Regulation of coenzyme A biosynthesis

Jackowski, Suzanne; Rock, Charles O.

doi:10.1128/jb.148.3.926-932.1981

Cited by 234 publications

(154 citation statements)

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“…As a rate-limiting enzyme, PANK plays an important role in the biosynthesis of CoA and ACP. 34 The CoA-bound PA is involved in the release of energy from carbohydrates, fatty acids and amino acids, and ACP-bound PA is related to the synthesis of fatty acids. 2 Therefore, the linear increase in PANK activity was consistent with the improved acetyl CoA, succinyl CoA and ACP, and suggested that supplementary RPP improved the utilisation of energy and nutrient.…”

Section: Discussionmentioning

confidence: 99%

Effects of rumen‐protected pantothenate supplementation on lactation performance, ruminal fermentation, nutrient digestion and blood metabolites in dairy cows

Wang

Liu

et al. 2017

J Sci Food Agric

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Section: Discussionmentioning

confidence: 99%

Effects of rumen‐protected pantothenate supplementation on lactation performance, ruminal fermentation, nutrient digestion and blood metabolites in dairy cows

Wang

Liu

et al. 2017

J Sci Food Agric

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“…E. coli RV308 AlucX74gul [18] was obtained from Dr E. D. Laue (Department of Biochemistry, University of Cambridge). E. coli SJ16 panD2 zad-220: :TnlO [19] was obtained from Dr J. E. Cronan Jr (Department of Microbiology, University of Illinois). E. coli strains were generally grown on 1% yeast extract, 1% tryptone and 0.5% NaCl either at 37°C or 30°C.…”

Section: Methodsmentioning

confidence: 99%

Heterologous expression in Escherichia coli of an intact multienzyme component of the erythromycin‐producing polyketide synthase

Roberts

Staunton

Leadlay

1993

European Journal of Biochemistry

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-EJB 92 1720/2 6-Deoxyerythronolide B synthase 3 (DEBS 3) is proposed to catalyse the fifth and sixth condensation cycles in the assembly of the polyketide 6-deoxyerythronolide B, the first isolatable intermediate in the biosynthesis of erythromycin A by Saccharopolyspora erythraea. The gene encoding DEBS 3 has previously been cloned and sequenced, and the deduced product is predicted to house nine fatty acid synthase-like activities on a 330-kDa polypeptide chain. The gene has been engineered into a pT-7-based expression system for over-expression in Escherichia coli. Recombinant DEBS 3 was found to constitute, after induction, 1-2% of soluble intracellular protein. DEBS 3 was purified from extracts of the recombinant E. coli to apparent homogeneity, and was found not to be modified by covalent attachment of the prosthetic group 4'-phosphopantetheine. Incubation with (R,S)-methylmalonyl-CoA, the presumed source of extension units for polyketide chain assembly, led to hydrolysis of the thioester, implying that the methylmalonyl-CoA :ACP acyltransferase domains in DEBS 3 are correctly folded and able to catalyse this side-reaction. During this reaction, DEBS 3 became transiently radiolabelled, consistent with the intermediacy of an acylenzyme. The native molecular mass of the protein by gel filtration chromatography was 668 kDa which corresponds either to a dimer or to a highly asymmetric monomer.Erythromycin A is a clinically important macrolide antibiotic produced by the Gram-positive soil-borne bacterium, Saccharopolyspora erythraea [l]. The biosynthetic pathway to erythromycin A begins with the processive assembly of a polyketide chain from one molecule of propionyl-CoA and six molecules of methylmalonyl-CoA by a mechanism directly analogous to that of fatty acid biosynthesis [2]. Following six rounds of condensation and reduction the polyketide chain is cyclised to give the 14-membered macrolactone, 6-deoxyerythronolide B, the earliest enzyme-free intermediate [3] so far identified. The lactone is subsequently modified by specific hydroxylases, glycosyltransferases and a methyltransferase to give erythromycin A [4].The erythromycin biosynthetic genes are clustered on the chromosome together with the resistance determinant ermE. The genes encoding 6-deoxyerythronolide B synthase (DEBS) are located within the eryA region of the biosynthetic gene cluster and together span over 30 kbp [ 5 ] , representing more than 50% of the total size of the cluster. Partial nucleotide sequencing of this region revealed multiple copies of sequences encoding fatty acid synthase-like activi-

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“…Each of the expression constructs was transformed into E. coli E103S (from Dr Lee Simon, Waksman Institute of Microbiology, Piscataway, New Jersey) and a p-alanine auxotroph, SJ16 [20].…”

