Table 4.

Conditions for performing the polyenzymatic cascades of nucleoside synthesis with recombinant thermophilic and mesophilic enzymes with various D-pentoses.

Nucleoside	2ClAde Conc., mM	ATP Conc., mM	D-Pentose Conc.	Enzyme Ratio, Units			Max. Nucleoside Content, %
				RK	PPM	PNP
Thermus thermophilus
Ribo-2ClAde	0.34	2.5	2 mM D-ribose	0.3 *	0.1	0.5 **	78 (30 min)
dRibo-2ClAde			2 mM 2-deoxy-D-ribose	0.3 *	0.1	0.5 **	61 (30 min)
Ara-2ClAde			60 mM D-arabinose	11 *	1	10 **	52 (4 h)
Xylo-2ClAde			60 mM D-xylose	11 *	1	10 **	4 (24 h)
Clofarabine			10 mM 2-deoxy-2-fluoro-D-arabinose	6 *	1	10 **	19 (24 h)
Escherichia coli
Ribo-2ClAde	0.34	2.5	2 mM D-ribose	0.2	0.1	0.4	91 (1 h)
dRibo-2ClAde			2 mM 2-deoxy-D-ribose	0.2	0.1	0.4	73 (30 min)
Ara-2ClAde			60 mM D-arabinose	5	1	9	67 (1 h)
Xylo-2ClAde			60 mM D-xylose	5	1	9	26 (24 h)
Clofarabine			10 mM 2-deoxy-2-fluoro-D-arabinose	6	1	4	46 (24 h)

* Ribokinase from Thermus species 2.9 (TspRK, gene QT17_05185) [23] was used. ** Purine nucleoside phosphorylase II (TthPNPII) was used.