Inactivation of the
            <i>lys7</i>
            Gene, Encoding Saccharopine Reductase in
            <i>Penicillium chrysogenum</i>
            , Leads to Accumulation of the Secondary Metabolite Precursors Piperideine-6-Carboxylic Acid and Pipecolic Acid from α-Aminoadipic Acid

Naranjo, Leopoldo; Valmaseda, Eva Martín de; Casqueiro, Javier; Ullán, Ricardo V.; Lamas-Maceiras, Mónica; Bañuelos, Óscar; Martı́n, Juan F.

doi:10.1128/aem.70.2.1031-1039.2004

Cited by 20 publications

(9 citation statements)

References 35 publications

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“…Although the targeting efficiency obtained is less than in a DPcku70 strain (Hoff et al, 2009) the observed frequencies of homologous recombination are clearly above values that were observed with the wild-type strain (e.g. Casqueiro et al, 1999;Hoff et al, 2009;Naranjo et al, 2004;Snoek et al, 2009). The use of the non-integrative RNAi vector leads to the construction of single and double mutants that will be a valuable tool for further functional and physiological studies.…”

Section: Discussionmentioning

confidence: 90%

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

2009

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RNA interference (RNAi) is a sequence-specific post-transcriptional gene silencing system that downregulates target gene expression. Here, we provide several lines of evidence for RNA silencing in the industrial β-lactam antibiotic producer Penicillium chrysogenum using the DsRed reporter gene under the control of the constitutive trpC promoter or the inducible xylP promoter. The functional RNAi system was verified by detection of siRNAs that hybridized exclusively with gene-specific 32P-labelled RNA probes. Moreover, when RNAi was used to silence the endogenous PcbrlA morphogene that controls conidiophore development, a dramatic reduction in the formation of conidiospores was observed in 47 % of the corresponding transformants. Evidence that RNAi in P. chrysogenum is dependent on a Dicer peptide was provided with a strain lacking Pcdcl2. In the ΔPcdcl2 background, silencing of the PcbrlA gene was tested. None of the transformants analysed showed a developmental defect. The applicability of the RNAi system in P. chrysogenum was finally demonstrated by silencing the Pcku70 gene to increase homologous recombination frequency. This led to the generation of single and double knockout mutants.

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Section: Discussionmentioning

confidence: 90%

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

2009

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“…3). Later on, this group observed the accumulation of pipecolic acid (along with P6C) upon inactivation of the gene encoding saccharopine reductase, and its origin was determined to be from a-AAA instead of lysine through comparison of the intracellular accumulation of pipecolic acid with another mutant which lacks a-AAA reductase [55]. Here, pipecolic acid appeared to be an incidental intermediate in lysine biosynthesis, derived from the spontaneous chemical equilibrium of a-AAA-semialdehyde into P6C, and the easy conversion of P6C to pipecolic acid (probably catalyzed by the universally existing P5C reductase).…”

Section: A-aminoadipic Acid Pathwaymentioning

confidence: 98%

Pipecolic acid in microbes: biosynthetic routes and enzymes

2006

J IND MICROBIOL BIOTECHNOL

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Pipecolic acid is an important precursor of many useful microbial secondary metabolites. Pipecolic acid-derived moieties are often crucial for the biological activities of some microbial natural products with pharmaceutical applications. Understanding the biogenesis of pipecolic acid in microorganisms would be a significant step toward the mutasynthesis of novel analogs of choice. This review focuses on various microbial pathways and enzymes for pipecolic acid synthesis, especially those related to the origination of pipecolic acid moieties in secondary metabolites.

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“…It has been reported that P. chrysogenum has a very active alpha-aminoadipic acid pathway, which is used for the synthesis of this precursor of penicillin [26]. Saccharopine reductase (SR), an enzyme of the α-aminoadipic acid pathway of lysine biosynthesis in fungus, is involved in lysine degradation pathway to produce aminoadipate, which serves as a key intermediate in the biosynthesis of lysine and penicillin in β-lactam-producing fungi [27].…”

Section: Discussionmentioning

confidence: 99%

Comprehensive Profiling of Proteome Changes Provide Insights of Industrial Penicillium chrysogenum During Pilot and Industrial Penicillin G Fermentation

Cheng

Zhao

Qiao

et al. 2016

Appl Biochem Biotechnol

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The intracellular proteomes of the Penicillium chrysogenum throughout pilot and industrial processes were investigated by using 2-DE combined with MALDI-TOF-TOF MS, respectively. We detected a total of 223 spots corresponding to 154 proteins and 231 spots corresponding to 157 proteins throughout pilot and industrial processes, respectively. The levels of glyceraldehyde-3-phosphate dehydrogenase increased (5.1- and 2.5-fold) under the pilot process, while its levels were no significant changes under the industrial process at 140 and 170 h when compared with that at 2 h. The levels of isocitrate lyase and fumarate hydratase were increased significantly under the industrial process, while their levels had no obvious changes after 20 h of fermentation throughout the pilot process. These results indicate that there were remarkable differences in carbohydrate metabolism (including glycolysis, gluconeogenesis, pentose phosphate pathway, and tricarboxylic acid cycle) of P. chrysogenum during the pilot and industrial fermentations, which likely result in alterations of the primary metabolism and penicillin biosynthesis. Moreover, the differences in the levels of proteins involved in amino acid metabolisms (including valine, cysteine, and α-aminoadipic acid biosynthesis) indicated that the pilot and industrial processes influenced the supplies of penicillin precursors. Compared with that at 2 h, the maximum levels of superoxide (6.9-fold, at 32 h) and catalase (9-fold, at 80 h) during the industrial process and the maximum levels of superoxide (1.2-fold, at 20 h) and catalase (7.7-fold at 128 h) during the pilot process revealed the significant difference in cell redox homeostasis and stress responses during scale-up fermentation. Particularly, 10 spots corresponding to isopenicillin N synthetase and 4 spots corresponding to isopenicillin N (IPN) acyltransferase in pilot and industrial processes were identified, respectively. The levels of IPN acyltransferase (spots 197 and 198) and CoA ligase at 80 h during the industrial process were around 2-fold of that during the pilot process, indicating that the industrial process with a higher penicillin production per cell might provide available environments to induce over-expression of IPN acyltransferase and accelerate penicillin formation. These results provide new insights into the globally potential responses of P. chrysogenum to variations of environments in different fermentation scales so as to consequently regulate the penicillin production.

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Inactivation of the lys7 Gene, Encoding Saccharopine Reductase in Penicillium chrysogenum , Leads to Accumulation of the Secondary Metabolite Precursors Piperideine-6-Carboxylic Acid and Pipecolic Acid from α-Aminoadipic Acid

Cited by 20 publications

References 35 publications

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

Pipecolic acid in microbes: biosynthetic routes and enzymes

Comprehensive Profiling of Proteome Changes Provide Insights of Industrial Penicillium chrysogenum During Pilot and Industrial Penicillin G Fermentation

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