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Literature summary for 1.23.1.1 extracted from

  • Xiao, Y.; Shao, K.; Zhou, J.; Wang, L.; Ma, X.; Wu, D.; Yang, Y.; Chen, J.; Feng, J.; Qiu, S.; Lv, Z.; Zhang, L.; Zhang, P.; Chen, W.
    Structure-based engineering of substrate specificity for pinoresinol-lariciresinol reductases (2021), Nat. Commun., 12, 2828.
    View publication on PubMed
Search for more literature for 1.23.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta (DE3) cells Arabidopsis thaliana
expressed in Escherichia coli Rosetta (DE3) cells Isatis tinctoria

Crystallization (Commentary)

Crystallization (Comment) Organism
enzyme in apo, substrate-bound and product-bound states, sitting drop vapor diffusion method, using 0.2 M sodium citrate tribasic, 0.1 M sodium citrate/citric acid, pH 4.0 and 20% (w/v) polyethylene glycol (PEG) 3350 Isatis tinctoria
enzyme PrR2 in apo, substrate-bound and product-bound states, sitting drop vapor diffusion method, using 0.2 M magnesium chloride, 0.1 M sodium HEPES, pH 7.5 and 25% (w/v) PEG 3350 for the apoenzyme, and using 2.1 M DL-malic acid, pH 7.0 for the substrate-bound states Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
N98S the mutant of isoform PrR2 shows somewhat increased conversion rate of lariciresinol and significantly reduced activity with secoisolariciresinol as compared to the wild type enzyme Arabidopsis thaliana
S98A the mutant somewhat enhances the conversion rate of lariciresinol (more than 40% conversion rate) while significantly reduces that of secoisolariciresinol as compared to the wild type enzyme Isatis tinctoria
S98H the mutant somewhat enhances the conversion rate of lariciresinol (more than 40% conversion rate) while significantly reduces that of secoisolariciresinol as compared to the wild type enzyme Isatis tinctoria
S98N the mutant somewhat enhances the conversion rate of lariciresinol while significantly reduces that of secoisolariciresinol as compared to the wild type enzyme Isatis tinctoria
V46A the mutant somewhat enhances the conversion rate of lariciresinol (more than 40% conversion rate) while significantly reduces that of secoisolariciresinol as compared to the wild type enzyme Isatis tinctoria
V46A the mutant of isoform PrR2 shows significantly reduced conversion rate of lariciresinol and no activity with secoisolariciresinol as compared to the wild type enzyme Arabidopsis thaliana
V46L the mutant somewhat enhances the conversion rate of lariciresinol while significantly reduces that of secoisolariciresinol as compared to the wild type enzyme Isatis tinctoria
V46L the mutant of isoform PrR2 shows somewhat increased conversion rate of lariciresinol and no activity with secoisolariciresinol as compared to the wild type enzyme Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0294
-
 (+/-)-pinoresinol mutant enzyme V46A, at pH 7.0 and 37°C Isatis tinctoria

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(+)-pinoresinol + NADPH + H+ Isatis tinctoria
-
 (+)-lariciresinol + NADP+
-
 ?
(-)-pinoresinol + NADPH + H+ Arabidopsis thaliana
-
 (-)-lariciresinol + NADP+
-
 ?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q9SVP6
-
-
Isatis tinctoria I6LRS1
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Superdex 200 gel filtration Arabidopsis thaliana
Ni-NTA column chromatography and Superdex 200 gel filtration Isatis tinctoria

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(+)-pinoresinol + NADPH + H+
-
 Isatis tinctoria (+)-lariciresinol + NADP+
-
 ?
(+/-)-pinoresinol + NADPH + H+
-
 Isatis tinctoria (+/-)-lariciresinol + NADP+
-
 ?
(+/-)-pinoresinol + NADPH + H+ the enzyme has a substrate preference for pinoresinol Arabidopsis thaliana (+/-)-lariciresinol + NADP+
-
 ?
(-)-pinoresinol + NADPH + H+
-
 Arabidopsis thaliana (-)-lariciresinol + NADP+
-
 ?
additional information isoform PrR2 does not use (+)-lariciresinol as substrate Arabidopsis thaliana ?
-
 ?

Synonyms

Synonyms Comment Organism
pinoresinol-lariciresinol reductase
-
 Isatis tinctoria
PLR1
-
 Isatis tinctoria
PrR2 isoform Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.91
-
 (+/-)-pinoresinol mutant enzyme V46A, at pH 7.0 and 37°C Isatis tinctoria

Cofactor

Cofactor Comment Organism Structure
NADPH
-
 Isatis tinctoria
NADPH
-
 Arabidopsis thaliana

General Information

General Information Comment Organism
physiological function the enzyme is involved in the lignan biosynthesis Isatis tinctoria
physiological function the enzyme is involved in the lignan biosynthesis Arabidopsis thaliana

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
64.7
-
 (+/-)-pinoresinol mutant enzyme V46A, at pH 7.0 and 37°C Isatis tinctoria