2.4.2.6

K+

K+-activated type II enzyme: binding of K+ is required to enhance activity through induced conformational change

Lactococcus lactis

2.4.2.6

does not require phosphate derivatives such as ATP, thiamine diphosphate, pyridoxal phosphate to complete 2-deoxyribose transfer

Lactococcus lactis

2.4.2.6

in pET-22b(+) for expression in Escherichia coli BL21(DE3); in pGEM-T for cloning

Lactococcus lactis

2.4.2.6

Ca2+

inhibits base exchange activity at 1 mM

Lactococcus lactis

2.4.2.6

CO2

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

Cu2+

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

Mg2+

inhibits base exchange activity at 1mM

Lactococcus lactis

2.4.2.6

Mn2+

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

Zn2+

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

K+

enhances base exchange activity and thermostability, 1.66-fold activation by 1 mM KCl (reaches plateau at 20 mM)

Lactococcus lactis

2.4.2.6

enzymatic activity depends on monovalent cations as cofactors rather than catalysts; K+ and NH4+ have no synergistic effects; Li+ and Na+ do not activate

Lactococcus lactis

2.4.2.6

NH4+

enhances base exchange activity and thermostability, 1.43-fold activation by 1 mM (NH4)2SO4 (reaches plateau at 20 mM)

Lactococcus lactis

2.4.2.6

Rb+

enhances base exchange activity, 1.51-fold activation by 1 mM RbCl

Lactococcus lactis

2.4.2.6

17250

4 * 17250, elution as single peak from the preparative gel filtration column

Lactococcus lactis

2.4.2.6

18290

theoretical molecular weight

Lactococcus lactis

2.4.2.6

25500

according to migration in SDS-12.5% polyacrylamide gel electrophoresis

Lactococcus lactis

2.4.2.6

69000

gel filtration chromatography

Lactococcus lactis

2.4.2.6

Lactococcus lactis

preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed

?

2.4.2.6

purine base + 2-deoxy-D-ribosyl-base

Lactococcus lactis

purine deoxyribonucleoside + base

preferred over pyrimidine deoxyribonucleoside synthesis

r

2.4.2.6

pyrimidine base + 2-deoxy-D-ribosyl-base

Lactococcus lactis

pyrimidine deoxyribonucleoside + base

r

2.4.2.6

Lactococcus lactis

subsp. lactis JCM5808

2.4.2.6

preclearing of bacterial lysate with 40% ammonium sulfate and subsequent precipitation of proteins with 90% ammonium sulfate, gel filtration of dissolved protein on Sephacryl S-200 column, combination of fractions with N-deoxyribosyltransferase activity, precipitation of proteins with 90% ammonium sulfate, dialysis of dissolved proteins and concentration by ultrafiltration, second gel filtration on Sephacryl S-200 column, combination of active fractions and concentration by ultrafiltration, recovery of 70.4% of activity, storage at -20°C in 50% glycerol

Lactococcus lactis

2.4.2.6

0.9% in presence of 1 mM ZnCl2; 102% in presence of 1 mM NaCl; 105% in presence of 1 mM CsCl; 106% in presence of 1 mM NaSO4; 12.9% in presence of 1 mM CoCl2; 143% in presence of 1 mM NH4Cl; 151% in presence of 1 mM RbCl; 163% in presence of 1 mM (NH4)2SO4; 165% in presence of 1 mM K2SO4; 166% in presence of 1 mM KCl; 166% in presence of 1 mM (NH4)2SO4 and 1 mM KCl; 192% in presence of 10 mM KCl; 194% in presence of 10 mM (NH4)2SO4; 3.2% in presence of 1 mM CuSO4; 51.8% in presence of 1 mM CaCl2; 66.3% in presence of 1 mM MgCl2; 66% in presence of 1 mM MgSO4; 68.3% in presence of 1 mM FeCl2; 97.7% in presence of 1 mM LiCl

Lactococcus lactis

2.4.2.6

6.67

equals 100%, purified protein, in presence of 0.5 mg/ml bovine serum albumin, pH 7.5, 10 min, 25°C

