Biochemical characteristization of propionyl-coenzyme a carboxylase complex of Streptomyces toxytricini

Biotin-dependent carboxylases include acetyl-CoA carboxylase (ACC), propionyl-CoA carboxylase (PCC), 3-methylcrotonyl-CoA carboxylase (MCC), geranyl-CoA carboxylase (GCC), pyruvate carboxylase (PC), and urea carboxylase (UC). They contain biotin carboxylase (BC), carboxyltransferase (CT) and biotin-carboxyl carrier protein (BCCP) components. These enzymes are widely distributed in nature and have important functions in fatty acid metabolism, amino acid metabolism, carbohydrate metabolism, polyketide biosynthesis, urea utilization, and other cellular processes. ACCs are also attractive targets for drug discovery against type 2 diabetes, obesity, cancer, microbial infections, and other diseases, and the plastid ACC of grasses is the target of action of three classes of commercial herbicides. Deficiencies in the activities of PCC, MCC or PC are linked to serious diseases in humans. Our understanding of these enzymes has been greatly enhanced over the past few years by the crystal structures of the holoenzymes of PCC, MCC, PC, and UC. The structures reveal unanticipated features in the architectures of the holoenzymes, including the presence of previously unrecognized domains, and provide a molecular basis for understanding their catalytic mechanism as well as the large collection of disease-causing mutations in PCC, MCC and PC. This review will summarize the recent advances in our knowledge on the structure and function of these important metabolic enzymes.

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“…2) [12,229]. These enzymes are different from PCC (family 1.5) in that they probably lack the BT domain in the α subunit.…”

Section: Other Acyl-coa Carboxylasesmentioning

confidence: 99%

Structure and function of biotin-dependent carboxylases

Tong

2012

Cell. Mol. Life Sci.

373

338

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“…Although the removal of GlnB protein relieved its inhibition of ACC and PCC, increasing the yield of FK520 to a certain extent, the content of methylmalonyl-CoA in the late period of fermentation was still low ( Figure 4D ). Previous research suggested that the specific β subunit (responsible for substrate recognition) and ε subunit (responsible for protein- protein interactions) are crucial for the carboxylation activity of PCC ( Diacovich et al, 2002 ; Demirev et al, 2011 ; Tong, 2013 ). The results of amino acid sequence alignment showed that the 1_3398 gene ( pccB ) encoded the β subunit, while the 1_3397 gene ( pccE ) encoded the ε subunit of PCC in S. hygroscopicus var.…”

Section: Resultsmentioning

confidence: 99%

“…Accordingly, it was speculated that ACC and PCC might both be regulated by PII proteins in actinobacteria ( Figure 1A ). Since the carboxylation products of these two enzymes, malonyl-CoA and methylmalonyl-CoA, are important precursors for the biosynthesis of polyketide antibiotics ( Rodriguez et al, 2001 ; Demirev et al, 2011 ), it is of great significance to explore the influence of PII proteins on the activity of ACC and PCC, with the aim of improving the antibiotic yield.…”

Section: Introductionmentioning

confidence: 99%

Increasing the Ascomycin Yield by Relieving the Inhibition of Acetyl/Propionyl-CoA Carboxylase by the Signal Transduction Protein GlnB

Wang

Yin

et al. 2021

Front. Microbiol.

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Ascomycin (FK520) is a multifunctional antibiotic produced by Streptomyces hygroscopicus var. ascomyceticus. In this study, we demonstrated that the inactivation of GlnB, a signal transduction protein belonging to the PII family, can increase the production of ascomycin by strengthening the supply of the precursors malonyl-CoA and methylmalonyl-CoA, which are produced by acetyl-CoA carboxylase and propionyl-CoA carboxylase, respectively. Bioinformatics analysis showed that Streptomyces hygroscopicus var. ascomyceticus contains two PII family signal transduction proteins, GlnB and GlnK. Protein co-precipitation experiments demonstrated that GlnB protein could bind to the α subunit of acetyl-CoA carboxylase, and this binding could be disassociated by a sufficient concentration of 2-oxoglutarate. Coupled enzyme activity assays further revealed that the interaction between GlnB protein and the α subunit inhibited both the activity of acetyl-CoA carboxylase and propionyl-CoA carboxylase, and this inhibition could be relieved by 2-oxoglutarate in a concentration-dependent manner. Because GlnK protein can act redundantly to maintain metabolic homeostasis under the control of the global nitrogen regulator GlnR, the deletion of GlnB protein enhanced the supply of malonyl-CoA and methylmalonyl-CoA by restoring the activity of acetyl-CoA carboxylase and propionyl-CoA carboxylase, thereby improving the production of ascomycin to 390 ± 10 mg/L. On this basis, the co-overexpression of the β and ε subunits of propionyl-CoA carboxylase further increased the ascomycin yield to 550 ± 20 mg/L, which was 1.9-fold higher than that of the parent strain FS35 (287 ± 9 mg/L). Taken together, this study provides a novel strategy to increase the production of ascomycin, providing a reference for improving the yield of other antibiotics.

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“…Such proteins act as carboxyl transferases involved in precursor supply in fatty acid or polyketide biosynthesis. [12] Phn2 revealed 47 % identity to a gene encoding for a PKS of Streptomyces violaceusniger Tu 4113 (Table 1). A detailed analysis of the 22 kb gene gave evidence for the presence of a complete PKS consisting of five modules.…”

Section: Sequence Analysis Of the Phenylnannolone A Biosynthetic Genesmentioning

confidence: 99%

Biosynthesis of Phenylnannolone A, a Multidrug Resistance Reversal Agent from the Halotolerant Myxobacterium Nannocystis pusilla B150

et al. 2014

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The myxobacterial strain Nannocystis pusilla B150 synthesizes the structurally new polyketides phenylnannolone A–C. Apart from some common volatiles and siderophores, these are the first natural products from the genus Nannocystis. Phenylnannolone A shows inhibitory activity towards the ABCB1 gene product P‐glycoprotein and reverses daunorubicin resistance in cancer cells. To decipher the biochemical reactions leading to the formation of phenylnannolone A, the putative biosynthetic genes were identified (phn1, phn2). Phn2 is a polyketide synthase (PKS) with an NRPS‐like loading module, and its domain order is consistent with the phenylnannolone A structure. The functionality and substrate selectivity of the loading module were determined by means of a γ‐18O4‐ATP pyrophosphate exchange and a phosphopantetheine ejection assay. A specific activation of cinnamic acid by the AMP‐ligase was detected. Phn1 is a putative butyryl‐CoA carboxylase (BCC), providing ethylmalonyl‐CoA for the formation of the ethyl‐substituted part of phenylnannolone A. Phn1 is the first BCC found in biosynthetic genes for an ethyl‐substituted natural compound. Biosynthesis of phenylnannolone A, putatively encoded by phn1 and phn2, thus utilizes the first biosynthetic machinery in which both a BCC and a PKS are involved.

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Biochemical characteristization of propionyl-coenzyme a carboxylase complex of Streptomyces toxytricini

Cited by 11 publications

References 19 publications

Structure and function of biotin-dependent carboxylases

Structure and function of biotin-dependent carboxylases

Increasing the Ascomycin Yield by Relieving the Inhibition of Acetyl/Propionyl-CoA Carboxylase by the Signal Transduction Protein GlnB

Biosynthesis of Phenylnannolone A, a Multidrug Resistance Reversal Agent from the Halotolerant Myxobacterium Nannocystis pusilla B150

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