhE and in an NAD-dependent manner via Ald.…”

Section: Discussionmentioning

confidence: 65%

“…Mean values are shown. smegmatis (46), and Rhodococcus erythropolis (47). These proteins catalyze the NAD ϩ -dependent oxidation of a spontaneously formed product of mycothiol and formaldehyde, S-hydroxymethylmycothiol, to S-formyl-mycothiol, which can subsequently be hydrolyzed to formate and mycothiol (48)(49)(50).…”

Section: Oe) (B)mentioning

confidence: 99%

“…The latter possibility appears unlikely for C. glutamicum, since the ⌬fdhF mutant converted methanol stoichiometrically to formate. The gene located immediately downstream of adhE in C. glutamicum, cg0388, is annotated as Zn-dependent hydrolase, and the derived protein sequence has 65% sequence identity to that of the proposed R. erythropolis FMH (47). In addition, the putative lysophospholipase (cg2193) shows 38% sequence identity to the Mycobacterium tuberculosis Rv2854 protein, which is expected to be an FMH.…”

Section: Oe) (B)mentioning

confidence: 99%

See 2 more Smart Citations

C ₁ Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide

Witthoff

Marienhagen

Bott

2013

Appl Environ Microbiol

View full text Add to dashboard Cite

Methanol is considered an interesting carbon source in "bio-based" microbial production processes. Since Corynebacterium glutamicum is an important host in industrial biotechnology, in particular for amino acid production, we performed studies of the response of this organism to methanol. The C. glutamicum wild type was able to convert 13 C-labeled methanol to 13 CO 2 . Analysis of global gene expression in the presence of methanol revealed several genes of ethanol catabolism to be upregulated, indicating that some of the corresponding enzymes are involved in methanol oxidation. Indeed, a mutant lacking the alcohol dehydrogenase gene adhA showed a 62% reduced methanol consumption rate, indicating that AdhA is mainly responsible for methanol oxidation to formaldehyde. Further studies revealed that oxidation of formaldehyde to formate is catalyzed predominantly by two enzymes, the acetaldehyde dehydrogenase Ald and the mycothiol-dependent formaldehyde dehydrogenase AdhE. The ⌬ald ⌬adhE and ⌬ald ⌬mshC deletion mutants were severely impaired in their ability to oxidize formaldehyde, but residual methanol oxidation to CO 2 was still possible. The oxidation of formate to CO 2 is catalyzed by the formate dehydrogenase FdhF, recently identified by us. Similar to the case with ethanol, methanol catabolism is subject to carbon catabolite repression in the presence of glucose and is dependent on the transcriptional regulator RamA, which was previously shown to be essential for expression of adhA and ald. In conclusion, we were able to show that C. glutamicum possesses an endogenous pathway for methanol oxidation to CO 2 and to identify the enzymes and a transcriptional regulator involved in this pathway.

show abstract

Section: Discussionmentioning

confidence: 65%

Section: Oe) (B)mentioning

confidence: 99%

Section: Oe) (B)mentioning

confidence: 99%

See 1 more Smart Citation

C ₁ Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide

Witthoff

Marienhagen

Bott

2013

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Glyoxalase II performs a reaction similar to that proposed for FmhA: the hydrolysis of thioester S-D-lactoyl-glutathione to D-lactate and glutathione (57). Interestingly, adhE, fmhA, fdhD, and fdhF are not clustered on either the C. glutamicum genome or the Rhodococcus erythropolis genome (55,58).…”

