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Information on EC 2.7.7.64 - UTP-monosaccharide-1-phosphate uridylyltransferase and Organism(s) Arabidopsis thaliana and UniProt Accession Q9C5I1
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2.7.7.64 UTP-monosaccharide-1-phosphate uridylyltransferaseSpecify your search results
Word Map
- 2.7.7.64
- udp-sugars
-
gal-1-p
- synthesis
- udp-glucose
- udp-glca
- 1-phosphates
-
co-substrate
-
phytohormones
- l-arabinose
- glycolipids
-
three-enzyme
-
pisum
- sativum
- molecular biology
The taxonomic range for the selected organisms is: Arabidopsis thaliana
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Synonyms
atusp, psusp, uspase, udp-galactose/glucose pyrophosphorylase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
AtUSP
PATHWAY SOURCE
PATHWAYS
KEGG
-
-, -, -, -, -, -
CAS REGISTRY NUMBER
COMMENTARY
223918-15-8
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-galactose 1-phosphate + UTP
UDP-galactose + diphosphate
-
-
-
?
D-glucose 1-phosphate + UTP
UDP-glucose + diphosphate
pH 7, 35°C, 2 mM Mg2+
reaction stop by boiling, analysis of products by HPLC (monitoring at 262 nm)
-
r
D-glucuronic acid 1-phosphate + UTP
UDP-glucuronic acid + diphosphate
-
-
-
?
L-arabinose 1-phosphate + UTP
UDP-L-arabinose + diphosphate
-
-
-
?
N-acetylglucosamine 1-phosphate + UTP
UDP-N-acetylglucosamine + diphosphate
poorly catalysed reaction
-
-
?
UDP-galacturonic acid + diphosphate
galacturonic acid 1-phosphate + UTP
reduced activity in floret extracts depleted for AtUSP by immunprecipitation: only endogenous UDP-galacturonic acid-utilizing pyrophosphorylase activity and final step in myo-inositol oxidation pathway
-
-
?
UDP-monosaccharide + diphosphate
monosaccharide 1-phosphate + UTP
salvage pathway
-
-
r
UTP + alpha-D-glucose 1-phosphate
diphosphate + UDP-alpha-D-glucose
-
-
-
r
UTP + alpha-D-glucuronate 1-phosphate
diphosphate + UDP-alpha-D-glucuronate
-
-
-
r
4-azido-D-galactose 1-phosphate + UTP
UDP-4-azido-D-galactose + diphosphate
-
-
43% conversion rate
-
?
6-deoxy-D-galactose 1-phosphate + UTP
UDP-6-deoxy-D-galactose + diphosphate
-
-
29% conversion rate
-
?
D-galactose 1-phosphate + UTP
UDP-D-galactose + diphosphate
-
-
95% conversion rate
-
?
D-glucose 1-phosphate + UTP
UDP-D-glucose + diphosphate
-
-
28% conversion rate
-
?
UTP + monosaccharide 1-phosphate
diphosphate + UDP-monosaccharide
-
the enzyme has a critical role in pollen development. The products of the AtUSP reaction can act as precursors for the synthesis of glycolipids, glycoproteins, and cell wall components including pectin and hemicellulose
-
-
?
additional information
?
-
no activity detectable for substrates L-fucose 1-phosphate and mannose 1-phosphate
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
UDP-galacturonic acid + diphosphate
galacturonic acid 1-phosphate + UTP
reduced activity in floret extracts depleted for AtUSP by immunprecipitation: only endogenous UDP-galacturonic acid-utilizing pyrophosphorylase activity and final step in myo-inositol oxidation pathway
-
-
?
