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IUBMB Comments Phosphorylates the antibiotics hygromycin B, 1-N-hygromycin B and destomycin, but not hygromycin B2, at the 7''-hydroxy group in the destomic acid ring.
The enzyme appears in viruses and cellular organisms
Synonyms
hygromycin phosphotransferase, hygromycin b phosphotransferase,
more
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hygromycin B phosphotransferase
hygromycin phosphotransferase
phosphotransferase, hygromycin B
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hygromycin B phosphotransferase
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hygromycin B phosphotransferase
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hygromycin phosphotransferase
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hygromycin phosphotransferase
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ATP + hygromycin B = ADP + 7''-O-phosphohygromycin B
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phospho group transfer
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ATP:hygromycin-B 7''-O-phosphotransferase
Phosphorylates the antibiotics hygromycin B, 1-N-hygromycin B and destomycin, but not hygromycin B2, at the 7''-hydroxy group in the destomic acid ring.
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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resistance to hygromycin B also occurs in Escherichia coli, where a plasmid encodes a hygromycin B phosphotransferase activity that produces 4-phosphoryl-hygromycin B
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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responsible for resistance against hygromycin B, an aminocyclitol antibiotic
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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resistance to hygromycin B also occurs in Escherichia coli, where a plasmid encodes a hygromycin B phosphotransferase activity that produces 4-phosphoryl-hygromycin B
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ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
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responsible for resistance against hygromycin B, an aminocyclitol antibiotic
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Mg2+
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Mg2+
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requirement, optimal activity at 0.1 mM
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hygromycin B
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above 0.01 mM, substrate inhibition
hygromycin B
substrate inhibition
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Carcinoma, Embryonal
Generation of Mouse STO Feeder Cell Lines That Confer Resistance to Several Types of Selective Drugs.
Chagas Disease
Trypanosoma cruzi: use of herpes simplex virus-thymidine kinase as a negative selectable marker.
Diphtheria
Seamless Genome Editing in Rice via Gene Targeting and Precise Marker Elimination.
Fibrosarcoma
Measurement of unrestrained negative supercoiling and topological domain size in living human cells.
Foot-and-Mouth Disease
Agrobacterium tumefaciens-Mediated Nuclear Transformation of a Biotechnologically Important Microalga-Euglena gracilis.
Glioma
Reduced tumorigenicity of rat glioma cells in the brain when mediated by hygromycin phosphotransferase.
Herpes Simplex
Construction of strains for rapid homokaryon purification after integration of constructs at the histidine-3 ( his-3) locus of Neurospora crassa.
Herpes Simplex
Dominant positive and negative selection using a hygromycin phosphotransferase-thymidine kinase fusion gene.
Herpes Simplex
Functional identification of genes up- and down-regulated by glucocorticoids in AtT-20 pituitary cells using an enhancer trap.
Herpes Simplex
Identification of eIF2Bgamma and eIF2gamma as cofactors of hepatitis C virus internal ribosome entry site-mediated translation using a functional genomics approach.
Herpes Simplex
Nonmyeloablative immunosuppressive regimen prolongs In vivo persistence of gene-modified autologous T cells in a nonhuman primate model.
Herpes Simplex
O6-methylguanine induces intrachromosomal homologous recombination in human cells.
Herpes Simplex
Trypanosoma cruzi: use of herpes simplex virus-thymidine kinase as a negative selectable marker.
Infections
Use of Coniothyrium minitans transformed with the hygromycin B resistance gene to study survival and infection of Sclerotinia sclerotiorum sclerotia in soil.
Leukemia
Presence of a retroviral encapsidation sequence in nonretroviral RNA increases the efficiency of formation of cDNA genes.
Leukemia
Retroviral recombination rates do not increase linearly with marker distance and are limited by the size of the recombining subpopulation.
Mouth Diseases
Agrobacterium tumefaciens-Mediated Nuclear Transformation of a Biotechnologically Important Microalga-Euglena gracilis.
