Cytosolic NADPH metabolism in penicillin-G producing and non-producing chemostat cultures of Penicillium chrysogenum

2007 ◽  
Vol 9 (1) ◽  
pp. 112-123 ◽  
Author(s):  
Roelco J. Kleijn ◽  
Feng Liu ◽  
Wouter A. van Winden ◽  
Walter M. van Gulik ◽  
Cor Ras ◽  
...  
Keyword(s):  
Penicillium Chrysogenum ◽  
Penicillin G ◽  
Chemostat Cultures ◽  
Nadph Metabolism

scholarly journals Resolving Phenylalanine Metabolism Sheds Light on Natural Synthesis of Penicillin G in Penicillium chrysogenum

2011 ◽  
Vol 11 (2) ◽  
pp. 238-249 ◽  
Author(s):  
Tânia Veiga ◽  
Daniel Solis-Escalante ◽  
Gabriele Romagnoli ◽  
Angela ten Pierick ◽  
Mark Hanemaaijer ◽  
...  
Keyword(s):  
Nitrogen Source ◽  
Transcriptome Analysis ◽  
Penicillium Chrysogenum ◽  
Penicillin G ◽  
Side Chain ◽  
Acid Decarboxylase ◽  
Content Type ◽  
Chemostat Cultures ◽  
Phenylpyruvate Decarboxylase ◽  
Dual Substrate

ABSTRACTThe industrial production of penicillin G byPenicillium chrysogenumrequires the supplementation of the growth medium with the side chain precursor phenylacetate. The growth ofP. chrysogenumwith phenylalanine as the sole nitrogen source resulted in the extracellular production of phenylacetate and penicillin G. To analyze this natural pathway for penicillin G production, chemostat cultures were switched to [U-13C]phenylalanine as the nitrogen source. The quantification and modeling of the dynamics of labeled metabolites indicated that phenylalanine was (i) incorporated in nascent protein, (ii) transaminated to phenylpyruvate and further converted by oxidation or by decarboxylation, and (iii) hydroxylated to tyrosine and subsequently metabolized via the homogentisate pathway. The involvement of the homogentisate pathway was supported by the comparative transcriptome analysis ofP. chrysogenumcultures grown with phenylalanine and with (NH4)2SO4as the nitrogen source. This transcriptome analysis also enabled the identification of two putative 2-oxo acid decarboxylase genes (Pc13g9300 and Pc18g01490). cDNAs of both genes were cloned and expressed in the 2-oxo-acid-decarboxylase-freeSaccharomyces cerevisiaestrain CEN.PK711-7C (pdc1 pdc5 pdc6Δ aro10Δ thi3Δ). The introduction of Pc13g09300 restored the growth of thisS. cerevisiaemutant on glucose and phenylalanine, thereby demonstrating that Pc13g09300 encodes a dual-substrate pyruvate and phenylpyruvate decarboxylase, which plays a key role in an Ehrlich-type pathway for the production of phenylacetate inP. chrysogenum. These results provide a basis for the metabolic engineering ofP. chrysogenumfor the production of the penicillin G side chain precursor phenylacetate.

scholarly journals Impact of Velvet Complex on Transcriptome and Penicillin G Production in Glucose-Limited Chemostat Cultures of a β-Lactam High-Producing Penicillium chrysogenum Strain

2012 ◽  
Vol 16 (6) ◽  
pp. 320-333 ◽  
Author(s):  
Tânia Veiga ◽  
Jeroen G. Nijland ◽  
Arnold J.M. Driessen ◽  
Roel A.L. Bovenberg ◽  
Hesselein Touw ◽  
...  
Keyword(s):  
Penicillium Chrysogenum ◽  
Penicillin G ◽  
Chemostat Cultures ◽  
Velvet Complex

scholarly journals Kloning Gen pcbC dari Penicillium chrysogenum ke dalam Plasmid pPICZA untuk Pengembangan Produksi Penisilin G

2014 ◽  
Vol 16 (1) ◽  
pp. 33
Author(s):  
Risma Wiharyani ◽  
Dudi Hardianto ◽  
Hermin Pancasakti Kusumaningrum ◽  
Anto Budiharjo
Keyword(s):  
Dna Sequences ◽  
Penicillium Chrysogenum ◽  
Raw Materials ◽  
Raw Material ◽  
Penicillin G ◽  
Gene Fragment ◽  
Semisynthetic Penicillin ◽  
Plasmid Amplification ◽  
Blast Program ◽  
Important Enzyme