UDP-monosaccharide + diphosphate
monosaccharide 1-phosphate + UTP
salvage pathway
-
-
r
UTP + alpha-D-glucose 1-phosphate
diphosphate + UDP-alpha-D-glucose
-
-
-
r
UTP + alpha-D-glucuronate 1-phosphate
diphosphate + UDP-alpha-D-glucuronate
-
-
-
r
UTP + monosaccharide 1-phosphate
diphosphate + UDP-monosaccharide
-
the enzyme has a critical role in pollen development. The products of the AtUSP reaction can act as precursors for the synthesis of glycolipids, glycoproteins, and cell wall components including pectin and hemicellulose
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Mn2+
additional information
the enzyme requires bivalent cations for activity. HvUSP accepts other bivalent cation such as Ca2+, Mn2+, and Zn2+, but prefers Mg2+
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
8.14
dTTP
-
pH not specified in the publication, temperature not specified in the publication
3.74
dUTP
-
pH not specified in the publication, temperature not specified in the publication
2.88
UTP
-
pH not specified in the publication, temperature not specified in the publication
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
27
dTTP
-
pH not specified in the publication, temperature not specified in the publication
219
dUTP
-
pH not specified in the publication, temperature not specified in the publication
348
UTP
-
pH not specified in the publication, temperature not specified in the publication
additional information
D-galactose 1-phosphate
Kcat/KM: 490000 1/M/sec, pH 7, 35°C
additional information
D-glucose 1-phosphate
Kcat/KM: 530000 1/M/sec, pH 7, 35°C
additional information
D-glucuronic acid 1-phosphate
Kcat/KM: 710000 1/M/sec, pH 7, 35°C
additional information
D-xylose 1-phosphate
Kcat/KM: 51000 1/M/sec, pH 7, 35°C
additional information
diphosphate
Kcat/KM: 860000 1/M/sec, pH 7, 35°C
additional information
L-arabinose 1-phosphate
Kcat/KM: 92000 1/M/sec, pH 7, 35°C
additional information
UDP-glucose
Kcat/KM: 480000 1/M/sec, pH 7, 35°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3.3
dTTP
-
pH not specified in the publication, temperature not specified in the publication
59
dUTP
-
pH not specified in the publication, temperature not specified in the publication
121
UTP
-
pH not specified in the publication, temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
143-255% UDP-L-arabinose synthesising activity in extracts from transgenic plants (aerial parts) compared to wild-type plant
additional information
25% of wild-type UDP-L-arabinose synthesising activity in plant extracts (aerial parts) upon repression of AtUSP gene expression
additional information
no activity detectable for substrates L-fucose 1-phosphate and mannose 1-phosphate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
45
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
AtUSP mRNA-specific qRT-PCR, strong expression in vascular tissue (promoter: beta-glucuronidase assay)
relatively high levels in most cell types (AtUSP mRNA-specific qRT-PCR)
young leaves (AtUSP mRNA-specific qRT-PCR), strong expression in vascular tissue (promoter: beta-glucuronidase assay)
AtUSP mRNA-specific qRT-PCR, strong expression in vascular tissue (promoter: beta-glucuronidase assay), essential for pollen development since loss-of-function mutation in enzyme causes male infertility
strong expression in growing region (promoter: beta-glucuronidase assay), young roots (AtUSP mRNA-specific qRT-PCR)
young seedlings, expression in most cell types (AtUSP mRNA-specific qRT-PCR)
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
USP1 and USP2, differential centrifugation and Western blot
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
gene silencing of USP by miRNA causes a concomitant reduction of USP and of glucuronokinase activity presumably to prevent the accumulation of sugar-1-phosphates interfering with normal metabolism and depleting the phosphate pool of the cell
malfunction
-
A knock-out of the USP gene results in non-fertile pollen. Mutant plants show an arabinose reduction in the cell wall, and accumulate mainly two sugars, arabinose and xylose, in the cytoplasm
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). USP_ARATH
614
0
67852
Swiss-Prot
Secretory Pathway (Reliability: 5)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
70000
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
