Bioconversion of tyrosine into the propylhygric acid moiety of lincomycin

Witz, D. F.; Hessler, Edward J.; Miller, Thomas Lee

doi:10.1021/bi00783a005

Cited by 36 publications

(14 citation statements)

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“…M. luteus was cultured at 37°C in medium III (5 g/liter polypeptone, 1.5 g/liter beef extract, 3 g/liter yeast extract, 3.5 g/liter NaCl, 3.68 g/liter K 2 HPO 4 , 1.32 g/liter KH 2 PO 4 , 1 g/liter glucose, 18 g/liter agar) for 16 to 18 h. The cell pellets were washed with sterile 0.9% NaCl solution to prepare the M. luteus culture suspension. Lincomycin standard solutions (2,4,6,8,10, and 12 g/ml) were used to prepare the standard curves. Inhibition zone diameters were linear with the logarithmic values of the concentrations of the lincomycin standard solutions.…”

Section: Methodsmentioning

confidence: 99%

“…L-Tyrosine was identified as a precursor of PPL through stable-isotope labeling (10), and it is converted into L-3,4-dihydroxyphenylalanine (L-DOPA) and then into 4-(3-carboxy-3-oxo-propenyl)-2,3-dihydro-1H-pyrrole-2-carboxylic acid (product 1) by the L-tyrosine hydroxylase LmbB2 and the L-DOPA-2,3-dioxygenase LmbB1 (11,12). In the recent report, it was proposed that product 1 undergoes methylation by LmbW to generate product 2 (13,14).…”

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confidence: 99%

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The Novel Transcriptional Regulator LmbU Promotes Lincomycin Biosynthesis through Regulating Expression of Its Target Genes in Streptomyces lincolnensis

Hou

Lin

et al. 2018

J Bacteriol

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Lincomycin A is a clinically important antimicrobial agent produced by Streptomyces lincolnensis. In this study, a new regulator designated LmbU (GenBank accession no. ABX00623.1) was identified and characterized to regulate lincomycin biosynthesis in S. lincolnensis wild-type strain NRRL 2936. Both inactivation and overexpression of lmbU resulted in significant influences on lincomycin production. Transcriptional analysis and in vivo neomycin resistance (Neo r ) reporter assays demonstrated that LmbU activates expression of the lmbA, lmbC, lmbJ, and lmbW genes and represses expression of the lmbK and lmbU genes. Electrophoretic mobility shift assays (EMSAs) demonstrated that LmbU can bind to the regions upstream of the lmbA and lmbW genes through the consensus and palindromic sequence 5=-CGCCG GCG-3=. However, LmbU cannot bind to the regions upstream of the lmbC, lmbJ, lmbK, and lmbU genes as they lack this motif. These data indicate a complex transcriptional regulatory mechanism of LmbU. LmbU homologues are present in the biosynthetic gene clusters of secondary metabolites of many other actinomycetes. Furthermore, the LmbU homologue from Saccharopolyspora erythraea (GenBank accession no. WP_009944629.1) also binds to the regions upstream of lmbA and lmbW, which suggests widespread activity for this regulator. LmbU homologues have no significant structural similarities to other known cluster-situated regulators (CSRs), which indicates that they belong to a new family of regulatory proteins. In conclusion, the present report identifies LmbU as a novel transcriptional regulator and provides new insights into regulation of lincomycin biosynthesis in S. lincolnensis. IMPORTANCE Although lincomycin biosynthesis has been extensively studied, its regulatory mechanism remains elusive. Here, a novel regulator, LmbU, which regulates transcription of its target genes in the lincomycin biosynthetic gene cluster (lmb gene cluster) and therefore promotes lincomycin biosynthesis, was identified in S. lincolnensis strain NRRL 2936. Importantly, we show that this new regulatory element is relatively widespread across diverse actinomycetes species. In addition, our findings provide a new strategy for improvement of yield of lincomycin through manipulation of LmbU, and this approach could also be evaluated in other secondary metabolite gene clusters containing this regulatory protein.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

The Novel Transcriptional Regulator LmbU Promotes Lincomycin Biosynthesis through Regulating Expression of Its Target Genes in Streptomyces lincolnensis

Hou

Lin

et al. 2018

J Bacteriol

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“…) The biosynthesis of the hydropyrrole moiety of anthramycin, sibiromycin, and tomaymycin proceed through a common pathway also shared by the biosynthesis of lincomycin, a natural product produced by Streptomyces lincolnensis , (Figs. and ).…”