Section: Site-spec Ific Mu T Ugmr>sismentioning

confidence: 99%

“…Cultures (3 ml) of E. coli strain SJ16 transformed with pPBl04, pCY104 or pTH104 were grown for 12 h in the presence of 10 pCi [3H]fl-alanine. The cells were harvested and lysed as previously described [20] and the lysate was treated with 100 mM dithiothreitol at pH 9.0. The proteins were then analyzed by SDS/PAGE, the gel treated with En-3Hance (New England Nuclear), dried and exposed to Kodak XAR-5 film.…”

Section: /3-alunine Labellingmentioning

confidence: 99%

“…In order to provide a more sensitive and definitive test for possible phosphopantetheine attachment to these mutants ACPs, SJ16 cells were transformed with pCY104, pTH104, or pPB104. SJ16 is a p-alanine auxotroph and can be used to specifically label the phosphopantetheine prosthetic group of CoA and ACP [20]. Proteins isolated from these SJ16 cells labelled with fl-[3H]alanine were separated on SDS gels.…”

Section: Assuyfor Phosphopantetheine In Mutant Acpsmentioning

confidence: 99%

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Site‐directed mutagenesis of the spinach acyl carrier protein‐I prosthetic group attachment site

Jaworski

Post-Beittenmiller

Ohlrogge

1989

European Journal of Biochemistry

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Site-directed mutagenesis was used to change the phosphopantetheine attachment site (Ser38) of spinach acyl carrier protein I (ACP-I) from a serine to a threonine or cysteine residue.1. Although the native ACP-I is fully phosphopantethenylated when expressed in Escherichia coli, the TH-ACP-I and CY-ACP-I mutants were found to be completely devoid of the phosphopantetheine group. Therefore, the E. coli holoACP synthase requires serine for in vivo phosphopantetheine addition to spinach ACP-I.2. Spinach holoACP synthase was completely inactive in vitro with either the TH-ACP-I or CY-ACP-I mutants. In addition, TH-ACP-I and CY-ACP-I were strong inhibitors of spinach holoACP synthase.3. The mutant ACPs were weak or ineffective as inhibitors of spinach fatty acid synthesis and spinach oleoyl-ACP hydrolase.4. Compared to holoACP-I, the mutant apoACP-I analogs had: (a) altered mobility in SDS and native gel electrophoresis, (b) altered binding to anti-(spinach ACP-I) antibodies and (c) altered isoelectric points. The combined physical, immunological and enzyme inhibition data indicate that attachment of the phosphopantheine prosthetic group alters ACP conformation.Plant acyl carrier protein (ACP) is a small (z 9 kDa), acidic chloroplast protein which serves as a cofactor in at least a dozen reactions of fatty acid synthesis, desaturation and glycerolipid assembly [l]. ACP is the best characterized protein in plant lipid biosynthesis, and cDNA and/or genomic clones are available from spinach [2, 31, barley [4], Brassica [5,6] and Arabidopsis [7]. Multiple isoforms of ACP have been found in plants, and in spinach leaf the most abundant form is designated ACP-I. ACPs contain a phosphopantetheine prosthetic group attached to a serine residue near the middle of the polypeptide chain. This prosthetic group is donated from CoA in a reaction catalyzed by holoACP synthase [S, 91. In plants, ACP is nuclear encoded and is synthesized as a precursor containing a transit peptide for plastid uptake [3 -71. The prosthetic group attachment appears to occur outside the chloroplast before uptake and processing to mature holoACP [8].In E. coli, the enzyme holoACP hydrolase acts to remove the prosthetic group from ACP. However, Jackowski and Rock have demonstrated that there are no detectable levels of apoACP in E. coli cells [lo]. Similarly, we have been unable to detect apoACP in spinach tissues using Western blots probed with antibodies which recognize both holoACP and apoACP (Battey & Ohlrogge, unpublished data). Thus, in both plants and bacteria the activity of holoACP synthase appears sufficient to maintain ACP in a fully pantethenylated form. The E. coli mutant, MP4, which has reduced holoACP

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Regulation of coenzyme A biosynthesis

Cited by 234 publications

References 14 publications

Effects of rumen‐protected pantothenate supplementation on lactation performance, ruminal fermentation, nutrient digestion and blood metabolites in dairy cows

Effects of rumen‐protected pantothenate supplementation on lactation performance, ruminal fermentation, nutrient digestion and blood metabolites in dairy cows

Heterologous expression in Escherichia coli of an intact multienzyme component of the erythromycin‐producing polyketide synthase

Site‐directed mutagenesis of the spinach acyl carrier protein‐I prosthetic group attachment site

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