Direct determination of the cephalosporin transforming activity of immobilized cells with use of an enzyme thermistor. 1. Verification of the mathematical model

Gemeiner, Peter; Štefuca, Vladimı́r; Welwardová, A.; Michalková, Eva; Welward, L.; Kurillová, L'.; Danielsson, Bengt

doi:10.1016/0141-0229(93)90115-i

Cited by 35 publications

(8 citation statements)

References 8 publications

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“…Oxidative deamination of d ‐amino acids to give α‐keto acids, ammonia, and hydrogen peroxide has lead to increasing biotechnological interest in the production of pure l ‐amino acids from racemic mixtures (Nakajima et al, 1990), the production of α‐keto acids with potential pharmacological activity (Brodelius et al, 1981; Butó et al, 1994; Trost and Fischer, 2002), the production of biosensors (Gemeiner et al, 1993), and most importantly, the industrial bioconversion of cephalosporin C to glutaryl‐7‐amino cephalosporanic acid, which is subsequently transformed enzymatically by glutaryl‐7‐ACA acylase into 7‐amino cephalosporanic acid (7‐ACA), a starting compound for the production of semisynthetic β‐lactam compounds (Szwajcer‐Dey et al, 1991; Conlon et al, 1995; Pilone et al, 1995; Sánchez‐Ferrer et al, 2004). All of the above biotransformations have been carried out to date by d ‐amino acid oxidase (EC 1.4.3.3, DAAO), but its recombinant production is difficult in procariotic organisms such as E. coli and its operational stability from the industrial point of view remains low (∼50 cycles) (Lin et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Maximization of Production of His-Tagged Glycine Oxidase and Its M261 Mutant Proteins

et al. 2006

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Glycine oxidase (GOX) from Bacillus subtilis is a new flavoprotein of great potential biotechnological use that catalizes the oxidative deamination of various amines (glycine, sarcosine, and N-ethyl-glycine) and D-amino acids (D-alanine and D-proline). However, its commercial application is hindered by its low heterologous expression in Escherichia coli due to its codon bias and the sensitivity of its N-terminus to proteases. The first problem has been solved by cloning the GOX gene from B. subtilis ATCC 6633 into the Rosetta E. coli strain, which contains the pRARE plasmid. The second problem was overcome by inserting the gene in the pET28a expression vector, which not only has a 6 x His tag but also increases the N-terminus in 36 amino acids without impairing either the enzymatic activity or the ribosome binding region. After induction with 0.5 mM isopropyl thio-beta-D-galactoside for 5 h in TB-medium, the soluble and active chimeric GOX was expressed up to 15.81 U x g(-1) cell, with a fermentation yield of 399 U x L(-1). The latter value represents about 16% of the total soluble protein content of the cell. The three latter values are higher than the best found in the literature by 16-, 28- and 4-fold, respectively. The enzyme was purified with a nickel HiTrap chelating-affinity column in 96% yield to apparent homogeneity. It was fully active and was stable for months at -80 degrees C in the presence of 10% glycerol. Its substrate specificity was similar to that previously described, but the constructed M261 mutants unexpectedly decreased in K(M) compared with the wild-type, especially in the M261Y mutant. Noteworthy, there was decrease in the K(M) for N-ethyl-glycine of up to 0.7 mM, similar to that found with N-alkyl-glycine oxidase. Such mutants open up new possible uses of this enzyme not only in the pharmacological industry but also in the clinical field for diabetic complications.

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Section: Introductionmentioning

confidence: 99%

Maximization of Production of His-Tagged Glycine Oxidase and Its M261 Mutant Proteins

et al. 2006

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“…The D-AOs are highly enantioselective enzymes and have a very broad substrate spectrum. This makes them biocatalysts that are in demand in several areas of biochemistry and biotechnology; for instance, they are used in qualitative and quantitative analyses of D-amino acids (13), in biosensors (12,21), in the production of L-amino acids (10,22), in the production of ␣-keto acids (5), and, most importantly, in the conversion of cephalosporin C to 7-glutarylcephalosporanic acid (26,35). The latter process takes place on an industrial scale, and the D-AO from T. variabilis is used as one of the two biocatalysts that take part in the enzymatic two-step conversion of cephalosporin C to 7-aminocephalosporanic acid (7), which is a key compound for the production of many semisynthetic ␤-lactam drugs (world market, $20.5 ϫ 10 9 ).…”

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confidence: 99%

Induction of the d-Amino Acid Oxidase from Trigonopsis variabilis

Hörner¹,

Wagner²,

Fischer³

1996

Appl Environ Microbiol

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Induction of the D-amino acid oxidase (EC. 1.4.3.3) from the yeast Trigonopsis variabilis was investigated by using a minimal medium containing glucose as the carbon and energy source, (NH 4) 2 SO 4 as the nitrogen source, and various D-and DL-amino acid derivatives as inducers. The best new inducers found were Ncarbamoyl-D-alanine, N-acetyl-D-tryptophan, and N-chloroacetyl-D-␣-aminobutyric acid; when the induction effects of these compounds were compared with the effects of D-alanine as the nitrogen source and inducer, the resulting activities of D-amino acid oxidase per gram of dried yeast were 4.2, 2.1, and 1.5 times higher, respectively. The optimum concentration of the best inducer, N-carbamoyl-D-alanine, was 5 mM. This inducer could also be used in its racemic form. The induction was pH dependent. After cultivation of the yeast in a 50-liter bioreactor, D-amino acid oxidase activity of about 3,850 kat (231,000 U) was obtained. In addition, production of the D-amino acid oxidase was found to be significantly dependent on the metal salt composition of the medium. Addition of zinc ions was required to obtain high D-amino acid oxidase levels in the cells. The optimum concentration of ZnSO 4 was about 140 M.

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“…Similarly, the results of this technique were useful in the selection of Trigonopsis variabilis strains for high cephalosporin-transforming activity [60]. Also, the cephalosporin-transforming activity of D-amino acid oxidase isolated from yeast was identified in a similar manner.…”

Section: Miscellaneous Applicationsmentioning

confidence: 81%

Principles of Enzyme Thermistor Systems: Applications to Biomedical and Other Measurements

Xie

Ramanathan

Danielsson

1999

Advances in Biochemical Engineering/Biotechnology

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This chapter presents an overview of thermistor-based calorimetric measurements. Bioanalytical applications are emphasized from both the chemical and biomedical points of view. The introductory section elucidates the principles involved in the thermometric measurements. The following section describes in detail the evolution of the various versions of enzyme-thermistor devices. Special emphasis is laid on the description of modern "mini" and "miniaturized" versions of enzyme thermistors. Hybrid devices are also introduced in this section. In the sections on applications, the clinical/biomedical areas are dealt with separately, followed by other applications. Mention is also made of miscellaneous applications. A special section is devoted to future developments, wherein novel concepts of telemedicine and home diagnostics are highlighted. The role of communication and information technology in telemedicine is also mentioned. In the concluding sections, an attempt is made to incorporate the most recent references on specific topics based on enzyme-thermistor systems.

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Direct determination of the cephalosporin transforming activity of immobilized cells with use of an enzyme thermistor. 1. Verification of the mathematical model

Cited by 35 publications

References 8 publications

Maximization of Production of His-Tagged Glycine Oxidase and Its M261 Mutant Proteins

Maximization of Production of His-Tagged Glycine Oxidase and Its M261 Mutant Proteins

Induction of the d-Amino Acid Oxidase from Trigonopsis variabilis

Principles of Enzyme Thermistor Systems: Applications to Biomedical and Other Measurements

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