Literature summary for 3.5.1.69 extracted from

Kuchar, L.; Rotkova, J.; Asfaw, B.; Lenfeld, J.; Horak, D.; Korecka, L.; Bilkova, Z.; Ledvinova, J.
Semisynthesis of C17:0 isoforms of sulphatide and glucosylceramide using immobilised sphingolipid ceramide N-deacylase for application in analytical mass spectrometry (2010), Rapid Commun. Mass Spectrom., 24, 2393-2399.

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Application

Application	Comment	Organism
analysis	the enzyme is immobilised on magnetic macroporous cellulose and is used to semisynthesise C17:0 glucosylceramide and C17:0 sulfatide, which are required as internal standards for quantification of the corresponding glycosphingolipids by tandem mass spectrometry	Pseudomonas sp. TK4
synthesis	the enzyme is immobilised on magnetic macroporous cellulose and is used to semisynthesise C17:0 glucosylceramide and C17:0 sulfatide, which are required as internal standards for quantification of the corresponding glycosphingolipids by tandem mass spectrometry. A high rate of conversion is achieved for both lipids, 80% for C17:0 sulfatide and 90% for C17:0 glucosylceramide. In contrast to synthesis with a soluble form of the enzyme, use of immobilised SCDase significantly reduces the contamination of the sphingolipid products with other isoforms, so further purification is not necessary	Pseudomonas sp. TK4

General Stability

General Stability	Organism
SCDase immobilised on magnetic macroporous cellulose maintains its activity without loss after 15 uses	Pseudomonas sp. TK4

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas sp. TK4	-	commercial preparation	-

Reaction

Reaction	Comment	Organism	Reaction ID
ganglioside GM2 + H2O = lyso-GM2-ganglioside + octadecanoic acid	in addition to its primary deacylation function, this enzyme is able to reacylate lyso-sphingolipids under specific conditions	Pseudomonas sp. TK4	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
commercial preparation	-	Pseudomonas sp. TK4	-

Storage Stability

Storage Stability	Organism
4°C, 50 mM phosphate buffer, pH 7, 0.1% Triton X-100, long-term stability is achieved by immobilisation on magnetic macroporous cellulose. The enzyme retains the same activity after 1.5 years in storage buffer	Pseudomonas sp. TK4

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
gangliotetraosylceramide + H2O	immobilised enzyme, pH 5, 37°C. The rate of hydrolysis of the substrate by the immobilised enzyme is 62% after 20 h as compared to 48% with the soluble enzyme under the same reaction conditions	Pseudomonas sp. TK4	glucosylceramide	using the soluble form of the SCDase C17:0 glucosylceramide is prepared in bulk solution. Mass spectra reveal a considerable amount of contaminants in the final product. The contaminants are identified as glucosylceramide isoforms with C16:0 and C18:0 fatty acids. The total yield of glucosylceramide is 99%. Products are analyzed by HPTLC	?

Synonyms

Synonyms	Comment	Organism
SCDase	-	Pseudomonas sp. TK4
sphingolipid ceramide N-acylase	-	Pseudomonas sp. TK4

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Pseudomonas sp. TK4

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5	-	assay at	Pseudomonas sp. TK4

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Release 2023.1 (January 2023)