Table 2.

Screening reactions and detection strategies

Enzyme name (EC number)	Chemical reactions	Detection strategy	Control wells*
Adenylosuccinate synthetase (6.3.4.4)	Primary: GTP + IMP + L-aspartate → GDP + Pi + adenylosuccinate	Detect GDP using Cisbio Transcreener ADP kit.	Hadacidin [42]
Choline kinase (2.7.1.32)	Primary: ATP + choline → ADP + phosphocholine	Detect ADP using Cisbio Transcreener ADP kit.	No choline
dUTPase (3.6.1.23)	Primary: dUTP + H2O → dUMP + PPi	Detect PPi with Lonza PPiLight kit.	CAS: 14270-73-6,84472-83-3
Farnesyl pyrophosphate synthase (2.5.1.−)	Primary: geranyl diphosphate + isopentenyl diphosphate → farnesyl diphosphate + PPi	Detect PPi with Lonza PPiLight kit.	compound 11 from [67]
Glutamate dehydrogenase (1.4.1.4)	Primary: L-glutamate + H2O + NADP+ → 2-oxoglutarate + NH3 + NADPH + H+	Detect NADPH fluorescence.	No glutamate
Guanylate kinase (2.7.4.8)	Primary: ATP + GMP → ADP + GDP	Detect ADP and GDP using Cisbio Transcreener ADP kit.	Iodoacetamide
N-myristoyltransferase (2.3.1.97)	Primary: myristoyl-CoA + glycylpeptide → CoA + N-myristoylglycylpeptide	Detect CoA’s free –SH groups with ThioGlo.	aurintricarboxylate
OMP decarboxylase (4.1.1.23)	Primary: OMP → UMP + ATP CMP kinase: UMP + ATP → UDP + ADP	Detect ADP using Cisbio Transcreener ADP kit.	No OMP
S- adenosylhomocysteine hydrolase (3.3.1.1)	Primary: S-adenosylhomocysteine + H2O → adenosine + homocysteine Adenosine deaminase: adenosine → inosine + NH3	Detect homocysteine’s free –SH groups with ThioGlo.	No S- adenosylhomocysteine

^*Control wells either contained a known inhibitor of the enzyme or lacked a substrate so as to mimic the condition of complete inhibition of the enzyme.