Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:Q8NEX9 (reductase)
 26,410 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The fatty acid synthetase (FAS) gene FAS1 of the alkane-utilizing yeast Yarrowia lipolytica was cloned and sequenced. The gene is represented by an intron-free reading frame of 6228 bp encoding a protein of 2076 amino acids and 229,980 Da molecular weight. This protein exhibits a 58% sequence similarity to the corresponding Saccharomyces cerevisiae FAS beta-subunit. The sequential order of the five FAS1-encoded enzyme The sequential order of the five FAS1-encoded enzyme domains, acetyl transferase, enoyl reductase, dehydratase and malonyl/palmityl-transferase, is co-linear in both organisms. This finding agrees with available evidence that the functional organization of FAS genes is similar in related organisms but differs considerably between unrelated species. In addition, previously reported conflicting data concerning the 3' end of S. cerevisiae FAS1 were re-examined by genomic and cDNA sequencing of the relevant portion of the gene. Thereby, the translational stop codon was shown to lie considerably downstream of both published termination sites. The S. cerevisiae FAS1 gene thus has a corrected length of 6153 bp and encodes a protein of 2051 amino acids and 228,667 Da molecular weight.
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PMID:The pentafunctional FAS1 genes of Saccharomyces cerevisiae and Yarrowia lipolytica are co-linear and considerably longer than previously estimated. 203 24

The yeast fatty acid synthase is a complex (alpha 6 beta 6) of two multifunctional proteins alpha and beta. The alpha subunit (Mr 212,000) contains two of the seven enzymatic activities required for the synthesis of fatty acids and the site for attachment of the prosthetic group 4'-phosphopantetheine. The beta subunit (Mr 203,000) contains the remaining five activities. Cloning of the genes encoding the alpha and beta proteins has been reported (Kuziora, M. A., Chalmers, J. H., Jr., Hitzeman, R. A., Douglas, M. G., and Wakil, S. J. (1983) J. Biol. Chem. 258, 11648-11653). In the present study it is shown that two of the clones containing the beta subunit gene, YEpFAS1 and YEp33F1, are not identical. The clone YEp33F1 contains the gene that codes for the entire beta subunit while YEpFAS1 is missing approximately half of the gene at the 3' end. Despite this loss, YEpFAS1 is still able to complement a fas1 mutation at the enoyl reductase domain. This complementation does not occur by recombination, rather a small mRNA is produced in cells transformed with YEpFAS1 and is translated into a protein of molecular weight of approximately 125,000 which is immunologically reactive with yeast fatty acid synthase antibodies. The data suggest that this truncated beta subunit interacts with the mutant alpha 6 beta 6 complex to restore fatty acid synthesis to the cell. The nucleotide sequence of the FAS1 gene cloned in YEp33F1 DNA, which encodes the beta subunit of fatty acid synthase, was determined. The coding region consists of 5940 base pairs (bp) and could encode a protein of 1980 amino acids with a calculated molecular weight of 220,077. A major transcriptional start point was mapped to a position of about 330 bp upstream from the first ATG codon. The termination of transcription was mapped at about 300 bp downstream from the first TGA stop codon. The sequence of the beta subunit protein does not appear to be similar to any other sequenced protein. The sites of the active seryl groups for the acetyltransacylase and malonyl/palmitoyl transacylase were identified from known amino acid sequences to be residues 274 and 1808, respectively. Putative binding sites for FMN and NADPH were suggested based on similarities with amino acid sequences of known flavin and pyridine nucleotide enzymes, respectively.
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PMID:Complementation of mutations and nucleotide sequence of FAS1 gene encoding beta subunit of yeast fatty acid synthase. 303 Oct 66

Yarrowia lipolytica, like other lower fungi, has a fatty acid synthetase complex (FAS) with an alpha 6 beta 6 molecular structure. Both subunits are multifunctional proteins each with a molecular weight of more then 200,000 daltons. A collection of FAS-deficient) Y. lipolytica mutants was isolated and characterized by both genetic complementation and enzyme activity measurements. It was found that the three acyl transferases (acetyl-, malonyl- and palmityl-transacylation) together with the enoyl reductase domain are located on subunit beta and, therefore, are encoded by the gene locus FAS1. beta-Ketoacyl reductase, beta-ketoacyl synthase and acyl carrier protein functions are part of the FAS2-encoded subunit alpha. Thus, the functional organization of FAS1 and FAS2 is identical in both yeasts, Saccharomyces cerevisiae and Yarrowia lipolytica. Nevertheless, the two yeasts differ significantly with respect to the intragenic complementation characteristics of fas1 and fas2 mutants. This finding is discussed in terms of a specific inter- or intramolecular reaction mechanism within the oligomeric FAS complex. The pentafunctional Y. lipolytica FAS1 gene was isolated from a lambda gt11 expression library using polyclonal antisera against the purified FAS complex. At present, sequencing of FAS1, which is more than 5 kilobases long, is almost completed. Available data indicate approx. 60 percent sequence homology together with an identical order of catalytic domains within subunit beta of the two yeasts, Y. lipolytica and S. cerevisiae.
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PMID:Genetic control of Yarrowia lipolytica fatty acid synthetase biosynthesis and function. 323 May 12

