Expression of Double <i>Vitreoscilla</i> Hemoglobin Enhances Growth and Alters Ribosome and tRNA Levels in <i>Escherichia coli</i>

Roos, Viktoria; Andersson, Charlotte I.J.; Arfvidsson, Cecilia; Wahlund, Karl-Gustav; Bülow, Leif

doi:10.1021/bp020005v

Cited by 17 publications

(10 citation statements)

References 23 publications

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“…By site-directed mutagenesis to remove the loop responsible for the intersubunit interaction, Piubelli et al (2002) converted the dimeric RtDAO into a monomeric form and showed that dimerization was required for a tighter FAD binding and higher thermostability. Several previous studies demonstrated that the subunit fusion of several dimeric proteins into single-chain variants by genetic manipulation could not only improve their stability but also increase the catalytic activity or enhance protein physiological functions (Liang et al 1993;Robinson and Sauer 1996;Roos et al 2002). In this paper, we report that the covalent linking of subunits of RtDAO and TvDAO resulted in significant improvement in thermal stability and tolerance against oxidative damage by hydrogen peroxide.…”

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confidence: 78%

Subunit fusion of two yeast d-amino acid oxidases enhances their thermostability and resistance to H2O2

Wang

Lee

et al. 2008

Biotechnol Lett

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D-amino acid oxidases from Rhodosporidium toruloides and Trigonopsis variabilis (RtDAO and TvDAO) are both yeast homodimeric flavoenzymes. Two of their cDNA genes were connected by a hexanucleotide linker and heterologously expressed in E. coli to produce the corresponding double DAOs (dRtDAO and dTvDAO) with two subunits fused into a single polypeptide. The specific activities of double DAOs remained similar to those of native dimeric DAOs, although the catalytic efficiencies (k(cat)/K(M)) were decreased due to higher K(M) values. The T(m) value for dRtDAO was shifted 5 degrees C higher while that for dTvDAO was increased only by 2 degrees C, in comparison with the corresponding native counterparts. In the presence of 10 mM H(2)O(2), dRtDAO and dTvDAO exhibited half-lives of about 60 and 40 min, respectively, which were 2- and 1.5-fold, respectively, longer than their native DAOs. These yeast DAOs can therefore be thermally and oxidatively stabilized by linking their subunits together.

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confidence: 78%

Subunit fusion of two yeast d-amino acid oxidases enhances their thermostability and resistance to H2O2

Wang

Lee

et al. 2008

Biotechnol Lett

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“…A reductase domain from the Fhp (R. eutropha) flavohemoglobin was genetically linked to Vgb with an improvement in the beneficial effects on microaerobic bacterial growth (15,23,27). Expression of a double Vgb-Vgb hemoglobin also enhanced growth and alters ribosome and tRNA levels in E. coli (49).…”

Section: Discussionmentioning

confidence: 99%

Flavohemoglobin Hmp, but Not Its Individual Domains, Confers Protection from Respiratory Inhibition by Nitric Oxide in Escherichia coli

Hernández-Urzúa¹,

Mills

White

et al. 2003

Journal of Biological Chemistry

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Escherichia coli possesses a two-domain flavohemoglobin, Hmp, implicated in nitric oxide (NO) detoxification. To determine the contribution of each domain of Hmp toward NO detoxification, we genetically engineered the Hmp protein and separately expressed the heme (HD) and the flavin (FD) domains in a defined hmp mutant. Expression of each domain was confirmed by Western blot analysis. CO-difference spectra showed that the HD of Hmp can bind CO, but the CO adduct showed a slightly blue-shifted peak. Overexpression of the HD resulted in an improvement of growth to a similar extent to that observed with the Vitreoscilla hemeonly globin Vgb, whereas the FD alone did not improve growth. Viability of the hmp mutant in the presence of lethal concentrations of sodium nitroprusside was increased (to 30% survival after 2 h in 5 mM sodium nitroprusside) by overexpressing Vgb or the HD. However, maximal protection was provided only by holo-Hmp (75% survival under the same conditions). Cellular respiration of the hmp mutant was instantaneously inhibited in the presence of 13.5 M NO but remained insensitive to NO inhibition when these cells overexpressed Hmp. When HD or FD was expressed separately, no significant protection was observed. By contrast, overexpression of Vgb provided partial protection from NO respiratory inhibition. Our results suggest that, despite the homology between the HD from Hmp and Vgb (45% identity), their roles seem to be quite distinct.

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“…In a number of studies (Liu et al. 1995; Roos et al. 2002; Bhave and Chattoo 2003; Suthar and Chattoo 2006; Liang et al.…”

Section: Discussionmentioning

confidence: 99%

“…In a number of studies (Liu et al 1995;Roos et al 2002;Bhave and Chattoo 2003;Suthar and Chattoo 2006;Liang et al 2007), improvement of growth properties had been detected in various hosts with exogenous expression of VHb. To investigate whether the strain improvement in this study was because of the expression of the vgb gene, randomly chosen transformants were tested and compared with the two control groups (randomly selected strains bearing empty vectors and WT strain).…”

Section: Discussionmentioning

confidence: 99%

Constitutive expression of Vitreoscilla haemoglobin in Sphingomonas elodea to improve gellan gum production

Chen

et al. 2010

Journal of Applied Microbiology

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Aims: To improve a commercially used strain for gellan production by exogenous Vitreoscilla haemoglobin (VHb). Methods and Results: VHb gene was expressed in Sphingomonas elodea under the control of constitutive bla promoter. Biochemical activity of expressed VHb was confirmed by CO‐difference spectra analysis that exhibited a characteristic absorption maximum at 419 nm. During cultivation, not only enhanced cell growth was detected, but also 20% improvement in gellan production was observed after 48 h of incubation, with a maximum yield of 16·82 g l−1. Moreover, maximum sucrose conversion efficiency (g gellan per g sucrose) was 57·8, 20% higher than that of the parental strain. We further examined the polysaccharide production of VHb‐expressing strain at different aeration levels in Erlenmeyer flasks. Again, in all cases, a significant enhancement of gellan production was observed, and the enhancement was more significant under oxygen‐limiting conditions (up to 26·8%). Conclusions: VHb exhibited positive effect on cell growth and gellan yield of S. elodea, especially under hypoxic conditions. Significance and Impact of the Study: This is the first application of VHb as an effective metabolic engineering strategy in S. elodea to regulate cell growth and optimize gellan yield.

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Expression of Double Vitreoscilla Hemoglobin Enhances Growth and Alters Ribosome and tRNA Levels in Escherichia coli

Cited by 17 publications

References 23 publications

Subunit fusion of two yeast d-amino acid oxidases enhances their thermostability and resistance to H2O2

Subunit fusion of two yeast d-amino acid oxidases enhances their thermostability and resistance to H2O2

Flavohemoglobin Hmp, but Not Its Individual Domains, Confers Protection from Respiratory Inhibition by Nitric Oxide in Escherichia coli

Constitutive expression of Vitreoscilla haemoglobin in Sphingomonas elodea to improve gellan gum production

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