title: Microchip CE-LIF method for the hydrolysis of L-glutamine by using L-asparaginase enzyme reactor based on gold nanoparticle.
abstract: L-Asparaginase (L-Asnase) can suppress the growth of malignant cells by rapid depletion of two essential amino acids, L-glutamine (L-Gln) and L-asparagine (L-Asn). To study the cytotoxic effect and the secondary complications of L-Asnase in the treatment of acute lymphoblastic leukemia, the development of a novel enzyme reactor of L-Asnase for the hydrolysis of L-Gln, employing the enzyme-gold nanoparticle conjugates in capillary, was reported in this work. First, a microchip CE (MCE)-LIF was established for the separation of L-amino acids (L-Gln and L-glutamic acid) and studying the hydrolysis of L-Gln by using L-Asnase enzyme reactor. Then, using L-Gln as target analyte, the enzyme kinetics of L-Asnase in free solution, enzyme-gold nanoparticle conjugates (E-GNP), and the enzyme-gold nanoparticle conjugates immobilized in capillary (E-GNP-C) were investigated in detail with the proposed MCE-LIF method. Moreover, for optimizing the enzymatic reaction efficiency, three important parameters, including the length of capillary, the enzyme concentration reacted with gold nanoparticle and the amount of L-Asnase immobilized on the gold nanoparticle, have been studied. Owing to the high specific activity, the E-GNP-C enzyme reactor exhibited the best performance for the hydrolysis of L-Gln.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing, PR China.
authors.forename: Juan
authors.initials: J
authors.lastname: Qiao
authors.forename: Li
authors.initials: L
authors.lastname: Qi
authors.forename: Huijuan
authors.initials: H
authors.lastname: Yan
authors.forename: Yaping
authors.initials: Y
authors.lastname: Li
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Mu
mesh_terms: D001215:Asparaginase / Q000737:chemistry / Q000378:metabolism*; D019149:Bioreactors; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D018698:Glutamic Acid / Q000302:isolation & purification; D005973:Glutamine / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D006046:Gold / Q000737:chemistry; D006868:Hydrolysis; D057230:Limit of Detection; D016014:Linear Models; D053768:Metal Nanoparticles / Q000737:chemistry*; D015203:Reproducibility of Results
chemical_list: D004800:Enzymes, Immobilized; D005973:Glutamine; D018698:Glutamic Acid; D006046:Gold; D001215:Asparaginase