A Process to Produce Penicillin G Acylase by Surface-Adhesion Fermentation Using <Emphasis Type="Italic">Mucor griseocyanus</Emphasis> to Obtain 6-Aminopenicillanic Acid by Penicillin G Hydrolysis |
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Authors: | José Luis Martínez-Hernández Marco Arnulfo Mata-Gómez Cristóbal Noé Aguilar-González Anna Ilyina |
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Institution: | 1.Department of Biotechnology,Chemistry School, Universidad Autónoma de Coahuila,Saltillo,Mexico;2.Department of Food Research,School of Chemistry, Universidad Autónoma de Coahuila,Saltillo,Mexico |
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Abstract: | The production of extracellular and mycelia-associated penicillin G acylase (maPGA) with Mucor griseocyanus H/55.1.1 by surface-adhesion fermentation using Opuntia imbricata, a cactus, as a natural immobilization support was studied. Enzyme activity to form 6-aminopencillanic acid (6-APA) from
penicillin G was assayed spectrophotometrically. The penicillin G hydrolysis to 6-APA was evaluated at six different times
using PGA samples recovered from the skim milk medium at five different incubation times. Additionally, the effect of varying
the penicillin G substrate concentration level on the PGA enzyme activity was also studied. The maximum reaction rate, V
max, and the Michaelis constant, K
M, were determined using the Michaelis–Menten model. The maximum levels for maPGA and extracellular activity were found to
be 2,126.50 international unit per liter (IU/l; equal to 997.83 IU/g of support) at 48 h and 755.33 IU/l at 60 h, respectively.
Kinetics of biomass production for total biomass showed a maximum growth at 60 h of 3.36 and 2.55 g/l (equal to 0.012 g of
biomass per gram of support) for the immobilized M. griseocyanus biomass. The maPGA was employed for the hydrolysis of penicillin G to obtain 6-APA in a batch reactor. The highest quantity
of 6-APA obtained was 226.16 mg/l after 40-min reaction. The effect of substrate concentration on maPGA activity was evaluated
at different concentrations of penicillin G (0–10 mM). K
M and V
max were determined to be 3.0 × 10−3 M and 4.4 × 10−3 mM/min, respectively. |
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