Characterization of the penicillin G acylase from Bacillus megaterium ATCC 14945

Souza, Vanessa Ribeiro de; Silva, Ana C. G.; Pinotti, L.; Araújo, Heloísa Sobreiro Selistre de; Giordano, Raquel de Lima Camargo

doi:10.1590/s1516-89132005000400013

Cited by 25 publications

(13 citation statements)

References 11 publications

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“…The PGA from Escherichia coli is an intracellular enzyme located in the periplasmic space, consisting of two different sub-units, ␣ and ␤, with molecular weights of 20,5 and 69 kDa, respectively, with a pI between 6.7 and 6.9 [4][5][6]. The enzyme produced by B. megaterium is extracellar and is also constituted of two sub-units, with average molecular mass of 26.9 and 59.1 kDa [7].…”

Section: Introductionmentioning

confidence: 99%

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Pinotti

Fonseca

Prazeres

et al. 2009

Biochemical Engineering Journal

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Section: Introductionmentioning

confidence: 99%

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Pinotti

Fonseca

Prazeres

et al. 2009

Biochemical Engineering Journal

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“…This bacterium secretes the enzyme to the medium (one important aspect in this process), thus reducing downstream costs. Results concerning medium optimization for this process may be found elsewhere [7][8][9][10][11][12][13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Monitoring bioreactors using principal component analysis: production of penicillin G acylase as a case study

Nucci

Cruz

Giordano

2009

Bioprocess Biosyst Eng

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The complexity of biological processes often makes impractical the development of detailed, structured phenomenological models of the cultivation of microorganisms in bioreactors. In this context, data pre-treatment techniques are useful for bioprocess control and fault detection. Among them, principal component analysis (PCA) plays an important role. This work presents a case study of the application of this technique during real experiments, where the enzyme penicillin G acylase (PGA) was produced by Bacillus megaterium ATCC 14945. PGA hydrolyzes penicillin G to yield 6-aminopenicilanic acid (6-APA) and phenyl acetic acid. 6-APA is used to produce semi-synthetic beta-lactam antibiotics. A static PCA algorithm was implemented for on-line detection of deviations from the desired process behavior. The experiments were carried out in a 2-L bioreactor. Hotteling's T(2) was the discrimination criterion employed in this multivariable problem and the method showed a high sensibility for fault detection in all real cases that were studied.

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“…3.5.1.11) is a critical enzyme that is extensively used by the pharmaceutical industry in the hydrolysis reaction of penicillin G to produce two important intermediates: phenylacetic acid and 6-APA. The latter that is of most interest is an intermediate for the production of semisynthetic penicillins, such as amoxycillin and ampicillin [3], and is mainly produced by enzymatic or chemical deacylation [4] of the natural benzyl penicillin. Penicillin acylases or amidases are used in soluble or immobilized form under acidic or alkaline conditions for the enzymatic conversion of penicillins to 6-APA and semisynthetic penicillins [3].…”

Section: Introductionmentioning

confidence: 99%

A Process to Produce Penicillin G Acylase by Surface-Adhesion Fermentation Using Mucor griseocyanus to Obtain 6-Aminopenicillanic Acid by Penicillin G Hydrolysis

Martínez-Hernández

Mata-Gómez

Aguilar-González

et al. 2009

Appl Biochem Biotechnol

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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 x 10(-3) M and 4.4 x 10(-3) mM/min, respectively.

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Characterization of the penicillin G acylase from Bacillus megaterium ATCC 14945

Abstract: The purpose of this work was to characterize the enzyme penicillin G acylase (PGA)

Cited by 25 publications

References 11 publications

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Monitoring bioreactors using principal component analysis: production of penicillin G acylase as a case study

A Process to Produce Penicillin G Acylase by Surface-Adhesion Fermentation Using Mucor griseocyanus to Obtain 6-Aminopenicillanic Acid by Penicillin G Hydrolysis

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