Similar articles

Abstract: nucleoside phosphorylase is an important enzyme involved in the biosynthesis of nucleosides. In this study, purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase were co-expressed in Escherichia coli and the intact cells were used as a catalyst for the biosynthesis of nucleosides. For protein induction, lactose was used in place of isopropyl β-D-1-thiogalactopyranoside (IPTG). When the concentration of lactose was above 0.5 mmol/L, the ability to induce protein expression was similar to that of IPTG. We determined that the reaction conditions of four bacterial strains co-expressing these genes (TUD, TAD, DUD, and DAD) were similar for the biosyntheses of 2,6-diaminopurine nucleoside and 2,6-diaminopurine deoxynucleoside. When the substrate concentration was 30 mmol/L and 0.5% of the recombinant bacterial cell volume was used as the catalyst (pH 7.5), a greater than 90% conversion yield was reached after a 2-h incubation at 50 °C. In addition, several other nucleosides and nucleoside derivatives were efficiently synthesized using bacterial strains co-expressing these recombinant enzymes.

[1]de Clercq, E., 2001. Antiviral drugs: current state of the art. J. Clin. Virol., 22(1):73-89.

[2]Esipov, R.S., Gurevich, A.I., Chuvikovsky, D.V., Chupova, L.A., Muravyova, T.I., Miroshnikov, A.I., 2002. Overexpression of Escherichia coli genes encoding nucleoside phosphorylases in the pET/Bl21(DE3) system yields active recombinant enzymes. Protein Expr. Purif., 24(1):56-60.

[3]Galmarini, C.M., Mackey, J.R., Dumontet, C., 2002. Nucleoside analogues and nucleobases in cancer treatment. Lancet Oncol., 3(7):415-423.

[4]Hamamoto, T., Okuyama, K., Noguchi, T., Midorikawa, Y., 1997a. Cloning and expression of purine nucleoside phosphorylase I gene from Bacillus stearothermophilus TH 6-2. Biosci. Biotechnol. Biochem., 61(2):272-275.

[5]Hamamoto, T., Noguchi, T., Midorikawa, Y., 1997b. Cloning of purine nucleoside phosphorylase II gene from Bacillus stearothermophilus TH 6-2 and characterization of its gene product. Biosci. Biotechnol. Biochem., 61(2):276-280.

[6]Hori, N., Uehara, K., Mikash, Y., 1992. Enzymatic synthesis of 5-methyluridine from adenosine and thymidine with high efficiency. Biosci. Biotechnol. Biochem., 56(4):580-582.

[7]Ishii, N., Shirae, H., Yokozeki, K., 1989. Enzymatic production of 5-methyluridine from purine nucleosides and thymine by Erwinia carotovora AJ-2992. Agric. Biol. Chem., 52(12):3209-3218.

[8]Kalckar, H.M., 1947. Differential spectrophotometry of purine compounds by means of specific enzymes. J. Biol. Chem., 167:429-443.

[9]Kalckar, H.M., Klenow, H., 1948. Milk xanthopterin oxidase and pteroylglutamic acid. J. Biol. Chem., 172(1):349-353.

[10]Lee, J., Filosa, S., Bonvin, J., Guyon, S., Aponte, R.A., Joanne, L., Turnbull, J.L., 2001. Expression, purification, and characterization of recombinant purine nucleoside phosphorylase from Escherichia coli. Protein Expr. Purif., 22(2):180-188.

[11]Okuyama, K., Hamamoto, T., Noguchi, T., Midorikawa, Y., 1996. Molecular cloning and expression of the pyrimidine nucleoside phosphorylase gene from Bacillus stearothermophilus TH 6-2. Biosci. Biotechnol. Biochem., 60(10):1655-1659.

[12]Okuyama, K., Shibuya, S., Hamamoto, T., Noguchi, T., 2003. Enzymatic synthesis of 2′-deoxyguanosine with nucleoside deoxyribosyltransferase-II. Biosci. Biotechnol. Biochem., 67(5):989-995.

[13]Saunders, P.P., Barbara, A.W., Saunders, G.F., 1969. Purification and comparative properties of a pyrimidine nucleoside phosphorylase from Bacillus stearothermophilus. J. Biol. Chem., 244(13):3691-3697.

[14]Takehara, M., Ling, F., Izawa, S., Inoue, Y., Kimura, A., 1995. Molecular cloning and nucleotide sequence of purine nucleoside phosphorylase and uridine phosphorylase genes from Klebsiella sp. Biosci. Biotechnol. Biochem., 59(10):1987-1990.

[15]Utagawa, T., 1999. Enzymatic preparation of nucleoside antibiotics. J. Mol. Catal. B: Enzym., 6(3):215-222.

[16]Wei, X.K., Ding, Q.B., Zhang, L., Guo, Y.L., Ou, L., Wang, C.L., 2008. Induction of nucleoside phosphorylase in Enterobacter aerogenes and enzymatic synthesis of adenine arabinoside. J. Zhejiang Univ.-Sci. B., 9(7):520-526.

[17]Yokozeki, K., Tsuji, T., 2000. A novel enzymatic method for the production of purine-2′-deoxyribonucleosides. J. Mol. Catal. B: Enzym., 10(1-3):207-213.