Effects of anoxic conditions on the enzymatic conversion of D,L-2-amino-thiazoline-4-carboxylic acid to L-cystine

K. H. Nam, O. H. Ryu, J. Park, C. S. Shin

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2 Citations (Scopus)

Abstract

The effects of anoxic conditions on product inhibition and the stability of L-ATC hydrolase were investigated in the conversion of D,L-2-amino-Δ2-thiazoline-4-carboxylic acid (D,L-ATC) to L-cystine using the cell free extract enzyme of Pseudomonas sp. in the presence of hydroxylamine. At L-cysteine equivalent levels, where one mole of L-cystine was counted as two moles of L-cysteine, L-cystine inhibited the L-ATC hydrolase reaction to a greater extent than L-cysteine. In air, the product occurred predominantly as L-cystine (94.9%), whereas in a nitrogen atmosphere the product occurred as a mixture of L-cysteine (39.3%) and L-cystine (40.7%). As a result, less product inhibition took place in nitrogen. The activity of L-ATC hydrolase was almost fully lost after 20 h of incubation by shaking at 30°C in air, but considerable activity remained under the anoxic conditions of nitrogen. A kinetic analysis of the reactions confirmed that reduced product inhibition and enhanced enzyme stability in nitrogen result in a more efficient enzyme reaction. The inactivation rate constant (k1) was estimated to be 0.11 h-1 in nitrogen and 0.22-1 in air, indicating that the stability of L-ATC hydrolase in nitrogen was greater than in air. The values of the K(p1), and K(p2) constants related to product inhibition were 43.36 mM and 30.48 mM for L-cysteine and L-cystine, respectively, where higher values were an indication of less product inhibition. The value of the rate constant (k2) for the oxidation of L-cysteine to L-cystine was 0.09 h-1 in nitrogen and 1.01 h-1 in air, suggesting that the oxidation of L-cysteine to L-cystine proceeds faster in air than in nitrogen.

Original languageEnglish
Pages (from-to)185-193
Number of pages9
JournalActa Biotechnologica
Volume17
Issue number2
DOIs
Publication statusPublished - 1997

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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