Abstract
ATP sulfurylase (ATPS, C 2.7.7.4) reversibly catalyzes the reaction between ATP and sulfate to produce APS and pyrophosphate (PPi), and has been used in pyrosequencing. The gene coding ATP sulfurylase was amplified from the genomic DNA of Saccharomyces cerevisias (CICC 1202), and cloned into prokaryotic expression plasmid pET28a(+) to provide a recombinant expression plasmid pET28a(+)-ATPS. Upon IPTG induction, ATP sulfurylase was produced by E. coli BL21(DE3) harboring the recombinant expression plasmid pET28a(+)-ATPS. The relative molecular weight of recombinant ATP sulfurylase with His tag was about 60 kD. Including the extra desalting step, only two purification steps are required to obtain the recombinant ATPS with electrophoretic pure grade. The specific activity of the purified recombinant ATP sulfurylase was as high as 5.1 × 104 U/mg. The successful application of the enzyme in pyrosequencing was also demonstrated.
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References
Renosto F, Martin RL, Borrell JL et al (1991) ATP sulfurylase from trophosome tissue of Riftia pachyptila (hydrothermal vent tube worm). Arch Biochem Biophys 290:66–78
Hanna E, MacRae IJ, Medina DC et al (2002) ATP sulfurylase from the hyperthermophilic chemolithotroph Aquifex aeolicus. Arch Biochem Biophys 406:275–288
Renosto F, Martin RL, Wailes LM et al (1990) Regulation of inorganic sulfate activation in filamentous fungi. Allosteric inhibition of ATP sulfurylase by 3'-phosphoadenosine-5'-phosphosulfate. J Biol Chem 265:10300–10308
Yu M, Martin RL, Jain S et al (1989) Rat liver ATP-sulfurylase: purification, kinetic characterization, and interaction with arsenate, selenate, phosphate, and other inorganic oxyanions. Arch Biochem Biophys 269:165–174
Osslund T, Chandler C, Segel IH (1982) ATP sulfurylase from higher plants: purification and preliminary kinetics studies on the cabbage leaf enzyme. Plant Physiol 70:39–45
Leyh TS, Suo Y (1992) GTPase-mediated activation of ATP sulfurylase. J Biol Chem 267:542–545
Foster BA, Thomas SM, Mahr JA et al (1994) Cloning and sequencing of ATP sulfurylase from Penicillium chrysogenum. Identification of a likely allosteric domain. J Biol Chem 269:19777–19786
Li H, Deyrup A, Mensch JR et al (1995) The isolation and characterization of cDNA encoding the mouse bifunctional ATP sulfurylase-adenosine 5′-phosphosulfate kinase. J Biol Chem 270:29453–29459
Klonus D, Riesmeier JW, Willmitzer L (1995) A cDNA clone for an ATP-sulfurylase from Arabidopsis thaliana. Plant Physiol 107:653–654
Nyren P, Nore BF, Baltscheffsky M (1986) Studies on photosynthetic inorganic pyrophosphate formation in Rhodospirillum rubrum chromatophores. Biochim Biophys Acta 851:276–282
Nyren P, Karamouhamed S, Ronagshi M (1996) Real-time sequence-based DNA analyses using bioluminescence. In: Hastings et al (ed) Bioluminescence and chemiluminescence molecular reporting with photons, p 466–469
Ahmadian A, Ehn M, Hober S (2006) Pyrosequencing: history, biochemistry and future. Clin Chim Acta 363:83–94
Nyren P (1987) Enzymatic method for continuous monitoring of DNA polymerase activity. Anal Biochem 167:235–238
Cherest H, Kerjan P, Surdin-Kerjan Y (1987) The Saccharomyces cerevisiae MET 3 gene: nucleotide sequence and relationship of the 5′ non-coding region to that of MET25. Mol Gen Genet 210:307–313
Karamohamed S, Nyren P (1999) Real-time detection and quantification of adenosine triphosphate sulfurylase activity by a bioluminometric approach. Anal Biochem 271:81–85
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Luo, J., Wu, W., Zou, B., Song, Q., Zhou, G. (2016). Expression and Purification of ATP Sulfurylase from Saccharomyces cerevisias in Escherichia coli and Its Application in Pyrosequencing. In: Zhou, G., Song, Q. (eds) Advances and Clinical Practice in Pyrosequencing. Springer Protocols Handbooks. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3308-2_16
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DOI: https://doi.org/10.1007/978-1-4939-3308-2_16
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