Skip to main content
Fachgebiete
Automobil + Motoren Bauwesen + Immobilien Business IT + Informatik Elektrotechnik + Elektronik Energie + Nachhaltigkeit Finance + Banking Management + Führung Marketing + Vertrieb Maschinenbau + Werkstoffe Versicherung + Risiko
DE
EN
Bücher
Zeitschriften
Themenseiten
Organisationspsychologie Projektmanagement Marketing Smart Manufacturing
Weitere Formate
Podcasts Webinare Technik Webinare Wirtschaft Kongresse Veranstaltungskalender Awards
Jetzt Einzelzugang starten
Zugang für Unternehmen
Referenzkunden
SLX-Digitalkonferenz: Zukunftswerkstatt 2024

Mehr Umsatz mit Top-Kontakten

Am 7. Mai erfahren Sie, wie Vertriebsteams Leadgenerierung, Leadnurturing und Leadstrategien effizient steuern können.
Erweiterte Suche
Anmelden
nach oben
Erschienen in:
Present Global Situation of Amino Acids in Industry Kapitel lesen Erstes Kapitel lesen

2017 | OriginalPaper | Buchkapitel

Boosting Anaplerotic Reactions by Pyruvate Kinase Gene Deletion and Phosphoenolpyruvate Carboxylase Desensitization for Glutamic Acid and Lysine Production in Corynebacterium glutamicum

verfasst von : Atsushi Yokota, Kazunori Sawada, Masaru Wada

Erschienen in: Amino Acid Fermentation

Verlag: Springer Japan

Einloggen

Aktivieren Sie unsere intelligente Suche, um passende Fachinhalte oder Patente zu finden.

search-config
loading …

Abstract

In the 1980s, Shiio and coworkers demonstrated using random mutagenesis that the following three phenotypes were effective for boosting lysine production by Corynebacterium glutamicum: (1) low-activity-level citrate synthase (CSL), (2) phosphoenolpyruvate carboxylase (PEPC) resistant to feedback inhibition by aspartic acid (PEPCR), and (3) pyruvate kinase (PYK) deficiency. Here, we reevaluated these phenotypes and their interrelationship in lysine production using recombinant DNA techniques.
The pyk deletion and PEPCR (D299N in ppc) independently showed marginal effects on lysine production, but both phenotypes synergistically increased lysine yield, demonstrating the importance of PEPC as an anaplerotic enzyme in lysine production. Similar effects were also found for glutamic acid production. CSL (S252C in gltA) further increased lysine yield. Thus, using molecular techniques, the combination of these three phenotypes was reconfirmed to be effective for lysine production. However, a simple CSL mutant showed instabilities in growth and lysine yield.
Surprisingly, the pyk deletion was found to increase biomass production in wild-type C. glutamicum ATCC13032 under biotin-sufficient conditions. The mutant showed a 37% increase in growth (based on OD660) compared with the ATCC13032 strain in a complex medium containing 100 g/L glucose. Metabolome analysis revealed the intracellular accumulation of excess precursor metabolites. Thus, their conversion into biomass was considered to relieve the metabolic distortion in the pyk-deleted mutant. Detailed physiological studies of various pyk-deleted mutants also suggested that malate:quinone oxidoreductase (MQO) is important to control both the intracellular oxaloacetic acid (OAA) level and respiration rate. These findings may facilitate the rational use of C. glutamicum in fermentation industries.

Sie haben noch keine Lizenz? Dann Informieren Sie sich jetzt über unsere Produkte:

Springer Professional "Wirtschaft+Technik"

Online-Abonnement

Mit Springer Professional "Wirtschaft+Technik" erhalten Sie Zugriff auf:

  • über 102.000 Bücher
  • über 537 Zeitschriften

aus folgenden Fachgebieten:

  • Automobil + Motoren
  • Bauwesen + Immobilien
  • Business IT + Informatik
  • Elektrotechnik + Elektronik
  • Energie + Nachhaltigkeit
  • Finance + Banking
  • Management + Führung
  • Marketing + Vertrieb
  • Maschinenbau + Werkstoffe
  • Versicherung + Risiko

Jetzt Wissensvorsprung sichern!

Jetzt informieren

Springer Professional "Technik"

Online-Abonnement

Mit Springer Professional "Technik" erhalten Sie Zugriff auf:

  • über 67.000 Bücher
  • über 390 Zeitschriften

aus folgenden Fachgebieten:

  • Automobil + Motoren
  • Bauwesen + Immobilien
  • Business IT + Informatik
  • Elektrotechnik + Elektronik
  • Energie + Nachhaltigkeit
  • Maschinenbau + Werkstoffe




 

Jetzt Wissensvorsprung sichern!