Newcastle Disease
[Expression and immunization testing of fusion protein of Newcastle disease virus in leaf tissue of transgenic rice]
Sarcoma
Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells.
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0.00056 - 0.03
hygromycin B
additional information
additional information
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0.0364
ATP
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0.0423
ATP
pH 7.5, 45°C, recombinant wild-type enzyme YG
0.0822
ATP
pH 7.5, 45°C, recombinant mutant enzyme HYG10
0.00056
hygromycin B
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0.0015
hygromycin B
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pH 7.5, 30°C
0.0167
hygromycin B
pH 7.5, 45°C, recombinant wild-type enzyme YG
0.03
hygromycin B
pH 7.5, 45°C, recombinant mutant enzyme HYG10
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2.5
ATP
pH 7.5, 45°C, recombinant wild-type enzyme YG
5.6
ATP
pH 7.5, 45°C, recombinant mutant enzyme HYG10
6.4
hygromycin B
pH 7.5, 45°C, recombinant wild-type enzyme YG
8
hygromycin B
pH 7.5, 45°C, recombinant mutant enzyme HYG10
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59.1
ATP
pH 7.5, 45°C, recombinant wild-type enzyme YG
68.1
ATP
pH 7.5, 45°C, recombinant mutant enzyme HYG10
267
hygromycin B
pH 7.5, 45°C, recombinant mutant enzyme HYG10
383
hygromycin B
pH 7.5, 45°C, recombinant wild-type enzyme YG
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0.61
hygromycin B
pH 7.5, 45°C, recombinant wild-type enzyme YG
1.27
hygromycin B
pH 7.5, 45°C, recombinant mutant enzyme HYG10
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7.5
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6.6 - 8.5
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pH 6.6: about 40% of maximal activity, pH 8.5: about 65% of maximal activity
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brenda
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brenda
gene hyg
UniProt
brenda
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brenda
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KHYB_STRHY
332
0
37054
Swiss-Prot
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A0A839XNL9_9PSEU
288
0
31619
TrEMBL
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A0A8I0P6I8_9ACTN
310
0
34499
TrEMBL
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A0A7X0HI31_9ACTN
307
0
34034
TrEMBL
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A0A1L7F3C8_9PSEU
328
0
35703
TrEMBL
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A0A7W8AQX8_STRST
310
0
34208
TrEMBL
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A0A840HDT2_9BRAD
319
0
35263
TrEMBL
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A0A840GN12_9BRAD
320
0
35381
TrEMBL
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A0A841BAI1_9PSEU
304
0
33786
TrEMBL
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D6U7P6_9CHLR
317
0
36439
TrEMBL
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A0A852UN05_9ACTN
320
0
35924
TrEMBL
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D3F8R8_CONWI
Conexibacter woesei (strain DSM 14684 / CIP 108061 / JCM 11494 / NBRC 100937 / ID131577)
306
0
32999
TrEMBL
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A0A7W7HUL4_9ACTN
311
0
33680
TrEMBL
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A0A1S6HYE0_9GAMM
334
0
38275
TrEMBL
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A0A839S251_9PSEU
288
0
31431
TrEMBL
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A0A7Y9MQT1_9XANT
322
0
35942
TrEMBL
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A0A7W4XYT7_KINRA
320
0
35345
TrEMBL
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A0A840S5P0_9BURK
307
0
33689
TrEMBL
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A0A7Y9RQX7_9BRAD
319
0
35263
TrEMBL
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A0A7W6U189_9BRAD
319
0
35252
TrEMBL
-
A0A7W6SRE7_9BRAD
325
0
35847
TrEMBL
-
A0A7W7WXN2_9PSEU
314
0
34858
TrEMBL
-
A0A7W9PQX7_9ACTN
310
0
34259
TrEMBL
-
A0A1D8BTP7_9PSEU
317
0
34639
TrEMBL
-
A0A7W6T0C0_9BRAD
320
0
35284
TrEMBL
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A0A8I2C3M4_BRAEL
315
0
34894
TrEMBL
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D3Q4Y8_STANL
Stackebrandtia nassauensis (strain DSM 44728 / CIP 108903 / NRRL B-16338 / NBRC 102104 / LLR-40K-21)
293
0
31768
TrEMBL
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A0A841CIL3_9PSEU
299
0
32597
TrEMBL
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41000
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1 * 41000, SDS-PAGE
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?
