4.2.2.16: levan fructotransferase (DFA-IV-forming)

This is an abbreviated version!
For detailed information about levan fructotransferase (DFA-IV-forming), go to the full flat file.

Reaction

Synonyms

2,6-beta-D-fructan D-fructosyl-D-fructosyltransferase (forming di-beta-D-fructofuranose 2,6':2',6-dianhydride), fructotransferase, levan, FTF, levan fructotransferase, levan fructotransferase (DFA-IV-forming), LFT, LFTase, LftM

ECTree

     4 Lyases

         4.2 Carbon-oxygen lyases

             4.2.2 Acting on polysaccharides

                4.2.2.16 levan fructotransferase (DFA-IV-forming)

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Results	in table
2	AA Sequence
9	Application
1	CAS Registry Number
11	Cloned(Commentary)
1	Crystallization (Commentary)
2	General Stability
23	Inhibitors
1	KM Value [mM]
8	Localization
4	Metals/Ions
6	Molecular Weight [Da]
2	Natural Substrates/ Products (Substrates)
159	Organism
16	PDB ID
9	pH Optimum
2	pH Range
9	Protein Variants
8	Purification (Commentary)
1	Reaction
1	Reaction Type
34	Reference
2	Source Tissue
4	Specific Activity [micromol/min/mg]
43	Substrates and Products (Substrate)
8	Subunits
29	Synonyms
1	Systematic Name
8	Temperature Optimum [°C]
1	Temperature Range [°C]
5	Temperature Stability [°C]

Engineering

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Engineering on EC 4.2.2.16 - levan fructotransferase (DFA-IV-forming)

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PROTEIN VARIANTS

ORGANISM

UNIPROT

COMMENTARY

LITERATURE

A264G

Microbacterium sp.

-

relative hydrolytic activity: 20.4% compared to levanbiohydrolase from Microbacterium laevaniformans. Relative hydrolytic activity of wild-type: 10.7%

693593

N238K

Microbacterium sp.

-

relative hydrolytic activity: 23.6% compared to levanbiohydrolase from Microbacterium laevaniformans. Relative hydrolytic activity of wild-type: 10.7%

693593

N85S

Microbacterium sp.

-

relative hydrolytic activity: 34.8% compared to levanbiohydrolase from Microbacterium laevaniformans. Relative hydrolytic activity of wild-type: 10.7%

693593

N85S/A264G

Microbacterium sp.

-

relative hydrolytic activity: 49.9% compared to levanbiohydrolase from Microbacterium laevaniformans. Relative hydrolytic activity of wild-type: 10.7%

693593

N85S/D268E

Microbacterium sp.

-

relative hydrolytic activity: 30% compared to levanbiohydrolase from Microbacterium laevaniformans. Relative hydrolytic activity of wild-type: 10.7%

693593

N85S/N224K/Q229E/T253N

Microbacterium sp.

-

relative hydrolytic activity: 37% compared to levanbiohydrolase from Microbacterium laevaniformans. Relative hydrolytic activity of wild-type: 10.7%

693593

P313R/F518L

Microbacterium sp.

-

relative hydrolytic activity: 15.8% compared to levanbiohydrolase from Microbacterium laevaniformans. Relative hydrolytic activity of wild-type: 10.7%

693593

D54N

Paenarthrobacter ureafaciens

Q9KJD0

inactive mutant, used for the structural analysis of the complex with sucrose

729994

additional information

Microbacterium sp.

-

residues that are critical to the intramolecular fructosyl transfer reaction of LFTase are analysed. An error-prone PCR mutagenesis process is carried out, and the mutants that led to a shift in activity from transfructosylation towards hydrolysis of levan are screened by the dinitrosalicylic acid (DNS) method. Selected mutants reveal two major products: one is a di-D-fructose-2,6':6,2'-dianhydride (DFAIV) and the other is a levanbiose. The detected levanbiose corresponds to the reaction product from LFTase lacking transferring activity N85S substitution is responsible for the strong enhancement of the hydrolytic activity

693593