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Literature summary for 1.4.1.20 extracted from

  • Tarhan, L.; Ayar-Kayali, H.
    Immobilization of phenylalanine dehydrogenase and its application in flow-injection analysis system for determination of plasma phenylalanine (2011), Appl. Biochem. Biotechnol., 163, 258-267.
    View publication on PubMed

Application

Application Comment Organism
diagnostics the immobilized enzyme is useful for establishing an analytical spectrophotometric determination method of L-Phe in the diagnostics of phenyketonuria, an inborn error of amino acid metabolism in which the conversion of L-phenylalanine to L-tyrosine is impaired and can cause profound mental retardation if not detected and treated soon after birth. Early quantitative measurement of the plasma L-Phe is essential for the diagnosis of phenylketonuria and the control of dietary therapy of the patients. The conversion efficiency of the reactor with immobillized enzyme is 100% in the range of 0.005-0.6 mM Phe at 9 mM NAD+ with a total flow rate of 0.1 mL/min. The reactor is used for the analyses of 30 samples each for 3 h per day. The half-life period of the reactor is 15 days, method evaluation, overview Sporosarcina ureae

Protein Variants

Protein Variants Comment Organism
additional information the enzyme is modified initially with 2-amino-4,6-dichloro-s-triazine, and immobilized by hexamethylenediamine and glutaraldehyde. The highest activity of immobilized PheDH is determined as 95.75 U/g support with 56% retained activity. The optimum pH value of immobilized L-PheDH was shifted from pH 10.4 to pH 11.0. pH and thermal stability of the immobilized L-PheDH are improved compared to the native enzyme Sporosarcina ureae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.038
-
L-phenylalanine pH 10.4, 30°C, immobilized enzyme Sporosarcina ureae
0.052
-
L-phenylalanine pH 10.4, 30°C, native enzyme Sporosarcina ureae
0.063
-
L-phenylalanine pH 10.4, 25°C, immobilized enzyme Sporosarcina ureae
0.076
-
L-phenylalanine pH 10.4, 30°C, immobilized enzyme Sporosarcina ureae
0.118
-
L-phenylalanine pH 10.4, 25°C, native enzyme Sporosarcina ureae
0.128
-
NAD+ pH 10.4, 25°C, immobilized enzyme Sporosarcina ureae
0.167
-
L-phenylalanine pH 10.4, 30°C, native enzyme Sporosarcina ureae
0.234
-
NAD+ pH 10.4, 25°C, native enzyme Sporosarcina ureae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-phenylalanine + H2O + NAD+ Sporosarcina ureae
-
phenylpyruvate + NH3 + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Sporosarcina ureae
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
9.1
-
native enzyme, pH 10.4, 30°C Sporosarcina ureae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-phenylalanine + H2O + NAD+
-
Sporosarcina ureae phenylpyruvate + NH3 + NADH + H+
-
r

Synonyms

Synonyms Comment Organism
L-PheDH
-
Sporosarcina ureae
PheDH
-
Sporosarcina ureae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
native enzyme Sporosarcina ureae
45 55 immobilized enzyme Sporosarcina ureae

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 45 native enzyme, 15 min, 96% activity remaining at pH 7.5, no activity remaining at pH 10.4 Sporosarcina ureae
40
-
native enzyme, 10 min, 75% activity remaining at pH 9.0 Sporosarcina ureae
50
-
native enzyme, 10 min, no activity remaining at pH 9.0 Sporosarcina ureae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.37
-
L-phenylalanine pH 10.4, 25°C, native enzyme Sporosarcina ureae
8.55
-
L-phenylalanine pH 10.4, 30°C, native enzyme Sporosarcina ureae
28.71
-
L-phenylalanine pH 10.4, 25°C, immobilized enzyme Sporosarcina ureae
49.56
-
L-phenylalanine pH 10.4, 30°C, immobilized enzyme Sporosarcina ureae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10.4
-
native enzyme Sporosarcina ureae
11
-
immobilized enzyme Sporosarcina ureae

pH Stability

pH Stability pH Stability Maximum Comment Organism
7.5
-
native enzyme, 15 min, 96% activity remaining at 30-45°C Sporosarcina ureae
9
-
native enzyme, 10 min, 75% activity remaining at 40°C, no activity at 50°C Sporosarcina ureae
10.4
-
native enzyme, 15 min, no activity remaining at 30-45°C Sporosarcina ureae

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Sporosarcina ureae
NADH
-
Sporosarcina ureae

General Information

General Information Comment Organism
malfunction an inborn error of amino acid metabolism, in which the conversion of L-phenylalanine to L-tyrosine is impaired and can cause profound mental retardation if not detected and treated soon after birth, is phenylketonuria, a disease due to an autosomal recessive inheritance that codes for a type of phenylalanine hydroxylase with reduced enzymatic activity, resulting in high levels of L-Phe in inborn fluid and phenylpyruvate in urine Sporosarcina ureae