x * 37980, wild-type enzyme, sequence calculation, x * 38348, enzyme mutant HYG10, sequence calculation
monomer
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1 * 41000, SDS-PAGE
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L79F/S83L/G189R/E237K/S252P/D285N/L290M
random mutagenesis, mutant HYG10 contains seven substitutions and a duplication, it is thermostabilized compared to the wild-type enzyme, three-dimensional structure analysis, overview. Substitutions G189R, E237K, and D285N are located at the protein surface. The mutant shows altered kinetics at 45°C compared to the wild-type enzyme
additional information
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use the hygromycin phosphotransferase gene (hpt) as a selective marker gene for tracking plastid transformation in rice. A site-specific transformation vector of rice plastid, pCTE04, is constructed for dicistronic expression of the hygromycin phosphotransferase (hpt) and enhanced green fluorescent protein genes (EGFP). Transplastomic rice plants successfully grow up after a gradual selection of hygromycin resistance. Integration of the dicistronic expression cassette into the intergenic region trnI-trnA of plastid genome is verified by Southern blotting analysis. Transplastomic expression of the hpt and EGFP genes is identified by RT-PCR. The stable expression of dicistronic cassette in the rice is confirmed by confocal microscopy
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35
T1/2 of wild-type enzyme HYG
48 - 74
T1/2 of mutant enzyme HYG10, Tmax is 74°C
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glycerol and bovine serum albumin stabilize
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-20°C, in 50% v/v glycerol and 0.1% bovine serum albumin, stable for up to 6 months
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recombinant His6-tagged enzyme fom Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration
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gene hyg, subcloning in Escherichia coli strain JM109, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), recombinant enzyme expression from plasmid pT8S-P31 in Thermus thermophilus strain B27 TH104 providing hygromycin B resistance up to 60°C, while recombinant expression of a mutant variant HYG10, containing seven substitutions and a duplication, provides HgB resistance up to 74°C
recombinant gene expression in transgenic Oryza sativa from site-specific transformation vector of rice plastid, pCTE04
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biotechnology
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use the hygromycin phosphotransferase gene (hpt) as a selective marker gene for tracking plastid transformation in rice (Oryza sativa)
molecular biology
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use the hygromycin phosphotransferase gene (hpt) as a selective marker gene for tracking plastid transformation in rice (Oryza sativa)
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Zalacain, M.; Pardo, J.M.; Jimenez, A.
Purification and characterization of a hygromycin B phosphotransferase from Streptomyces hygroscopicus
Eur. J. Biochem.
162
419-422
1987
Streptomyces hygroscopicus
brenda
Pardo, J.M.; Malpartida, F.; Rico, M.; Jimenez, A.
Biochemical basis of resistance to hygromycin B in Streptomyces hygroscopicus - the producing organism
J. Gen. Microbiol.
131
1289-1298
1985
Streptomyces hygroscopicus
brenda
Sugimoto, N.; Takakura, Y.; Shiraki, K.; Honda, S.; Takaya, N.; Hoshino, T.; Nakamura, A.
Directed evolution for thermostabilization of a hygromycin B phosphotransferase from Streptomyces hygroscopicus
Biosci. Biotechnol. Biochem.
77
2234-2241
2013
Streptomyces hygroscopicus (P09979), Streptomyces hygroscopicus
brenda
Li, D.; Tang, N.; Liu, M.; Shen, C.; Hu, Y.; Xia, Y.; Cao, M.
Using hygromycin phosphotransferase and enhanced green fluorescent protein genes for tracking plastid transformation in rice (Oryza sativa L.) via gold particle bombardment
Nanosci. Nanotechnol. Lett.
8
409-417
2016
Streptomyces sp.
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brenda
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