Literature summary for 3.2.1.23 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.

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Cloned(Commentary)

Cloned (Comment)	Organism
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

Protein Variants	Comment	Organism
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

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
YH4502	-	Pyrococcus woesei

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
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

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

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Release 2023.1 (January 2023)