FAS1, the structural gene of the pentafunctional fatty acid synthetase subunit beta in Saccharomyces cerevisiae has been sequenced. Its reading frame represents an intron-free nucleotide sequence of 5,535 base pairs, corresponding to a protein of 1,845 amino acids with a molecular weight of 205,130 daltons. In addition to the coding sequence, 1,468 base pairs of its 5'-flanking region were determined. S1 nuclease mapping revealed two transcriptional initiation sites; 5 and 36 base pairs upstream of the translational start codon. Within the flanking sequences two TATATAAA boxes, several A-rich and T-rich blocks and a TAG...TATGTT...TATGTT...TTT sequence were found and are discussed as transcriptional initiation and termination signals, respectively. The order of catalytic domains in the cluster gene was established by complementation of defined fas1 mutants with overlapping FAS1 subclones. Acetyl transferase (amino acids 1-468) is located proximal to the N-terminus of subunit beta, followed by the enoyl reductase (amino acids 480-858), the dehydratase (amino acids 1,134-1,615) and the malonyl/palmityl transferase (amino acids 1,616-1,845) domains. One major inter-domain region of about 276 amino acids with so far unknown function was found between the enoyl reductase and dehydratase domains. The substrate-binding serine residues of acetyl, malonyl and palmityl transferases were identified within the corresponding domains. Significant sequence homologies exist between the acyl transferase active sites of yeast and animal fatty acid synthetases. Similarly, a putative sequence of the enoyl reductase active site was identified.
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PMID:The pentafunctional FAS1 gene of yeast: its nucleotide sequence and order of the catalytic domains. 352 50

From a Saccharomyces cerevisiae gene bank contained in the novel yeast cosmid shuttle vector pMS201 the fatty acid synthetase (FAS) genes FAS1 and FAS2 were isolated. FAS clones were identified by in situ colony hybridization using two yeast DNA probes apparently capable of producing avian FAS cross-reacting material (J. Carbon, personal communication). Classification as FAS1 or FAS2 clones was achieved by their specific transformation of fas1 and fas2 yeast mutants. By transcription mapping FAS1 was assigned to about 5.3 kb within 14.8 kb of chromosomal DNA covered by two genomically adjacent BamHI fragments. The FAS2 gene was localized on a single BamHI fragment of 25 kb. One of the FAS clones ( FAS2 ) produces immunologically cross-reacting material in Escherichia coli. High frequency transformation of fas1 mutants was only observed with one subclone, pMS3021 , containing the intact FAS1 locus. Other DNA segments cloned in the same self-replicating vector but representing only part of FAS1 exhibited drastically lower transformation rates. As evident from this and from FAS1 /TRP1-cotransformation rates only the intact FAS1 gene in pMS3021 is capable of fas1 -mutant complementation. With partial FAS1 genes, even when coding for an intact equivalent of the mutated domain, their chromosomal integration is necessary for the expression of FAS. In integrative transformants the coexistence of integrated and autonomously replicating plasmid DNA was demonstrated. Both, the extrachromosomal and chromosomally integrated FAS DNA was mitotically unstable. Transformation studies using subcloned FAS1 DNA segments revealed the relative locations of the enoyl reductase and dehydratase domains within this pentafunctional cluster gene.
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PMID:Molecular cloning of the yeast fatty acid synthetase genes, FAS1 and FAS2: illustrating the structure of the FAS1 cluster gene by transcript mapping and transformation studies. 633 May 2

A gene encoding a fatty acid synthase component, FAS1, has been cloned from a genomic library of the polyunsaturated fatty acid (PUFA)-producing yeast Saccharomyces kluyveri. This gene (named Sk-FAS1) was found to contain an open reading frame of 6150 bp, coding for 2049 amino acids. The deduced Sk-FAS1 protein showed significant (75-59%) homology with FAS proteins from the other yeasts, including S. cerevisiae, Candida albicans and Yarrowia lipolytica. The substrate-binding sites of the acetyl transferase and malonyl/palmitoyl transferase domains, and the FMN- and NADPH-binding sites of the enoyl reductase domain, were all highly conserved. Expression of the Sk-FAS1 gene in S. cerevisiae complemented genetic disruption of the S. cerevisiae FAS1 gene (Sc-FAS1), suggesting the formation of a heterogeneous complex of Sk-FAS1 (beta) and Sc-FAS2 (alpha), which is able to function to synthesize fatty acids. Compared with the isogenic wild-type of S. cerevisiae, as well as S. kluyveri, the S. cerevisiae fas1 mutant carrying the Sk-FAS1 gene showed an increase in the relative amount of 16-carbon fatty acids and a decrease in 18-carbon fatty acids.
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PMID:Cloning of a fatty acid synthase component FAS1 gene from Saccharomyces kluyveri and its functional complementation of S. cerevisiae fas1 mutant. 1157 58

Here we report recombinant expression and activity of several type I fatty acid synthases that can function in parallel with the native Escherichia coli fatty acid synthase. Corynebacterium glutamicum FAS1A was the most active in E. coli and this fatty acid synthase was leveraged to produce oleochemicals including fatty alcohols and methyl ketones. Coexpression of FAS1A with the ACP/CoA-reductase Maqu2220 from Marinobacter aquaeolei shifted the chain length distribution of fatty alcohols produced. Coexpression of FAS1A with FadM, FadB, and an acyl-CoA-oxidase from Micrococcus luteus resulted in the production of methyl ketones, although at a lower level than cells using the native FAS. This work, to our knowledge, is the first example of in vivo function of a heterologous fatty acid synthase in E. coli. Using FAS1 enzymes for oleochemical production have several potential advantages, and further optimization of this system could lead to strains with more efficient conversion to desired products. Finally, functional expression of these large enzyme complexes in E. coli will enable their study without culturing the native organisms.
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PMID:Development of an orthogonal fatty acid biosynthesis system in E. coli for oleochemical production. 2588 38