Mono sodium hydrogen phosphate

Bovine insulin, citrate synthase (CS) from porcine heart, Dithiothreitol (DTT) and 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid, dipotassium salt (bis-ANS) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Alexa fluor 350 and Alexa fluor 488 protein labeling kits (Catalog Nos. A-10170 and A-10235, respectively) were obtained from Molecular Probes (Invitrogen, Carlsbad, CA, USA). Malate dehydrogenase (MDH) from porcine heart, disodium hydrogen phosphate and mono sodium hydrogen phosphate were from Sisco Research Laboratories, India. All other chemicals were of analytical grade.

The commercial l-asparaginase
II, Bionase 5000 IU (Zydus
, India) available as lyophilized powder, was used throughout the
study. Monosodium hydrogen phosphate, disodium hydrogen phosphate,
glycerol (99.5%, molecular biology grade), and boric acid were purchased
from Sisco Research Laboratories Pvt., Ltd. (Mumbai, India). l-Asn, d-Gly, d-Asp, l-Asp, and mushroom
tyrosinase K were purchased from Sigma Aldrich (St. Louis, MO, USA).
The peptide corresponding to the lid loop GGTIAGGGDSATKSNYTAGKVG,
used in the study, was purchased from Creative Biolabs (Shirley, New
York, United States). All buffers were prepared fresh and used within
2 weeks of preparation. The plasmid pET28a encoding the wild-type
sequence of N term 6X-His-tagged l-asparaginase was obtained
from Genscript (USA).