Availability of drugs in Indonesia is still limited by the high prices of drugs due to on the imported raw materials that reaches 95%. Developing antibiotic raw materials can be achieved by increasing of penicillin G production, which is the raw material for the formation of semisynthetic penicillin derivatives through the production of 6-aminopenisillanic acid (6-APA). One of the important enzyme in the penicillin G biosynthesis is Isopenisilin N Synthase (IPNS) that encodes by pcbC gene on Penicillium chrysogenum. This study aimed to obtain a recombinant of pcbC gene fragments that is inserted into pPICZA plasmid. Amplification of pcbC gene used pcbC-F and pcbC-R primers. The pcbC gene fragment was inserted into pPICZA vector and then transformed into TOP 10 F’. The results showed that the recombinant of the pcbC gene fragment from P. chrysogenum has been obtained. Analysis of DNA sequences using the BLAST program showed that the pcbC gene fragment has high homology (99%) with the  pcbC gene from P. chrysogenum Wisconsin 54-1255 and P. chrysogenum AS-P-78 which encodes IPNS   Keywords: pcbC Gene, Penicillium chrysogenum, cloning, penicillin G

A Method for Determination of Penicillin G Residue in Waste Penicillium chrysogenum Using High Performance Liquid Chromatography

2015 ◽  
Vol 768 ◽  
pp. 15-24
Author(s):  
Pu Wang ◽  
Hui Ling Liu ◽  
Bing Wang ◽  
Xiu Wen Cheng ◽  
Qing Hua Chen ◽  
...  
Keyword(s):  
Penicillium Chrysogenum ◽  
High Performance ◽  
Extraction Process ◽  
Single Step ◽  
Penicillin G ◽  
Selective Method ◽  
Extraction Solvent ◽  
Oasis Hlb ◽  
Optimized Conditions

In this study, a rapid and selective method has been developed to determine PENG residues in waste penicillium chrysogenum by using SPE cleanup strategy followed by HPLC. Furthermore, some parameters which influenced the extraction efficiency including extraction mode, solvent and time, while washing solution and eluting solution for SPE were systematically investigated. It should be noted that the extraction process was carried out in a single step by mixing the extraction solvent acetonitrile: formic acid in aqueous solution and chrysogenum samples under ultrasound. The SPE procedure was conducted using Oasis HLB as the clean up cartridge, n-hexane as washing solution, and mixture of acetonitrile and methanol as eluting solution. Under the optimized conditions, the linear of PENG are in the range of 0.1-2000 μg/mL, with the correlation was R2>0.99. In addition, the recoveries of PENG in these samples at three fortification levels of 800-1800mg/kg were 74.98% to 113.47% are obtained, respectively. Moreover, a limits of detection (0.006 mg/kg) and quantification (0.02 mg/kg) could be achieved.

scholarly journals Resolving Phenylalanine Metabolism Sheds Light on Natural Synthesis of Penicillin G in Penicillium chrysogenum

2013 ◽  
Vol 12 (1) ◽  
pp. 151-151
Author(s):  
Tânia Veiga ◽  
Daniel Solis-Escalante ◽  
Gabriele Romagnoli ◽  
Angela ten Pierick ◽  
Mark Hanemaaijer ◽  
...  
Keyword(s):  
Penicillium Chrysogenum ◽  
Penicillin G ◽  
Phenylalanine Metabolism

Direct Enzymatic Synthesis of Penicillin G Using Cyclases of Penicillium chrysogenum and Acremonium chrysogenum

1986 ◽  
Vol 4 (1) ◽  
pp. 44-47 ◽  
Author(s):  
J. M. Luengo ◽  
M. T. Alemany ◽  
F. Salto ◽  
F. Ramos ◽  
M. J. López-Nieto ◽  
...  
Keyword(s):  
Enzymatic Synthesis ◽  
Penicillium Chrysogenum ◽  
Penicillin G ◽  
Acremonium Chrysogenum

scholarly journals Synthesis of Antibiotic Penicillin-G Enzymatically by Penicillium chrysogenum

