Fermentative Production of Tryptophan by a Stable Recombinant Strain of Corynebacterium glutamicumwith a Modified Serine-biosynthetic Pathway

Masato Ikeda, Kuniki Kino, Keiko Nakanishi, Ryoichi Katsumata

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Introduction of plasmid pKW99, which coexpresses the deregulated 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase and tryptophan-biosynthetic enzymes, into tryptophan-producing Corynebacterium glutamicumKY10894 resulted in a marked increase (54%) in yield of tryptophan production (43 g/liter), but incurred two problems. One was a decline in sugar consumption at the late stage of fermentation, and the other the loss of the plasmid in the absence of selective pressure. The retarded sugar assimilation was found to be attributed to the death of cells that arose from the detrimental action of indole, the last intermediate in the tryptophan pathway, accumulated as a by-product. A chain of these events simultaneously disappeared when serine, the other substrate of the final reaction by tryptophan synthase, was added. These results indicated that a limiting supply of serine was the cause of the decline in the sugar consumption. Thus, to increase carbon flux into serine, the gene for 3-phosphoglycerate dehydrogenase (PGD), the first enzyme in the serine pathway, was cloned from wild-type C. glutamicumATCC 31833 and joined onto pKW99 to generate pKW9901. Strain KY10894 transformed with pKW9901 favorably consumed sugar through fermentation with accumulating little indole. Furthermore, on the basis of the observation that serine in the medium was consumed rapidly by the recombinant cells, we developed a unique plasmid stabilization system composed of KY9218 (a PGD-deficient serine-requiring strain of KY10894) and pKW9901: In its combination, cells lacking the plasmid should not proliferate in the fermentation medium which does not contain serine. Even if selective pressure was not applied, the modified strain KY9218 with pKW9901 stably maintained the plasmid during fermentation and produced 50 g/liter of tryptophan in a 61% increased yield relative to strain KY10894.

Original languageEnglish
Pages (from-to)674-678
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Volume58
Issue number4
DOIs
Publication statusPublished - 1994
Externally publishedYes

Fingerprint

Corynebacterium
Biosynthetic Pathways
Tryptophan
Serine
Plasmids
Sugars
Fermentation
Phosphoglycerate Dehydrogenase
3-Deoxy-7-Phosphoheptulonate Synthase
Tryptophan Synthase
Carbon Cycle
Enzymes
Byproducts
Cell Death
Carbon
Stabilization
Genes
Cells
Fluxes
Substrates

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Fermentative Production of Tryptophan by a Stable Recombinant Strain of Corynebacterium glutamicumwith a Modified Serine-biosynthetic Pathway. / Ikeda, Masato; Kino, Kuniki; Nakanishi, Keiko; Katsumata, Ryoichi.

In: Bioscience, Biotechnology and Biochemistry, Vol. 58, No. 4, 1994, p. 674-678.

Research output: Contribution to journalArticle

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