Literature summary for 3.5.1.69 extracted from

Han, Y.B.; Wu, L.; Rich, J.R.; Huang, F.T.; Withers, S.G.; Feng, Y.; Yang, G.Y.
Comprehensive characterization of sphingolipid ceramide N-deacylase for the synthesis and fatty acid remodeling of glycosphingolipids (2015), Appl. Microbiol. Biotechnol., 99, 6715-6726.

Activating Compound

Activating Compound	Comment	Organism	Structure
Triton X-100	activates the hydrolytic reaction, best at 0.5% w/v, but inhibits the synthetic reaction	Shewanella algae

Application

Application	Comment	Organism
synthesis	the enzyme can serve as a biocatalyst in the enzymatic synthesis of glycosphingolipids since it catalyzes the reversible hydrolysis/synthesis of the amide linkage between the fatty acid and the sphingosine base in the ceramide moiety of glycosphingolipids	Pseudomonas sp.
synthesis	the enzyme can serve as a biocatalyst in the enzymatic synthesis of glycosphingolipids since it catalyzes the reversible hydrolysis/synthesis of the amide linkage between the fatty acid and the sphingosine base in the ceramide moiety of glycosphingolipids	Shewanella algae

Cloned(Commentary)

Cloned (Comment)	Organism
the gene encoding the mature SA_SCD protein, which lacks its 38-residue N-terminal secretion signal sequence and from which the 277-residue C-terminal sequence is deleted, is codon-optimized and expressed in Escherichia coli strain BL21 (DE3) pLysS as Histagged enzyme	Shewanella algae

Inhibitors

Inhibitors	Comment	Organism
Ca2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Co2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Cu2+	inhibits the hydrolytic activity and the synthetic activity of the enzyme	Shewanella algae
deoxycholic acid	complete inhibition of the hydrolytic and synthetic activity	Shewanella algae
dimethoxyethane	addition of 5% of DME strongly promoted the activity, further increasing the DME concentrations progressively inhibited synthetic activity	Shewanella algae
DMSO	low concentrations of DMSO, up to a maximum at 10%, enhance synthetic activity, while higher concentrations inhibit	Shewanella algae
Fe2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Mg2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Mn2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Ni2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
taurodeoxycholate	complete inhibition of the hydrolytic and synthetic activity	Shewanella algae
Triton X-100	activates the hydrolytic reaction, best at 0.5% w/v, but inhibits the synthetic reaction	Shewanella algae
Zn2+	inhibits the hydrolytic activity and the synthetic activity of the enzyme	Shewanella algae

Metals/Ions

Metals/Ions	Comment	Organism
Ca2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Co2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Fe2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Mg2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
Mn2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae
additional information	poor effect by EDTA	Shewanella algae
Ni2+	promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme	Shewanella algae

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas sp.	-	-	-
Shewanella algae	Q8RTY6	-	-
Shewanella algae G8	Q8RTY6	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme, lacking the secretion signal, from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography	Shewanella algae

Source Tissue

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
16-(acetylsulfanyl)hexadecanoic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
16-(acetylsulfanyl)hexadecanoic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
16-(acetylsulfanyl)hexadecanoic acid + lyso-GM1a	-	Shewanella algae G8	? + H2O	-	r
16-[(prop-2-yn-1-yl)oxy]hexadecanoic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
16-[(prop-2-yn-1-yl)oxy]hexadecanoic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
16-[(prop-2-yn-1-yl)oxy]hexadecanoic acid + lyso-GM1a	-	Shewanella algae G8	? + H2O	-	r
2-hydroxydodecanoic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
2-hydroxydodecanoic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
2-hydroxydodecanoic acid + lyso-GM1a	-	Shewanella algae G8	? + H2O	-	r
2-hydroxyhexadecanoic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
2-hydroxyhexadecanoic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
2-hydroxyhexadecanoic acid + lyso-GM1a	-	Shewanella algae G8	? + H2O	-	r
arachidic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
arachidic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
behenic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
behenic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
caproic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
caproic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
caprylic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
caprylic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
decanoic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
decanoic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
docosahexaenoic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
docosahexaenoic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
eicosapentaenoic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
eicosapentaenoic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
GM1a + H2O	-	Pseudomonas sp.	stearic acid + lyso-GM1a	-	r
GM1a + H2O	-	Shewanella algae	stearic acid + lyso-GM1a	-	r
GM3 + H2O	-	Pseudomonas sp.	stearic acid + lyso-GM3	-	r
GM3 + H2O	-	Shewanella algae	stearic acid + lyso-GM3	-	r
hexadecanedioic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
hexadecanedioic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
hexadecanedioic acid methyl ester + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
hexadecanedioic acid methyl ester + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
lauric acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
lauric acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
lignoceric acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
lignoceric acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
additional information	the enzyme catalyzes reversible reactions in which the amide linkage in glycosphingolipids is hydrolyzed or synthesized. PS_SCD also prefers glycosphingolipids possessing larger sugar moieties, but it does not show preference towards glycosphingolipids with charged head groups, the enzyme hydrolyzes sulfatide faster than GM1a and GM3, specificity of the hydrolytic and synthetic reactions of PS_SCD for hydrophilic head groups, overview	Pseudomonas sp.	?	-	?
additional information	the enzyme catalyzes reversible reactions in which the amide linkage in glycosphingolipids is hydrolyzed or synthesized. The enzyme shows high catalytic efficiency and has a very broad substrate specificity both in hydrolysis and synthesis, especially for unsaturated fatty acids and fatty acids with very short or long acyl chains. The enzyme SA_SCD hydrolyzes GM1a and GM3 fast, specificity of the hydrolytic and synthetic reactions of SA_SCD for hydrophilic head groups, overview	Shewanella algae	?	-	?
additional information	the enzyme catalyzes reversible reactions in which the amide linkage in glycosphingolipids is hydrolyzed or synthesized. The enzyme shows high catalytic efficiency and has a very broad substrate specificity both in hydrolysis and synthesis, especially for unsaturated fatty acids and fatty acids with very short or long acyl chains. The enzyme SA_SCD hydrolyzes GM1a and GM3 fast, specificity of the hydrolytic and synthetic reactions of SA_SCD for hydrophilic head groups, overview	Shewanella algae G8	?	-	?
myristic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
myristic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
oleic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
oleic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
omega-azido palmitic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
omega-azido palmitic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
omega-bromo-palmitic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
omega-bromo-palmitic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
omega-hydroxypalmitic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
omega-hydroxypalmitic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r
palmitic acid + lyso-GM1a	-	Pseudomonas sp.	? + H2O	-	r
palmitic acid + lyso-GM1a	-	Shewanella algae	? + H2O	-	r

Synonyms

Synonyms	Comment	Organism
PS_SCD	-	Pseudomonas sp.
SA_SCD	-	Shewanella algae
SCDase	-	Pseudomonas sp.
SCDase	-	Shewanella algae
sphingolipid ceramide N-deacylase	-	Pseudomonas sp.
sphingolipid ceramide N-deacylase	-	Shewanella algae

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6	-	assay at, forward direction	Pseudomonas sp.
6	-	hydrolytic activity optimum	Shewanella algae
7.5	-	assay at, reverse direction	Pseudomonas sp.
7.5	-	synthetic activity optimum	Shewanella algae

pH Range

pH Minimum	pH Maximum	Comment	Organism
5	9	activity range, hydrolysis	Shewanella algae
5.5	10	activity range, synthesis	Shewanella algae