Jetzt informieren
Vorheriges Kapitel l-Methionine Production
Nächstes Kapitel Exporters for Production of Amino Acids and Other Small Molecules
Literatur
1.
Ozaki H, Shiio I (1983) Production of lysine by pyruvate kinase mutants of Brevibacterium flavum. Agric Biol Chem 47:1569–1576
2.
Shiio I, Yokota A, Sugimoto S (1987) Effect of pyruvate kinase deficiency on L-lysine productivities of mutants with feedback-resistant aspartokinases. Agric Biol Chem 51:2485–2493
3.
Peters-Wendisch PG, Wendisch VF, Paul S et al (1997) Pyruvate carboxylase as an anaplerotic enzyme in Corynebacterium glutamicum. Microbiology 143:1095–1103CrossRef
4.
Gubler M, Park SM, Jetten M et al (1994) Effects of phosphoenol pyruvate carboxylase deficiency on metabolism and lysine production in Corynebacterium glutamicum. Appl Microbiol Biotechnol 40:857–863CrossRef
5.
Peters-Wendisch PG, Schiel B, Wendisch VF et al (2001) Pyruvate carboxylase is a major bottleneck for glutamate and lysine production by Corynebacterium glutamicum. J Mol Microbiol Biotechnol 3:295–300PubMed
6.
Shiio I, Yoshino H, Sugimoto S (1990) Isolation and properties of lysine-producing mutants with feedback-resistant aspartokinase derived from a Brevibacterium flavum strain with citrate synthase- and pyruvate kinase-defects and feedback-resistant phosphoenolpyruvate carboxylase. Agric Biol Chem 54:3275–3282CrossRef
7.
Gubler M, Jetten M, Lee SH et al (1994) Cloning of the pyruvate kinase gene (pyk) of Corynebacterium glutamicum and site-specific inactivation of pyk in a lysine-producing Corynebacterium lactofermentum strain. Appl Environ Microbiol 60:2494–2500PubMedPubMedCentral
8.
Park SM, Sinskey AJ, Stephanopoulos G (1997) Metabolic and physiological studies of Corynebacterium glutamicum mutants. Biotechnol Bioeng 55:864–879CrossRefPubMed
9.
Becker J, Klopprogge C, Wittmann C (2008) Metabolic responses to pyruvate kinase deletion in lysine producing Corynebacterium glutamicum. Microb Cell Fact 7:8CrossRefPubMedPubMedCentral
10.
Ohnishi J, Mitsuhashi S, Hayashi M et al (2002) A novel methodology employing Corynebacterium glutamicum genome information to generate a new L-lysine-producing mutant. Appl Microbiol Biotechnol 58:217–223CrossRefPubMed
11.
Chen Z, Bommareddy RR, Frank D et al (2014) Deregulation of feedback inhibition of phosphoenolpyruvate carboxylase for improved lysine production in Corynebacterium glutamicum. Appl Environ Microbiol 80:1388–1393CrossRefPubMedPubMedCentral
12.
Ooyen J, Noack S, Bott M et al (2012) Improved L-lysine production with Corynebacterium glutamicum and systemic insight into citrate synthase flux and activity. Biotechnol Bioeng 109:2070–2081CrossRefPubMed
13.
Yokota A, Shiio I (1988) Effects of reduced citrate synthase activity and feedback-resistant phosphoenolpyruvate carboxylase on lysine productivities of Brevibacterium flavum mutants. Agric Biol Chem 52:455–463
14.
Sawada K, Zen-in S, Wada M et al (2010) Metabolic changes in a pyruvate kinase gene deletion mutant of Corynebacterium glutamicum ATCC 13032. Metab Eng 12:401–407CrossRefPubMed
15.
Mori M, Shiio I (1984) Production of aspartic acid and enzymatic alteration in pyruvate kinase mutants of Brevibacterium flavum. Agric Biol Chem 48:1189–1197
16.
Wada M, Sawada K, Ogura K et al (2016) Effects of phosphoenolpyruvate carboxylase desensitization on glutamic acid production in Corynebacterium glutamicum ATCC 13032. J Biosci Bioeng 121:172–177CrossRefPubMed
17.
Yanase M, Aikoh T, Sawada K et al (2016) Pyruvate kinase deletion as an effective phenotype to enhance lysine production in Corynebacterium glutamicum ATCC13032: redirecting the carbon flow to a precursor metabolite. J Biosci Bioeng 122:160–167CrossRefPubMed
18.
Shiio I, Ozaki H, Ujigawa-Takeda K (1982) Production of aspartic acid and lysine by citrate synthase mutants of Brevibacterium flavum. Agric Biol Chem 46:101–107
19.
Radmacher E, Eggeling L (2007) The three tricarboxylate synthase activities of Corynebacterium glutamicum and increase of L-lysine synthesis. Appl Microbiol Biotechnol 76:587–595CrossRefPubMed
20.
Sawada K, Wada M, Hagiwara T et al (2015) Effect of pyruvate kinase gene deletion on the physiology of Corynebacterium glutamicum ATCC13032 under biotin-sufficient non-glutamate-producing conditions: enhanced biomass production. Metab Eng Commun 2:67–75CrossRef
21.
Molenaar D, van der Rest ME, Drysch A et al (2000) Functions of the membrane-associated and cytoplasmic malate dehydrogenases in the citric acid cycle of Corynebacterium glutamicum. J Bacteriol 182:6884–6891CrossRefPubMedPubMedCentral
22.
Sawada K, Kato Y, Imai K et al (2012) Mechanism of increased respiration in an H+-ATPase-defective mutant of Corynebacterium glutamicum. J Biosci Bioeng 113:467–473CrossRefPubMed
Metadaten
Titel
Boosting Anaplerotic Reactions by Pyruvate Kinase Gene Deletion and Phosphoenolpyruvate Carboxylase Desensitization for Glutamic Acid and Lysine Production in Corynebacterium glutamicum
verfasst von
Atsushi Yokota
Kazunori Sawada
Masaru Wada
Copyright-Jahr
2017
Verlag
Springer Japan
Buch
Amino Acid Fermentation

Print ISBN: 978-4-431-56518-5

Electronic ISBN: 978-4-431-56520-8

Copyright-Jahr: 2017

DOI
https://doi.org/10.1007/10_2016_31

Premium Partner

    Marktübersichten

    Die im Laufe eines Jahres in der „adhäsion“ veröffentlichten Marktübersichten helfen Anwendern verschiedenster Branchen, sich einen gezielten Überblick über Lieferantenangebote zu verschaffen. 

    Zur Marktübersicht
    Bildnachweise