functional UDP-sugar diphosphorylase from Arabidopsis thaliana, constitutively expressed in Pichia pastoris and secreted into the extracellular medium, is used for synthesis of UDP-alpha-D-glucuronate, purification of the UDP-sugar from medium by anion exchange chromatography. Purification and assay methods optimization, overview. The fermentation of Pichia pastoris strain SMD1168H-C-AtUSP is performed at 30°C and pH 7.0 throughout the procedure, 20% glucose is supplemented at a rate of 9 ml/h, and DO is maintained at over 20%. After 60 h, the cell wet weight stabilizes (a wet weight of 276.7 g/l is obtained at 96 h). The crude enzyme activity increases up to 84 h, then it is stabilized, with activity 1601 U/ml. Thus, USP activity increases in 5-l fermenter compared with that in shaken flasks. The optimal temperature is around 35°C, lower than the optimal temperature of enzymatic reaction by purified AtUSP (45°C), while the optimal pH value and metal ion conditions are close to those of the purified enzyme system
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
chelating chromatography (elution: 250 mM imidazole) followed by cleavage of N-terminal thioredoxin- and His-tag with thrombin and DEAE Sepharose FF chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
from cDNA generated from total RNA from 2-week-old seedlings in pGEM T-Easy for amplification and in pET32a for inducible expression with N-terminal thioredoxin- and His-tag in Escherichia coli BL21(DE3) and in pBI121 for generation of transgenic plants via Agrobacterium tumefaciens
gene USP, functional UDP-sugar diphosphorylase from Arabidopsis thaliana is constitutively expressed in Pichia pastoris strain SMD1168H and secreted into the extracellular medium using the secretion signal from Saccharomyces cerevisiae alpha-factor prepro-peptide
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
molecular biology
polyclonal rabbit anti-AtUSP antibody recognizes USP1 and USP2
synthesis
in mammals, uridine 5'-diphosphate-glucose (UDP-Glc) and uridine 5'-diphosphate-glucuronic acid (UDP-GlcA) are common building blocks of glycans and glycoconjugates. The commercial demand for these high-energy donors is increasing. To produce valuable UDP-GlcA in a cost-effective way, UDP-sugar pyrophosphorylase from Arabidopsis thaliana is constitutively expressed in Pichia pastoris and secreted into the extracellular medium. The synthesis of 4.2 g UDP-GlcA or 5.5 g UDP-Glc per liter of culture is revealed in the culture medium, without any need for purification. An anion exchange chromatography purification method for UDP-sugars is also developed. This route opens a door to large-scale production of the cheaper UDP-GlcA
synthesis
-
use of UDP-sugar pyrophosphorylase (USP) from Arabidopsis thaliana with a galactokinase from Streptococcus pneumoniae TIGR4 (SpGalK) and an inorganic diphosphatase (PPase) to effectively synthesize UDP-sugars
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Schnurr, J.A.; Storey, K.K.; Jung, H.J.; Somers, D.A.; Gronwald, J.W.
UDP-sugar pyrophosphorylase is essential for pollen development in Arabidopsis
Planta
224
520-532
2006
Arabidopsis thaliana
Kotake, T.; Hojo, S.; Yamaguchi, D.; Aohara, T.; Konishi, T.; Tsumuraya, Y.
Properties and physiological functions of UDP-sugar pyrophosphorylase in Arabidopsis
Biosci. Biotechnol. Biochem.
71
761-771
2007
Arabidopsis thaliana (Q9C5I1)
Gronwald, J.W.; Miller, S.S.; Vance, C.P.
Arabidopsis UDP-sugar pyrophosphorylase: Evidence for two isoforms
Plant Physiol. Biochem.
46
1101-1105
2008
Arabidopsis thaliana (Q9C5I1)
Siddique, S.; Endres, S.; Atkins, J.M.; Szakasits, D.; Wieczorek, K.; Hofmann, J.; Blaukopf, C.; Urwin, P.E.; Tenhaken, R.; Grundler, F.M.; Kreil, D.P.; Bohlmann, H.
Myo-inositol oxygenase genes are involved in the development of syncytia induced by Heterodera schachtii in Arabidopsis roots
New Phytol.
184
457-472
2009
Arabidopsis thaliana (Q9C5I1)
Geserick, C.; Tenhaken, R.
UDP-sugar pyrophosphorylase is essential for arabinose and xylose recycling, and is required during vegetative and reproductive growth in Arabidopsis
Plant J.
74
239-247
2013
Arabidopsis thaliana
Liu, J.; Zou, Y.; Guan, W.; Zhai, Y.; Xue, M.; Jin, L.; Zhao, X.; Dong, J.; Wang, W.; Shen, J.; Wang, P.G.; Chen, M.
Biosynthesis of nucleotide sugars by a promiscuous UDP-sugar pyrophosphorylase from Arabidopsis thaliana (AtUSP)
Bioorg. Med. Chem. Lett.
23
3764-3768
2013
Arabidopsis thaliana
Geserick, C.; Tenhaken, R.
UDP-sugar pyrophosphorylase controls the activity of proceeding sugar-1-kinases enzymes
Plant Signal. Behav.
8
e25478
2013
Arabidopsis thaliana (Q9C5I1)
Zang, L.; Du, R.; Zang, H.; Wang, F.; Sheng, J.
Production of Arabidopsis thaliana UDP-sugar pyrophosphorylase by Pichia pastoris and its application in efficient UDP-Glucose and UDP-glucuronic acid synthesis
Appl. Biochem. Microbiol.
55
631-638
2019
Arabidopsis thaliana (Q9C5I1)
-
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