2019 ◽  
Vol 31 (10) ◽  
pp. 2367-2369 ◽  
Author(s):  
Refdinal Nawfa ◽  
Adi Setyo Purnomo ◽  
Herdayanto Sulistyo Putro
Keyword(s):  
Penicillium Chrysogenum ◽  
Research Method ◽  
Biosynthetic Pathway ◽  
Phenylacetic Acid ◽  
Penicillin G ◽  
Antibiotic Concentration ◽  
Food Ingredients ◽  
Enzymatic Formation ◽  
Penicillanic Acid ◽  
Amino Acid Combination

Penicillin-G antibiotic was used as the basic ingredient of making antibiotic type β-lactam such as tetracycline, amoxicillin, ampicillin and other antibiotics. Penicillin-G was splited into 6-amino penicillanic acid as the source of β-lactam. The biosynthetic pathway for the formation of penicillin-G in Penicillium chrysogenum cell through the formation of intermediates was carried out in the form of amino acids such as α-aminoadipate, L-cysteine, L-valine which are formed from glucose (food ingredients).The formation of 6-amino penicillanic acid is an amino acid combination of L-cysteine and L-valine, a step part of the formation of antibiotic penicillin-G in P. chrysogenum cells, thus, it is obvious that there are enzymes involved in its formation. The objective of this study was to examine the use of enzymes present in P. chrysogenum cells to produce penicillin-G and 6-amino penicillanic acid using the intermediate compounds α-aminoadipate, L-cysteine, L-valine and phenylacetic acid assisted by NAFA® coenzymes in P. chrysogenum cells which is more permeable. The research method started from producing biomass of P. chrysogenum cells that demonstrated penicillin-producing antibiotic capability, as the source of the enzyme, followed by addition of permeability treatment of P. chrysogenum cell membrane to get immobile of enzyme by its own cell therefore it can be used more than once. After that the enzyme activity was proven by adding α-aminoadipate, L-cysteine, L-valine, phenylacetic acid and NAFA® coenzyme for the formation of penicillin-G, whereas the addition of L-cystein, L-valine and NAFA® coenzyme were aimed to form 6-amino penicillanic acid. The results showed that P. chrysogenum is able to produce antibiotics with stationary early phase on day 6. The best increased permeability of P. chrysogenum cell membranes was obtained using a 1:4 of toluene:ethanol ratio mixture with the highest antibiotic concentration (130.06 mg/L) after testing for the enzymatic formation of antibacterial penicillin-G.

scholarly journals Formate as an Auxiliary Substrate for Glucose-Limited Cultivation of Penicillium chrysogenum: Impact on Penicillin G Production and Biomass Yield

2007 ◽  
Vol 73 (15) ◽  
pp. 5020-5025 ◽  
Author(s):  
Diana M. Harris ◽  
Zita A. van der Krogt ◽  
Walter M. van Gulik ◽  
Johannes P. van Dijken ◽  
Jack T. Pronk
Keyword(s):  
Penicillium Chrysogenum ◽  
Product Yield ◽  
Penicillin G ◽  
Carbon Substrate ◽  
Molar Ratios ◽  
Formate Oxidation ◽  
System A ◽  
Glucose Ratio ◽  
Biomass Yields ◽  
Substrate Feeding

ABSTRACT Production of β-lactams by the filamentous fungus Penicillium chrysogenum requires a substantial input of ATP. During glucose-limited growth, this ATP is derived from glucose dissimilation, which reduces the product yield on glucose. The present study has investigated whether penicillin G yields on glucose can be enhanced by cofeeding of an auxiliary substrate that acts as an energy source but not as a carbon substrate. As a model system, a high-producing industrial strain of P. chrysogenum was grown in chemostat cultures on mixed substrates containing different molar ratios of formate and glucose. Up to a formate-to-glucose ratio of 4.5 mol·mol−1, an increasing rate of formate oxidation via a cytosolic NAD+-dependent formate dehydrogenase increasingly replaced the dissimilatory flow of glucose. This resulted in increased biomass yields on glucose. Since at these formate-to-glucose ratios the specific penicillin G production rate remained constant, the volumetric productivity increased. Metabolic modeling studies indicated that formate transport in P. chrysogenum does not require an input of free energy. At formate-to-glucose ratios above 4.5 mol·mol−1, the residual formate concentrations in the cultures increased, probably due to kinetic constraints in the formate-oxidizing system. The accumulation of formate coincided with a loss of the coupling between formate oxidation and the production of biomass and penicillin G. These results demonstrate that, in principle, mixed-substrate feeding can be used to increase the yield on a carbon source of assimilatory products such as β-lactams.

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