Biocalorimetric and respirometric studies on production of Penicillin G acylase from Bacillus badius pac in E. coli DH5α

Karthikeyan, Raman; Surianarayanan, M.; Sekar, Sudharshan; Gunasekaran, Paramasamy; Baran, Mandal Asit

doi:10.1016/j.bej.2011.05.003

Cited by 15 publications

(5 citation statements)

References 26 publications

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“…In our studies we obtained values from 335 to 460 kJ mol −1 (Table 1) for different aeration and agitation rates. The deviation from the theoretical values was reported33 to be due to the degree of aerobicity, partly anaerobic nature of the organism and the inability to precisely measure the low OUR occurring at low values of substrate reductions. In our studies at optimized conditions, Y Q/O value was 460 kJ mol −1 .…”

Section: Resultsmentioning

confidence: 90%

Bioenergetics and pathway of acid blue 113 degradation byStaphylococcus lentus

Sekar

Surianarayanan

Shanmugam

et al. 2012

Biotechnology Progress

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Bioreaction calorimetric studies of degradation of the dye acid blue 113 by Staphylococcus lentus are reported for the first time. The heat released during the dye degradation process can be successfully measured using reaction calorimeter. Power time and oxygen uptake rate (OUR) profile followed each other suggesting that heat profiles could monitor the progress of the dye degradation in biocalorimetry. The shifts observed in power-time profile indicated three distinct phases of the bioprocess indicating simultaneous utilization of glucose (primary) and dye (secondary carbon source). Secretion of azoreductase enzyme enhanced the degradation process. Optimization of aeration and agitation rates was observed to be vital to efficient dye degradation. The degradative pathway for acid blue 113 by S. lentus was delineated via high-performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FT-IR), and gas chromatography coupled with mass spectrometry (GC-MS) analyses. Interestingly the products of degradation were found to have low toxicity, as per cytotoxicity measurements.

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Section: Resultsmentioning

confidence: 90%

Bioenergetics and pathway of acid blue 113 degradation byStaphylococcus lentus

Sekar

Surianarayanan

Shanmugam

et al. 2012

Biotechnology Progress

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“…BmPGA, a PGA that is extracellularly expressed at high levels in Bacillus subtilis WB600, is currently used in the hydrolysis of penicillin G and cephalosporin G to produce nucleus 6-APA and 7-ADCA ( Illanes et al, 1994 ; Yang et al, 2001 ). Besides, BbPGA is also proven to be capable of efficiently converting penicillin G to 6-APA ( Karthikeyan et al, 2011 ). By contrast, AvPGA has a higher S/H ratio than EcPGA (13 vs. 9.6) in the synthesis of cephalosporin C, indicating a good enzyme source with high synthetic activity ( Terreni et al, 2007 ).…”

Section: Improving the Enzymatic Propertymentioning

confidence: 99%

“…The PGAs associated with antibiotic production include the periplasmic enzymes from Gram-negative bacteria, such as Escherichia coli (EcPGA), Kluyvera cryocrescens (KcPGA), Providencia rettgeri (PrPGA), Achromobacter xylosoxidans (AxPGA), Alcaligenes faecalis (AfPGA), and the extracellular enzymes from Gram-positive bacteria, such as Bacillus megaterium (BmPGA), Arthrobacter viscosus (AvPGA) (Figure 3) (Grulich et al, 2013;Srirangan et al, 2013). Intriguingly, one PGA extracted from Gram-positive bacteria Bacillus badius (BbPGA) is an intracellular enzyme (Karthikeyan et al, 2011). Due to the wide variation in the intrinsic properties of different PGAs, their synthetic activities, especially the S/H ratios, are significantly different (Table 1).…”

Section: Mining Of Enzymesmentioning

confidence: 99%

Strategies to Improve the Biosynthesis of β-Lactam Antibiotics by Penicillin G Acylase: Progress and Prospects

Pan

et al. 2022

Front. Bioeng. Biotechnol.

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β-Lactam antibiotics are widely used anti-infection drugs that are traditionally synthesized via a chemical process. In recent years, with the growing demand for green alternatives, scientists have turned to enzymatic synthesis. Penicillin G acylase (PGA) is the second most commercially used enzyme worldwide with both hydrolytic and synthetic activities toward antibiotics, which has been used to manufacture the key antibiotic nucleus on an industrial level. However, the large-scale application of PGA-catalyzed antibiotics biosynthesis is still in the experimental stage because of some key limitations, such as low substrate concentration, unsatisfactory yield, and lack of superior biocatalysts. This paper systematically reviews the strategies adopted to improve the biosynthesis of β-lactam antibiotics by adjusting the enzymatic property and manipulating the reaction system in recent 20 years, including mining of enzymes, protein engineering, solvent engineering, in situ product removal, and one-pot reaction cascade. These advances will provide important guidelines for the future use of enzymatic synthesis in the industrial production of β-lactam antibiotics.

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“…The production of PGA in bioreactor cultivations has been widely studied using both batch [ 20 - 22 ] and high cell density fed-batch cultures [ 6 , 23 , 24 ]. Fed-batch cultivation is considered to be the most efficient process for the production of inducible heterologous proteins, including PGA, because the nutrient supply can be modulated to firstly achieve high biomass formation and then proceed with the induction [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

High-throughput strategies for penicillin G acylase production in rE. colifed-batch cultivations

et al. 2014

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BackgroundPenicillin G acylase (PGA) is used industrially to catalyze the hydrolysis of penicillin G to obtain 6-aminopenicillanic acid. In Escherichia coli, the most-studied microorganism for PGA production, this enzyme accumulates in the periplasmic cell space, and temperature plays an important role in the correct synthesis of its subunits.ResultsThis work investigates the influence of medium composition, cultivation strategy, and temperature on PGA production by recombinant E. coli cells. Shake flask cultures carried out using induction temperatures ranging from 18 to 28°C revealed that the specific enzyme activity achieved at 20°C (3000 IU gDCW-1) was 6-fold higher than the value obtained at 28°C. Auto-induction and high cell density fed-batch bioreactor cultures were performed using the selected induction temperature, with both defined and complex media, and IPTG and lactose as inducers. Final biomass concentrations of 100 and 120 gDCW L-1, and maximum enzyme productivities of 7800 and 5556 IU L-1 h-1, were achieved for high cell density cultures using complex and defined media, respectively.ConclusionsTo the best of our knowledge, the volumetric enzyme activity and productivity values achieved using the complex medium are the highest ever reported for PGA production using E. coli. Overall PGA recovery yields of 64 and 72% after purification were achieved for crude extracts obtained from cells cultivated in defined and complex media, respectively. The complex medium was the most cost-effective for PGA production, and could be used in both high cell density and straightforward auto-induction protocols.

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Biocalorimetric and respirometric studies on production of Penicillin G acylase from Bacillus badius pac in E. coli DH5α

Cited by 15 publications

References 26 publications

Bioenergetics and pathway of acid blue 113 degradation byStaphylococcus lentus

Bioenergetics and pathway of acid blue 113 degradation byStaphylococcus lentus

Strategies to Improve the Biosynthesis of β-Lactam Antibiotics by Penicillin G Acylase: Progress and Prospects

High-throughput strategies for penicillin G acylase production in rE. colifed-batch cultivations

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