Lactococcus lactis

2.4.2.6

adenine + deoxycytidine

64.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + cytosine

r

2.4.2.6

adenine + deoxyguanosine

46.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + guanine

r

2.4.2.6

adenine + deoxyinosine

57.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + hypoxanthine

r

2.4.2.6

adenine + deoxyuridine

75.4% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + uracil

r

2.4.2.6

adenine + thymidine

74.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + thymine

r

2.4.2.6

cytosine + deoxyadenosine

27.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + adenine

r

2.4.2.6

cytosine + deoxyguanosine

25.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + guanine

r

2.4.2.6

cytosine + deoxyinosine

32.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + hypoxanthine

r

2.4.2.6

cytosine + deoxyuridine

62.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + uracil

r

2.4.2.6

cytosine + thymidine

53.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + thymine

r

2.4.2.6

cytosine + thymidine

standard reaction for cofactor analyses as well as activating compound and inhibitor studies, in presence of 40 mM ammonium sulfate and 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + thymine

r

2.4.2.6

hypoxanthine + deoxyadenosine

38.0% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + adenine

r

2.4.2.6

hypoxanthine + deoxycytidine

51.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + cytosine

r

2.4.2.6

hypoxanthine + deoxyguanosine

25.2% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + guanine

r

2.4.2.6

hypoxanthine + deoxyuridine

65.6% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + uracil

r

2.4.2.6

hypoxanthine + thymidine

76.8% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + thymine

r

2.4.2.6

preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed

691053

Lactococcus lactis

?

2.4.2.6

ribonucleoside, arabinosylnucleoside and 2,3-dideoxyribonucleoside are no substrates for base-exchange by YejD

691053

Lactococcus lactis

?

2.4.2.6

purine base + 2-deoxy-D-ribosyl-base

691053

Lactococcus lactis

purine deoxyribonucleoside + base

preferred over pyrimidine deoxyribonucleoside synthesis

r

2.4.2.6

pyrimidine base + 2-deoxy-D-ribosyl-base

691053

Lactococcus lactis

pyrimidine deoxyribonucleoside + base

r

2.4.2.6

thymine + deoxyadenosine

22.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + adenine

r

2.4.2.6

thymine + deoxycytidine

38.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + cytosine

r

2.4.2.6

thymine + deoxyguanosine

17.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + guanine

r

2.4.2.6

thymine + deoxyinosine

19.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin, equilibrium reached after 2 h

691053

Lactococcus lactis

thymidine + hypoxanthine

r

2.4.2.6

thymine + deoxyuridine

53.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + uracil

r

2.4.2.6

uracil + deoxyadenosine

17.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + adenine

r

2.4.2.6

uracil + deoxycytidine

37.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + cytosine

r

2.4.2.6

uracil + deoxyguanosine

16.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + guanine

r

2.4.2.6

uracil + deoxyinosine

23.8% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + hypoxanthine

r

2.4.2.6

uracil + thymidine

43.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + thymine

r

2.4.2.6

homotetramer

4 * 17250, elution as single peak from the preparative gel filtration column

Lactococcus lactis

2.4.2.6

N-deoxyribosyltransferase

Lactococcus lactis

2.4.2.6

NTD

Lactococcus lactis

2.4.2.6

nucleoside N-deoxyribosyltransferase

Lactococcus lactis

2.4.2.6

trans-N-deoxyribosylase

Lactococcus lactis

2.4.2.6

YejD

Lactococcus lactis

2.4.2.6

30

assay at, pH 7.5

Lactococcus lactis

2.4.2.6

(NH4)2SO4 is more thermostabilizing than KCl

Lactococcus lactis

2.4.2.6

25

pH 7.5, 20 min, 60% activity lost, in presence of 10 mM (NH4)2SO4 only 20% activity lost, in presence of 500 mM KCL only 6% activity lost