Section: Biosynthesis Of the Pyrrolobenzodiazepinesmentioning

confidence: 99%

Biosynthesis, synthesis, and biological activities of pyrrolobenzodiazepines

Gerratana

2010

Medicinal Research Reviews

118

143

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Pyrrolobenzodiazepines (PBDs) are sequence selective DNA alkylating agents with remarkable antineoplastic activity. They are either naturally produced by actinomycetes or synthetically produced. The remarkable broad spectrum of activities of the naturally produced PBDs encouraged the synthesis of several PBDs, including dimeric and hybrid PBDs yielding to an improvement in the DNA binding sequence specificity and in the potency of this class of compounds. However, limitation in the chemical synthesis prevented the testing of one of the most potent PBDs, sibiromycin, a naturally produced glycosylated PBDs. Only recently the biosynthetic gene clusters for PBDs have been identified opening the doors to the production of glycosylated PBDs by mutasynthesis and biosynthetic engineering. The present review describes the recent studies on the biosynthesis of naturally produced pyrrolobenzodiazepines. In addition, it provides an overview on the isolation and characterization of naturally produced PBDs, on the chemical synthesis of PBDs, on the mechanism of DNA alkylation, and on the DNA binding affinity and cytotoxic properties of both naturally produced and synthetic pyrrolobenzodiazepines.

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“…Recent results on the biogenesis of lincomycin suggested that these enzymes may be involved in the synthesis of lincomycin by Streptomyces lincolnensis (19). The isotopic experiments performed with this organism showed that the pyrrolidone structure of the propylhydric moiety of lincomycin originates from L-tyrosine.…”

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confidence: 95%

Monophenol Monooxygenase and Lincomysin Biosynthesis in Streptomyces lincolnensis

Michalik

Emilianowicz-Czerska

Switalski

et al. 1975

Antimicrob Agents Chemother

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Monophenol monooxygenase (monophenol, dihydroxyphenylalanine:oxygen oxidoreductase EC 1.14.18.1) was studied in melanin-positive and melaninnegative mutants ofStreptomyces lincolnensis NCIB 9413, varying in the lincomycin synthesizing ability. The activities of laccase and tyrosine phenol lyase (EC 4.1.99.2) are absent in this organism. The monophenol monooxygenase catalyzes hydroxylation of monophenols (K. and V., for L-tyrosine, 2 x 1(4 M and 8.0 nmol of O2/min per ml, respectively) at a slower rate than it dehydrogenates diphenols to o-quinones (Km and Vmax for L-3,4-dihydroxyphenylalanine, 7 x 10-5 M and 51.7 nmol of 0/min per ml, respectively. It is inhibited by KCN, /3-mercaptoethanol, ethylenediaminetetraacetate, dipyridyl, thiourea, p-aminobenzoic acids and by some tryptophan metabolites. Changes in the activity of monophenol monooxygenase caused by mutation or by inhibitors are reflected in the synthesis ofthe antibiotic. Its participation in the biogenesis ofthe propylhygric moiety of lincomycin is discussed.It has been reported that phenol oxidase enzymes participate in the formation of fruiting bodies in the basidiomycete Schizophyllum commune (12). Recent results on the biogenesis of lincomycin suggested that these enzymes may be involved in the synthesis of lincomycin by Streptomyces lincolnensis (19). The isotopic experiments performed with this organism showed that the pyrrolidone structure of the propylhydric moiety of lincomycin originates from L-tyrosine. According to these data propylproline or ethylproline, the respective precursors of propyl and ethylhygric acids, are formed from L-dopa quinone or 2-carboxy-2,3-dihydro-5,6-dihydroxyindole. The latter two compounds are also intermediates of melanogenesis. This may imply that monophenol monooxygenase is the enzyme initiating both the formation of melanin and the biosynthesis of lincomycin from L-tyrosine.In the present study, monophenol monooxygenase from S. lincolnensis has been characterized, and evidence has been presented on the relation of this enzymic activity and the size of tyrosine pool to melanin formation and biosynthesis of lincomycin. It has been found that mutants with lesion in this enzyme do not produce antibiotics.A preliminary account of this work was given at the 2nd International Symposium on Recent I Present address: Fermentation Industry, 02-532 Warsaw, Poland. Progress in Antibiotic Research, Warsaw, 3-5 December 1973. MATERIALS AND METHODSStrains and incubation conditions. The following high lincomycin-producing and nonproducing mutants isolated from the melanin-positive (mel+) and melanin-negative (mell) strains of S. lincolnensis were employed: the original strain mel+NCIB 9413:mel+linco+11 selected from NCIB 9413: mel-linco+426 and mel-linco+434, the two spontaneous lincomycin-producing mutant;9 isolated from mellinco+11:mellinco-2 and mel-linco-l0, the spontaneous nonproducing mutants isolated from mel-linco+433. Under our experimental conditions the yields obtained with the lincomycin-producing strains were: 1,...

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Bioconversion of tyrosine into the propylhygric acid moiety of lincomycin

Cited by 36 publications

References 4 publications

The Novel Transcriptional Regulator LmbU Promotes Lincomycin Biosynthesis through Regulating Expression of Its Target Genes in Streptomyces lincolnensis

The Novel Transcriptional Regulator LmbU Promotes Lincomycin Biosynthesis through Regulating Expression of Its Target Genes in Streptomyces lincolnensis

Biosynthesis, synthesis, and biological activities of pyrrolobenzodiazepines

Monophenol Monooxygenase and Lincomysin Biosynthesis in Streptomyces lincolnensis

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