Literature summary for 6.5.1.3 extracted from

Blondal, T.; Thorisdottir, A.; Unnsteinsdottir, U.; Hjorleifsdottir, S.; Aevarsson, A.; Ernstsson, S.; Fridjonsson, O.H.; Skirnisdottir, S.; Wheat, J.O.; Hermannsdottir, A.G.; Sigurdsson, S.T.; Hreggvidsson, G.O.; Smith, A.V.; Kristjansson, J.K.
Isolation and characterization of a thermostable RNA ligase 1 from a Thermus scotoductus bacteriophage TS2126 with good single-stranded DNA ligation properties (2005), Nucleic Acids Res., 33, 135-142.

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Application

Application	Comment	Organism
molecular biology	RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE)	Phage TS2126
molecular biology	the properties of TS2126 RNA ligase 1 makes it very attractive for processes like adaptor ligation, and single-stranded solid phase gene synthesis	Phage TS2126

Cloned(Commentary)

Cloned (Comment)	Organism
RNA ligase gene is inserted into a pET23b expression vector with a C-terminal His6-tag. This vector is transformed into Escherichia coli strain BL21	Phage TS2126

Inhibitors

Inhibitors	Comment	Organism	Structure
ATP	above 2.5 mM, complete inhibition at 10 mM. The inhibition can be partially rescued by adding more Mg2+ to the reaction	Phage TS2126

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.007	-	ATP	activity with RNA	Phage TS2126
0.04	-	ATP	activity with DNA	Phage TS2126

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	activates, optimum: 5-10 mM	Phage TS2126
Mn2+	activates, showing slightly higher activity than Mg2+	Phage TS2126

Organism

Organism	UniProt	Comment	Textmining
Phage TS2126	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Phage TS2126

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + (ribonucleotide)n + (ribonucleotide)m	the TS2126 RNA ligase catalyzes both inter- and intra-molecular single-stranded DNA ligation to more than 50% completion in not more than a few hours at an elevated temperature, although favoring intramolecular ligation on RNA and single-stranded DNA substrates	Phage TS2126	AMP + diphosphate + (ribonucleotide)n+m	-	?

Synonyms

Synonyms	Comment	Organism
RNA ligase 1	-	Phage TS2126

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
65	70	-	Phage TS2126

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
50	75	50°C: about 35% of maximal activity, 75°C: about 60% of maximal activity	Phage TS2126

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
65	-	realatively stable at	Phage TS2126
80	-	loses all activity above	Phage TS2126

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	8	MOPS buffer	Phage TS2126

pH Range

pH Minimum	pH Maximum	Comment	Organism
7	8.5	pH 7.0: about 40% of maximal activity, pH 8.5: about 30% of maximal activity	Phage TS2126

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Release 2023.1 (January 2023)