Lactococcus lactis

2.4.2.6

30

pH 7.5, 20 min, in presence of 500 mM (NH4)2SO4 fully active

Lactococcus lactis

2.4.2.6

35

and more, pH 7.5, 20 min, 75% activity lost

Lactococcus lactis

2.4.2.6

7.5

assay at

Lactococcus lactis

2.4.2.6

K+

K+-activated type II enzyme: binding of K+ is required to enhance activity through induced conformational change

Lactococcus lactis

2.4.2.6

does not require phosphate derivatives such as ATP, thiamine diphosphate, pyridoxal phosphate to complete 2-deoxyribose transfer

Lactococcus lactis

2.4.2.6

in pET-22b(+) for expression in Escherichia coli BL21(DE3); in pGEM-T for cloning

Lactococcus lactis

2.4.2.6

Ca2+

inhibits base exchange activity at 1 mM

Lactococcus lactis

2.4.2.6

CO2

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

Cu2+

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

Mg2+

inhibits base exchange activity at 1mM

Lactococcus lactis

2.4.2.6

Mn2+

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

Zn2+

reduces base exchange activity by 87-99% at 1 mM

Lactococcus lactis

2.4.2.6

K+

enhances base exchange activity and thermostability, 1.66-fold activation by 1 mM KCl (reaches plateau at 20 mM)

Lactococcus lactis

2.4.2.6

enzymatic activity depends on monovalent cations as cofactors rather than catalysts; K+ and NH4+ have no synergistic effects; Li+ and Na+ do not activate

Lactococcus lactis

2.4.2.6

NH4+

enhances base exchange activity and thermostability, 1.43-fold activation by 1 mM (NH4)2SO4 (reaches plateau at 20 mM)

Lactococcus lactis

2.4.2.6

Rb+

enhances base exchange activity, 1.51-fold activation by 1 mM RbCl

Lactococcus lactis

2.4.2.6

17250

4 * 17250, elution as single peak from the preparative gel filtration column

Lactococcus lactis

2.4.2.6

18290

theoretical molecular weight

Lactococcus lactis

2.4.2.6

25500

according to migration in SDS-12.5% polyacrylamide gel electrophoresis

Lactococcus lactis

2.4.2.6

69000

gel filtration chromatography

Lactococcus lactis

2.4.2.6

Lactococcus lactis

preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed

?

2.4.2.6

purine base + 2-deoxy-D-ribosyl-base

Lactococcus lactis

purine deoxyribonucleoside + base

preferred over pyrimidine deoxyribonucleoside synthesis

r

2.4.2.6

pyrimidine base + 2-deoxy-D-ribosyl-base

Lactococcus lactis

pyrimidine deoxyribonucleoside + base

r

2.4.2.6

preclearing of bacterial lysate with 40% ammonium sulfate and subsequent precipitation of proteins with 90% ammonium sulfate, gel filtration of dissolved protein on Sephacryl S-200 column, combination of fractions with N-deoxyribosyltransferase activity, precipitation of proteins with 90% ammonium sulfate, dialysis of dissolved proteins and concentration by ultrafiltration, second gel filtration on Sephacryl S-200 column, combination of active fractions and concentration by ultrafiltration, recovery of 70.4% of activity, storage at -20°C in 50% glycerol

Lactococcus lactis

2.4.2.6

0.9% in presence of 1 mM ZnCl2; 102% in presence of 1 mM NaCl; 105% in presence of 1 mM CsCl; 106% in presence of 1 mM NaSO4; 12.9% in presence of 1 mM CoCl2; 143% in presence of 1 mM NH4Cl; 151% in presence of 1 mM RbCl; 163% in presence of 1 mM (NH4)2SO4; 165% in presence of 1 mM K2SO4; 166% in presence of 1 mM KCl; 166% in presence of 1 mM (NH4)2SO4 and 1 mM KCl; 192% in presence of 10 mM KCl; 194% in presence of 10 mM (NH4)2SO4; 3.2% in presence of 1 mM CuSO4; 51.8% in presence of 1 mM CaCl2; 66.3% in presence of 1 mM MgCl2; 66% in presence of 1 mM MgSO4; 68.3% in presence of 1 mM FeCl2; 97.7% in presence of 1 mM LiCl