Section: Discussionmentioning

confidence: 93%

Characterization of p -Hydroxycinnamate Catabolism in a Soil Actinobacterium

et al. 2014

View full text Add to dashboard Cite

p-Hydroxycinnamates, such as ferulate and p-coumarate, are components of plant cell walls and have a number of commercial applications. Rhodococcus jostii RHA1 (RHA1) catabolizes ferulate via vanillate and the ␤-ketoadipate pathway. Here, we used transcriptomics to identify genes in RHA1 that are upregulated during growth on ferulate versus benzoate. The upregulated genes included three transcriptional units predicted to encode the uptake and ␤-oxidative deacetylation of p-hydroxycinnamates: couHTL, couNOM, and couR. Neither ⌬couL mutants nor ⌬couO mutants grew on p-hydroxycinnamates, but they did grow on vanillate. Among several p-hydroxycinnamates, CouL catalyzed the thioesterification of p-coumarate and caffeate most efficiently (k cat /K m ‫؍‬ ϳ400 mM ؊1 s ؊1 ). p-Coumarate was also RHA1's preferred growth substrate, suggesting that CouL is a determinant of the pathway's specificity. CouL did not catalyze the activation of sinapate, in similarity to two p-coumaric acid:coenzyme A (CoA) ligases from plants, and contains the same bulged loop that helps determine substrate specificity in the plant homologues. The couO mutant accumulated 4-hydroxy-3-methoxyphenyl-␤-ketopropionate in the culture supernatant when incubated with ferulate, supporting ␤-oxidative deacetylation. This phenotype was not complemented with a D257N variant of CouO, consistent with the predicted role of Asp257 as a metal ligand in this amidohydrolase superfamily member. These data suggest that CouO functionally replaces the ␤-ketothiolase and acyl-CoA thioesterase that occur in canonical ␤-oxidative pathways. Finally, the transcriptomics data suggest the involvement of two distinct formaldehyde detoxification pathways in vanillate catabolism and identify a eugenol catabolic pathway. The results of this study augment our understanding of the bacterial catabolism of aromatics from renewable feedstocks.

show abstract

“…Proteomic and transcriptomic analyses showed that two enzymes involved in the methanol oxidation of Gram-positive bacteria, N,N'-dimethyl-4-nitrosoaniline (NDMA)-dependent methanol dehydrogenase (MDH) and NAD-dependent formaldehyde dehydrogenase (nFADH), were highly expressed in N9T-4 under oligotrophic conditions Yoshida et al 2011). It is known that MDH has a remarkable activity for formaldehyde dismutase in addition to methanol dehydrogenase (Bystrykh et al 1993;Ohhata et al 2007).…”

Section: Introductionmentioning

confidence: 99%

The glyoxylate shunt is essential for CO2-requiring oligotrophic growth of Rhodococcus erythropolis N9T-4

Yokota

Yoshida

et al. 2015

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

Rhodococcus erythropolis N9T-4 shows extremely oligotrophic growth requiring atmospheric CO2 and forms its colonies on an inorganic basal medium (BM) without any additional carbon source. Screening of a random mutation library constructed by a unique genome deletion method that we established indicated that the aceA, aceB, and pckG genes encoding isocitrate lyase, malate synthase, and phosphoenolpyruvate carboxykinase, respectively, were requisite for survival on BM plates. The aceA- and aceB deletion mutants and the pckG deletion mutant grew well on BM plates containing L-malate and D-glucose, respectively, suggesting that the glyoxylate (GO) shunt and gluconeogenesis are essential for the oligotrophic growth of N9T-4. Interestingly, most of the enzyme activities in the TCA cycle were observed in the cell-free extract of N9T-4, with perhaps the most important exception being α-ketoglutarate dehydrogenase (KGDH) activity. Instead of the KGDH activity, we detected a remarkable level of α-ketoglutarate decarboxylase (KGD) activity, which is the activity exhibited by the E1 component of the KGDH complex in Mycobacterium tuberculosis. The recombinant KGD of N9T-4 catalyzed the decarboxylation of α-ketoglutarate to form succinic semialdehyde (SSA) in a time-dependent manner. Since N9T-4 also showed a detectable SSA dehydrogenase activity, we concluded that N9T-4 possesses a variant TCA cycle, which uses SSA rather than succinyl-CoA. These results suggest that oligotrophic N9T-4 cells utilize the GO shunt to avoid the loss of carbons as CO2 and to conserve CoA units in the TCA cycle.

show abstract

Gene Expression Analysis of Methylotrophic Oxidoreductases Involved in the Oligotrophic Growth ofRhodococcus erythropolisN9T-4

Cited by 21 publications

References 10 publications

C ₁ Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide

C ₁ Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide

Characterization of p -Hydroxycinnamate Catabolism in a Soil Actinobacterium

The glyoxylate shunt is essential for CO2-requiring oligotrophic growth of Rhodococcus erythropolis N9T-4

Contact Info

Product

Resources

About

Gene Expression Analysis of Methylotrophic Oxidoreductases Involved in the Oligotrophic Growth ofRhodococcus erythropolisN9T-4

Cited by 21 publications

References 10 publications

C 1 Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide

C 1 Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide

Characterization of p -Hydroxycinnamate Catabolism in a Soil Actinobacterium

The glyoxylate shunt is essential for CO2-requiring oligotrophic growth of Rhodococcus erythropolis N9T-4

Contact Info

Product

Resources

About

C ₁ Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide

C ₁ Metabolism in Corynebacterium glutamicum: an Endogenous Pathway for Oxidation of Methanol to Carbon Dioxide