Reductase component of the toluene-4-monooxygenase multicomponent enzyme system which catalyzes the O2- and NADH-dependent hydroxylation of toluene to form p-cresol (PubMed:15240250, PubMed:17964805). Ferredoxin reductase catalyzes the transfer of electrons from NADH to ferredoxin (TmoC). H+ + NAD+ + 2 reduced [2Fe-2S]-[ferredoxin] = NADH + 2 oxidized [2Fe-2S]-[ferredoxin]
MTDTRHSTGATGAPGTGTTGSAGATGDAAPRRTTRPMTGDEYLESIRDGREIWAYGERVD
DVTKHPAFRNTARMTARLYDALHDPEHHDTLTTPTDTGSDGYTHKFYRVPRSVEDLVGDR
DAIACWARMTYGWMGRSPDYKASFLVTLGADPDYYGEFADNARRWYAEAQERVLFWNHAV
INPPVDRNRAPDEVGDVFVHVEKECDDGLVVSGAKVVATGSALTHFNFVAHYGLPVKKKE
FALVATLPMDAPGVKLICRQSYELAANRMGSPFDYPLSSRLDENDTVFILDKVKIPWENV
FIYGDTAKAGTFLNTSGFTHRLTFHGVTRLAVKLDFLAGLLLKGLDVTGTKDFRGIQTRV
GEVLAWRNMFWGLSDAMAHNPNAWHDGGLLPNLDYGMAYRWFMTIGYPRVREIILQDLSS
GLIYLNSHAKDFQNPELRPHLDRYMRGSNGYDSVERVKLMKLIWDSVGTEFGGRHELYER
NYSGNHENVRIELLMAQTASGLVDGYRGFAEECMSEYDLDGWTAPDLIGPDQA541
| PMID | Title & Author | Abstract | Year | |
| 0 | 28109950 | Cloning, genetic engineering and characterization of TMOF expressed in Saccharomyces cerevisiae to control larval mosquitoes Dov Borovsky , Sabine Nauewelaers , Charles A Powell , Robert G Shatters Jr | Trypsin modulating oostatic factor, a decapaptide isolated from the ovaries of A. aegypti, is the physiological factor that terminates the trypsin biosynthesis after the blood meal. Earlier results obtained from feeding mosquito larvae and injecting female mosquitoes with TMOF show that trypsin biosynthesis and egg development are inhibited, indicating that TMOF traverses the gut epithelial cells and modulates trypsin biosynthesis, making it a potential larvacidal peptide hormone. Therefore, TMOF and TMOF green fluorescent protein (GFP) fusion protein with a trypsin cleavage site, allowing TMOF release in the larval gut, were expressed in S. cerevisiae cells that were transformed using homologous recombination at ura3-52 with an engineered plasmid (pYDB2) carrying tmfA and gfp-tmfA and a strong galactose promoter (PGAL1). Southern blot analyses showed that each cell incorporated a single tmfA or gfp-tmfA. Western blot analyses of cells that were fermented up to 48h showed that the engineered S. cerevisiae cells synthesized both TMOF and GFP-TMOF and heat treatment did not affect the recombinant proteins. Engineered S. cerevisiae (3×108cells) that were fermented for 4h produced (2.1±0.2μg±S.E.M) of TMOF. Feeding the engineered cells producing TMOF and GFP-TMOF to larval mosquito caused high mortalities (66±12% and 83±8%, respectively). S. cerevisiae cells transfected with pYEX-BX carrying gfp-tmfA and (DPAR)4 or transformed by homologous recombination of pYDB2-gfp-tmfA carrying a heat shock promoter (PHP) were ineffective. Engineered heat treated yeast cells are consumed by mosquito larvae, and could be used to control mosquitoes. | 2017 |
| 1 | 1429451 | Identification of a new gene, tmoF, in the Pseudomonas mendocina KR1 gene cluster encoding toluene-4-monooxygenase K M Yen , M R Karl | Five genes, tmoABCDE, encoding toluene-4-monooxygenase (T4MO) were previously mapped to a 3.6-kb region of a 10.2-kb SacI DNA fragment isolated from Pseudomonas mendocina KR1 (K.-M. Yen, M. R. Karl, L. M. Blatt, M. J. Simon, R. B. Winter, P. R. Fausset, H. S. Lu, A. A. Harcourt, and K. K. Chen, J. Bacteriol. 173:5315-5327, 1991). In this report, we describe the identification and characterization of a DNA region in the SacI fragment whose expression enhances the T4MO activity determined by the tmoABCDE gene cluster. This region was mapped immediately downstream of the putative transcription termination sequence previously located at the end of the tmoABCDE gene cluster (Yen et al., J. Bacteriol., 1991) and was found to stimulate T4MO activity two- to threefold when expressed in Escherichia coli or Pseudomonas putida. Determination of the nucleotide sequence of this region revealed an open reading frame (ORF) of 978 bp. Expression of the ORF resulted in the synthesis of an approximately 37-kDa polypeptide whose N-terminal amino acid sequence completely matched that of the product predicted from the ORF. The ORF thus defines a gene, which has now been designated tmoF. The TmoF protein shares amino acid sequence homology with the reductases of several mono- and dioxygenase systems. In addition, the reductase component of the naphthalene dioxygenase system, encoded by the nahAa gene of plasmid NAH7 from P. putida G7, could largely replace the TmoF protein in stimulating T4MO activity, and TmoF could partially replace the NahAa protein in forming active naphthalene dioxygenase. The overall properties of tmoF suggest that it is a member of the T4mo gene cluster and encodes the NADH:ferredoxin oxidoreductase of the T4MO system. | 1992 |
Yen KM, Karl MR. Identification of a new gene, tmoF, in the Pseudomonas mendocina KR1 gene cluster encoding toluene-4-monooxygenase.(J) Bacteriol 174:7253-61 (1992)