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Literature summary for 1.7.3.3 extracted from

  • Chen, Z.; Wang, Z.; He, X.; Guo, X.; Li, W.; Zhang, B.
    Uricase production by a recombinant Hansenula polymorpha strain harboring Candida utilis uricase gene (2008), Appl. Microbiol. Biotechnol., 79, 545-554.
    View publication on PubMed

Application

Application Comment Organism
medicine uricase is an important medical enzyme which can be used to determine urate in clinical analysis, to therapy gout, hyperuricemia, and tumor lysis syndrome Cyberlindnera jadinii

Cloned(Commentary)

Cloned (Comment) Organism
uricase production by the recombinant Hansenula polymorpha strain MU200 harboring Candida utilis uricase gene under the control of methanol oxidase promoter using Saccharomyces cerevisiae alpha-factor signal peptide as the secretory sequence Cyberlindnera jadinii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34000
-
recombinant uricase determined by SDS-PAGE Cyberlindnera jadinii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Cyberlindnera jadinii enzyme catalyzes the oxidation of uric acid to a more soluble and easily excreted compound, allantoin ?
-
?
uric acid + O2 + H2O Cyberlindnera jadinii purine degradation 5-hydroxyisourate + H2O2
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?

Organism

Organism UniProt Comment Textmining
Cyberlindnera jadinii
-
-
-

Source Tissue

Source Tissue Comment Organism Textmining
cell culture strain CGMCC 2.120 is used as source of uricase gene Cyberlindnera jadinii
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
among the parameters investigated in shaking flask cultures, the pH value of medium and inoculum size has great influence on the recombinant uricase production, the maximum extracellular uricase yield of 2.6 U/ml is obtained in shaking flask culture Cyberlindnera jadinii
additional information
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at pH 5.5, the extracellular uricase production reaches top of 7.5 U/ml at 58 h, when fermentation is performed at pH 6.5 for 62 h, 14.5 U/ml of extracellular uricase and 23.3 U/ml of intracellular uricase are produced, the total specific uricase production at pH 6.5 is 1.7times of that at pH 5.5 Cyberlindnera jadinii
additional information
-
in high density fermentation in YPG medium at 37°C, extracellular uricase activity increases significantly during the first 40 h, highest extracellular uricase level of 52.3 U/ml is obtained after 58 h of induction, as well as the intracellular activity of 60.3 U/ml, after 86 h of fermentation and 58 h of induction, a total uricase activity of 112600 U/l is obtained, the extracellular and intracellular yields of uricase in high cell density fermentation increased by 3.7fold and 3.5fold compared with the batch fermentation Cyberlindnera jadinii
additional information
-
the combined use of fed-batch culture and pH-controlled strategy increases the expression level of uricase significantly, the extracellular uricase production of 52.3 U/ml (approximately 2.1 g/l of protein) is obtained, which is much higher than that produced by recombinant Escherichia coli strains Cyberlindnera jadinii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information enzyme catalyzes the oxidation of uric acid to a more soluble and easily excreted compound, allantoin Cyberlindnera jadinii ?
-
?
uric acid + O2 + H2O purine degradation Cyberlindnera jadinii 5-hydroxyisourate + H2O2
-
?

Synonyms

Synonyms Comment Organism
Urate oxidase
-
Cyberlindnera jadinii
uricase
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Cyberlindnera jadinii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
yield of recombinant uricase is significantly improved by the combined use of a high cell-density cultivation technique and a pH control strategy of switching culture pH from 5.5 to 6.5 in the induction phase Cyberlindnera jadinii
8.5
-
assay at Cyberlindnera jadinii

pH Stability

pH Stability pH Stability Maximum Comment Organism
additional information
-
uricase activity in 50 h culture broth with pH values of 5.5 and 6.0 decreases more rapidly than that in cultures with pH values of 6.5 and 7.0, at pH 5.5, about 78% of initial uricase activity is lost within 25 h, under the same conditions, more than 85% of initial uricase activity remains in culture broth of pH 6.5 and 7.0, uricase activity in 66 h culture broth with pH 7.0 degrades much more rapidly than that in samples from 50 h culture, while for pH 6.5, the uricase is still stable, loss of uricase activity is caused by the degradation in acidic environment by proteases secreted by the host cells or releases from host cell lyses, low pH may cause instability of uricase Cyberlindnera jadinii