TY - JOUR
T1 - Poly-lysine supported cross-linked enzyme aggregates of penicillin G acylase and its application in synthesis of β-lactam antibiotics
AU - Ye, Jiajie
AU - Li, Anni
AU - Chu, Tianshu
AU - Pan, Xin
AU - He, Bingfang
N1 - Publisher Copyright:
© 2019 Elsevier B.V.
PY - 2019/11/1
Y1 - 2019/11/1
N2 - Penicillin G acylase (PGA) from Providencia rettgeri PX04 (PrPGA) was utilized to synthesize β-lactam antibiotics. Poly-lysine supported cross-linked enzyme aggregates (PL-CLEAs) were prepared using PGA. Addition of poly-lysine significantly increased retention of PGA activity in CLEAs, with a decrease in the synthesis/hydrolysis (S/H) ratio. PL-CLEAs with 0.56 mg/mL poly-lysine retained 83% of free PGA activity, and displayed a higher S/H ratio than that of the free enzyme. Both PL-CLEAs and CLEAs exhibited high pH and thermal stabilities. PL-CLEAs possessed the best stability profile, and the lowest α value [(kcat/Km)Ps/(kcat/Km)AD], and was most effective at amoxicillin synthesis. A >94% yield of amoxicillin was achieved using a D-HPGME/6-APA ratio of 1.2:1 (240 mM, 200 mM), with fed-batch addition of D-HPGME. PL-CLEAs displayed excellent operational stability during amoxicillin synthesis. Over 97% of initial conversion was retained after twenty rounds of catalysis. PL-CLEAs exhibited greater potency than CLEAs in practical catalysis, permitting a higher concentration of reactants.
AB - Penicillin G acylase (PGA) from Providencia rettgeri PX04 (PrPGA) was utilized to synthesize β-lactam antibiotics. Poly-lysine supported cross-linked enzyme aggregates (PL-CLEAs) were prepared using PGA. Addition of poly-lysine significantly increased retention of PGA activity in CLEAs, with a decrease in the synthesis/hydrolysis (S/H) ratio. PL-CLEAs with 0.56 mg/mL poly-lysine retained 83% of free PGA activity, and displayed a higher S/H ratio than that of the free enzyme. Both PL-CLEAs and CLEAs exhibited high pH and thermal stabilities. PL-CLEAs possessed the best stability profile, and the lowest α value [(kcat/Km)Ps/(kcat/Km)AD], and was most effective at amoxicillin synthesis. A >94% yield of amoxicillin was achieved using a D-HPGME/6-APA ratio of 1.2:1 (240 mM, 200 mM), with fed-batch addition of D-HPGME. PL-CLEAs displayed excellent operational stability during amoxicillin synthesis. Over 97% of initial conversion was retained after twenty rounds of catalysis. PL-CLEAs exhibited greater potency than CLEAs in practical catalysis, permitting a higher concentration of reactants.
KW - Antibiotics
KW - CLEAs
KW - Kinetically controlled synthesis
KW - Penicillin G acylase
KW - Poly-lysine
UR - http://www.scopus.com/inward/record.url?scp=85070997398&partnerID=8YFLogxK
U2 - 10.1016/j.ijbiomac.2019.08.021
DO - 10.1016/j.ijbiomac.2019.08.021
M3 - 文章
C2 - 31381925
AN - SCOPUS:85070997398
SN - 0141-8130
VL - 140
SP - 423
EP - 428
JO - International Journal of Biological Macromolecules
JF - International Journal of Biological Macromolecules
ER -