Lactococcus lactis

2.4.2.6

6.67

equals 100%, purified protein, in presence of 0.5 mg/ml bovine serum albumin, pH 7.5, 10 min, 25°C

Lactococcus lactis

2.4.2.6

adenine + deoxycytidine

64.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + cytosine

r

2.4.2.6

adenine + deoxyguanosine

46.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + guanine

r

2.4.2.6

adenine + deoxyinosine

57.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + hypoxanthine

r

2.4.2.6

adenine + deoxyuridine

75.4% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + uracil

r

2.4.2.6

adenine + thymidine

74.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyadenosine + thymine

r

2.4.2.6

cytosine + deoxyadenosine

27.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + adenine

r

2.4.2.6

cytosine + deoxyguanosine

25.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + guanine

r

2.4.2.6

cytosine + deoxyinosine

32.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + hypoxanthine

r

2.4.2.6

cytosine + deoxyuridine

62.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + uracil

r

2.4.2.6

cytosine + thymidine

53.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + thymine

r

2.4.2.6

cytosine + thymidine

standard reaction for cofactor analyses as well as activating compound and inhibitor studies, in presence of 40 mM ammonium sulfate and 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxycytidine + thymine

r

2.4.2.6

hypoxanthine + deoxyadenosine

38.0% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + adenine

r

2.4.2.6

hypoxanthine + deoxycytidine

51.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + cytosine

r

2.4.2.6

hypoxanthine + deoxyguanosine

25.2% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + guanine

r

2.4.2.6

hypoxanthine + deoxyuridine

65.6% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + uracil

r

2.4.2.6

hypoxanthine + thymidine

76.8% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyinosine + thymine

r

2.4.2.6

preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed

691053

Lactococcus lactis

?

2.4.2.6

ribonucleoside, arabinosylnucleoside and 2,3-dideoxyribonucleoside are no substrates for base-exchange by YejD

691053

Lactococcus lactis

?

2.4.2.6

purine base + 2-deoxy-D-ribosyl-base

691053

Lactococcus lactis

purine deoxyribonucleoside + base

preferred over pyrimidine deoxyribonucleoside synthesis

r

2.4.2.6

pyrimidine base + 2-deoxy-D-ribosyl-base

691053

Lactococcus lactis

pyrimidine deoxyribonucleoside + base

r

2.4.2.6

thymine + deoxyadenosine

22.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + adenine

r

2.4.2.6

thymine + deoxycytidine

38.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + cytosine

r

2.4.2.6

thymine + deoxyguanosine

17.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + guanine

r

2.4.2.6

thymine + deoxyinosine

19.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin, equilibrium reached after 2 h

691053

Lactococcus lactis

thymidine + hypoxanthine

r

2.4.2.6

thymine + deoxyuridine

53.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

thymidine + uracil

r

2.4.2.6

uracil + deoxyadenosine

17.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + adenine

r

2.4.2.6

uracil + deoxycytidine

37.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + cytosine

r

2.4.2.6

uracil + deoxyguanosine

16.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + guanine

r

2.4.2.6

uracil + deoxyinosine

23.8% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + hypoxanthine

r

2.4.2.6

uracil + thymidine

43.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin

691053

Lactococcus lactis

deoxyuridine + thymine

r

2.4.2.6

homotetramer

4 * 17250, elution as single peak from the preparative gel filtration column

Lactococcus lactis

2.4.2.6

30

assay at, pH 7.5

Lactococcus lactis

2.4.2.6

(NH4)2SO4 is more thermostabilizing than KCl

Lactococcus lactis

2.4.2.6

25

pH 7.5, 20 min, 60% activity lost, in presence of 10 mM (NH4)2SO4 only 20% activity lost, in presence of 500 mM KCL only 6% activity lost

Lactococcus lactis

2.4.2.6

30

pH 7.5, 20 min, in presence of 500 mM (NH4)2SO4 fully active

Lactococcus lactis

2.4.2.6

35

and more, pH 7.5, 20 min, 75% activity lost

Lactococcus lactis

2.4.2.6

7.5

assay at

Lactococcus lactis