Activity and stability of immobilized penicillin amidase at low pH values

Ferreira, Juliana de Souza; Straathof, Adrie J. J.; Franco, Telma Teixeira; Wielen, Luuk A.M. van der

doi:10.1016/j.molcatb.2003.09.003

Cited by 26 publications

(24 citation statements)

References 34 publications

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“…The literature reveals a half-life of 12 min at pH 3 and 378C for the native penicillin acylase opposed to 95 h at pH 8 and 378C (Masarova et al, 2001). Immobilization of a commercial penicillin acylase was reported improve stability when the pH drops from 8 to 3 even though the enzyme activity decreased by 80% (Ferreira et al, 2003). This confirms the possibility of using biphasic systems under extremely acidic conditions for the penG hydrolysis.…”

Section: Introductionmentioning

confidence: 56%

Novel reactive perstraction system applied to the hydrolysis of penicillin G

Wyss

Seitert

Stockar

et al. 2005

Biotech & Bioengineering

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The activity of penicillin acylase has been studied in aqueous and organic solvents, as free enzyme as well as immobilized within the membrane of liquid-core capsules. The activity of the enzyme is inhibited by the accumulation of the products of the hydrolysis reaction, namely phenyl acetic acid (PAA). In order to overcome this inhibition a range of organic solvents were tested for use in in situ product recovery. Of these solvents dibutyl sebacate (DBS) was chosen due to the rapid extraction rate, the high logP and to facilitate capsule production. The extraction efficiency at pH 3.5 for PAA was >80% for phase ratios of >50% free solvent with partition coefficients of 8 and 0.7 for PAA and penicillin G (PenG), respectively, thereby showing that PAA could be selectively extracted at pH 3.5 and 25 degrees C. Liquid-core capsules containing DBS were shown to efficiently remove PAA selectively and the PAA could be effectively back-extracted and the capsules re-used in a three-stage process resulting in high product separation. Immobilization of penicillin acylase onto the capsule membranes resulted in increased operational stability of the enzyme and a very high enzyme activity. Over 53.3% of the PAA formed could be recovered in the capsule core with a concentration over sevenfold higher than in the aqueous phase. Higher extraction efficiencies could be obtained by varying the substrate concentration and number of capsules. The enzyme immobilized on capsules could be stored for over 4 months at pH 8 and 4 degrees C with no loss of activity. Over 80% of the initial activity could be recovered over five repeated batch cycles of the bioconversion process. The importance of capsular perstraction and reactive capsular perstraction has been clearly demonstrated.

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Section: Introductionmentioning

confidence: 56%

Novel reactive perstraction system applied to the hydrolysis of penicillin G

Wyss

Seitert

Stockar

et al. 2005

Biotech & Bioengineering

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“…In contrast, the equilibrium pH of the enzymatic hydrolysis in the cloud point system in every case was significantly higher than that of in the aqueous solution. The extraction of PAA from the diluted phase into the coacervate phase of cloud point system makes the diluted phase not be further acidified [5] and therefore maintains a relatively high pH till the enzymatic hydrolysis reaction approaches to its hydrolysis equilibrium. The influence of some key operation parameters in cloud point system, such as initial pH, substrate concentration, surfactant and immobilized penicillin acylase loads, was examined by adjusting one parameter at a time and keeping the other parameters at the fixed values respectively.…”

Section: Resultsmentioning

confidence: 99%

“…The enzymatic hydrolysis process is controlled by its thermodynamic equilibrium under the condition of without pH control [23]. As the PAA dissociation constant (pK) is about 4.3 and the aqueous-organic solvent extraction only works for a non-charged organic substance, the extractive enzymatic hydrolysis in an aqueous-organic solvent biphasic system restricts its effectiveness to a relatively low pH range [5]. Different from the aqueousorganic solvent biphasic system, the extraction of organic substance with charged ions in cloud point system has been reported [15].…”

Section: Resultsmentioning

confidence: 99%

“…It is known as 'Green Chemistry' or 'Sustainable Technology'. Extractive biocatalysis, i.e., coupling of a bioreaction unit with a product extraction unit in a biphasic system, has been described for enzymatic hydrolysis of Pen G by many researchers [2][3][4][5][6][7]. The extraction of phenylacetic acid (PAA) to a non-biocatalysis reaction phase makes the enzymatic hydrolysis without pH control possible [2,3], even direct hydrolysis of the Pen G from its fermentation broth [4].…”

Section: Introductionmentioning

confidence: 99%

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An Eco-Friendly and Sustainable Process for Enzymatic Hydrolysis of Penicillin G in Cloud Point System

Wang

et al. 2006

Bioprocess Biosyst Eng

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Enzymatic hydrolysis of penicillin G by immobilized penicillin acylase in a nonionic surfactant mediated cloud point system was presented. The effect of the operation parameters on equilibrium pH of this enzymatic hydrolysis process without pH control was examined. A relatively high equilibrium pH in cloud point system without pH control can be obtained. The feasibility of recycling utilization of the nonionic surfactant, a novel green solvent, was also investigated experimentally. Enzymatic hydrolysis of penicillin G in a discrete semi-batch mode, which simulates a semi-continuous process, envisages a completely eco-friendly, sustainable and efficient process for production of 6-aminopenicillanic acid.

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“…Availability of the immobilized enzyme catalysts with improved activity and stability is expected to reduce the expense of producing 6-APA and semisynthetic penicillins [2]. To obtain the desired immobilization and catalysis, various new carriers and technologies have been used to immobilize PA [3][4][5][6] and PA preparations were used to hydrolyze penicillin G in non-conventional medium [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Penicillin acylase immobilization depending on macromolecular crowding and catalysis in aqueous–organic medium

Xue

Wang

Zhou

et al. 2009

Bioprocess Biosyst Eng

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To enhance penicillin acylase (PA) performance, it was immobilized in mesocellular silica foams (MCFs) depended on macromolecular crowding and applied to catalysis in non-aqueous medium. Ficoll 70, dextran 10,000, dextran 40,000 and bovine serum albumin were co-assembled with PA. It was observed that specific activity of PA assembled in MCFs with dextran 10,000 in 80% cyclohexane (v/v) was 233.2 U/mg, 200% as that of PA assembled in MCFs in 80% cyclohexane and 323.5% as that of free PA in full aqueous medium. As content of alkane increased, activity of PA in MCFs with macromolecules varied slightly. In addition, PA co-immobilized with dextran 10 in MCFs retained 58.2% of its initial activity after heating at 50 degrees C for 4 h, 1.2 times higher than that of PA immobilized alone in MCFs. The results showed that macromolecular crowding was favorable for immobilization of PA and its catalysis in suitable aqueous-organic medium.

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Activity and stability of immobilized penicillin amidase at low pH values

Cited by 26 publications

References 34 publications

Novel reactive perstraction system applied to the hydrolysis of penicillin G

Novel reactive perstraction system applied to the hydrolysis of penicillin G

An Eco-Friendly and Sustainable Process for Enzymatic Hydrolysis of Penicillin G in Cloud Point System

Penicillin acylase immobilization depending on macromolecular crowding and catalysis in aqueous–organic medium

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