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Literature summary extracted from

  • Xiong, A.S.; Peng, R.H.; Zhuang, J.; Li, X.; Xue, Y.; Liu, J.G.; Gao, F.; Cai, B.; Chen, J.M.; Yao, Q.H.
    Directed evolution of a beta-galactosidase from Pyrococcus woesei resulting in increased thermostable beta-glucuronidase activity (2007), Appl. Microbiol. Biotechnol., 77, 569-578.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.23 gene hlacz-sh, chemical synthesis of the gene derived from Pyrococcus woesei for direct evolution, DNA and amino acid sequence determination and anaylsis, library construction, expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta-Blue (DE3) Pyrococcus woesei

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.23 T29A/V213I/L217M/N277H/I387V/R491C/N496D large range mutant screening after direct chemical evolution, the mutant variant YG6762 has nucleotide changes in 13 sites: T30C, A85G, C621A, G637A, T649A, A829V, T912G, C924T, A1159G, G1242C, C1471T, A1486G, and C1497A, the mutant YG6762 shows 4.9-7.5fold increased activity, dependent on the reaction temperature, but decreased thermostability, compared to the wild-type enzyme, overview, revertion of the mutation at each single of the 8 mutation sites results in decreased activity of the altered mutant, especially when reverting residue 277 to Asn, overview Pyrococcus woesei

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.23 Pyrococcus woesei O52629 wild-type gene hlacz-sh; gene hlacz-sh
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.23 2-nitrophenyl beta-D-galactopyranoside + H2O
-
 Pyrococcus woesei 2-nitrophenol + beta-D-galactose
-
 ?
3.2.1.23 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Pyrococcus woesei 4-nitrophenol + beta-D-galactose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.23 More three-dimensional structure analysis and molecular modeling, overview Pyrococcus woesei

Synonyms

EC Number Synonyms Comment Organism
3.2.1.23 YH4502
-
 Pyrococcus woesei

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.23 90
-
-
 Pyrococcus woesei

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.23 100
-
 the mutant enzyme YG6762 shows 40% remaining activity after 1 h, and 30% remaining activity after 2 h, wild-type enzyme YH4502 shows over 75% remaining actiivty after 1 h, and more than 50% remiaing activity after 2 h Pyrococcus woesei

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.23 4.5
-
 mutant YG6762 Pyrococcus woesei
3.2.1.23 5.5
-
 wild-type enzyme YH4502 Pyrococcus woesei