Optimization of culture condition for the production of D-amino acid oxidase in a recombinant Escherichia coli

2008 ◽  
Vol 13 (2) ◽  
pp. 144-149 ◽  
Author(s):  
Sae-Jin Kim ◽  
Nag-Jong Kim ◽  
Chang-Hun Shin ◽  
Chan-Wha Kim
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Culture Condition ◽  
Recombinant Escherichia Coli ◽  
Acid Oxidase ◽  
Amino Acid Oxidase

Bioprocess development of the production of the mutant P-219-L human d-amino acid oxidase for high soluble fraction expression in recombinant Escherichia coli

2010 ◽  
Vol 52 (2-3) ◽  
pp. 236-247 ◽  
Author(s):  
Rabab M. Abou El-Magd ◽  
Chizuru Sasaki ◽  
Tomoya Kawazoe ◽  
Salah M. El-Sayed ◽  
Kazuko Yorita ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Soluble Fraction ◽  
Recombinant Escherichia Coli ◽  
Acid Oxidase ◽  
Bioprocess Development ◽  
Amino Acid Oxidase

scholarly journals Porcine D-amino acid oxidase: Production of the biologically active enzyme in Escherichia coli

1989 ◽  
Vol 161 (2) ◽  
pp. 865-872 ◽  
Author(s):  
E. Ciccarelli ◽  
M. Massaer ◽  
J.-P. Guillaume ◽  
A. Herzog ◽  
R. Loriau ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Active Enzyme ◽  
Biologically Active ◽  
Acid Oxidase ◽  
Amino Acid Oxidase

scholarly journals EXPRESSION OF GLUTARYL7-AMINOCEPHALOSPORANIC ACID ACYLASE IN ESCHERICHIA COLI BL21(DE3) AND IMMOBILIZATION OF RECOMBINANT ENZYME ON NANOPOROUS MATERIALS

2018 ◽  
Vol 54 (4A) ◽  
pp. 123
Author(s):  
Vu Thi Hanh
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Expression Vector ◽  
Nanoporous Materials ◽  
Acid Oxidase ◽  
Nano Materials ◽  
Amino Acid Oxidase ◽  
E Coli ◽  
Escherichia Coli Bl21 ◽  
Gla Gene

The synthesis of 7-ACA from cephalosporin C (CPC) by a two-step bioconversion using D-amino acid oxidase (DAAO) and glutaryl 7-ACA acylase (GLA) has been effectively and largely applied in pharmaceutical industry. In this study, the gene gla coding for 720-amino acid GLA from plasmid pUC57::gla was analyzed and successfully inserted into vector pET22b(+) to form expression vector pET22b(+)::gla. The newly constructed expression vector pET22b(+)::gla was cloned and then transformed into Escherichia coli BL21(DE3) to generate recombinant strain E. coli BL21(DE3)[pET22b(+)::gla]. The suitable conditions for expression of gla gene were in LB medium at 30 oC and induced by 0.4 mM of Isopropyl β-D-1-thiogalactopyranoside (IPTG) for 3 hours. Under the chosen culturing parameters, expression of gla gene by E. coli BL21(DE3)/[pET22b(+)::gla] resulted in a recombinant GLA (rGLA) with molecular weight of 83 kDa and catalytic activity of 2.7 U/mg of total protein. Experimental research on immobilization of rGLA onto ten nanoporous materials were showed that, SBA-15 was the best one for immobilization of rGLA, reaching activity of immobilized enzyme of 22.2 U/g matrix. Furthermore, optimal conditions of procedure for immobilizing rGLA on nanomaterials (SBA-15) were determined as follows: temperature is 25 °C, pH7.0 and immobilization time –60 minutes. Therefore the results reported in this study revealed the successfully heterologous expression of GLA in recombinant E. coli and potential immobilization of enzyme on inorganic nano-materials.

scholarly journals Cloning, expression and purification of fructosyl amino acid oxidase (FAOX) in Escherichia Coli

2021 ◽  
Vol 11 (1) ◽  
pp. 21-29
Author(s):  
Ho Ta Giap ◽  
Phan Ngoc Han ◽  
Tran Le Duy Phuong ◽  
Phung Thi Thu Phung ◽  
Vu Van Van
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Exchange Chromatography ◽  
Cloning And Expression ◽  
Future Application ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Colony Pcr ◽  
E Coli ◽  
Fructosyl Amino Acid Oxidase

Introduction: The level of serum HbA1c is an indicator of the average blood sugar level in the last three months. HbA1c can be quantified using assays involving the enzyme fructosyl amino acid oxidase (FAOX). This study aims to produce GST-tagged FAOX-TE (GST/FAOX-TE), a thermal stable and specific variant of FAOX, for future application studies. Materials and methods: The E. coli strains DH5α and BL21 (DE3) were used as cloning and expression hosts, respectively. The FAOX-TE sequence was synthesized at IDT (US) and clonned into pGEX-4T3 vector, which was confirmed by Colony PCR. The expression was induced at 16°C, 0.5 mM IPTG in LB media containing 50 µg/ml ampicilin. The protein expression profile was analyzed by SDS-PAGE. The cell pellet was sonicated and purified by Glutathione Sepharose 4 Fast Flow (Cytiva, US). The catalytic activity of GST/FAOX-TE with fructosyl valine was determined using high performance anion exchange chromatography with pulsed amperometry detection (HPAEC-PAD). Results: The fusion protein was successfully expressed in Escherichia coli using the plasmid pGEX-4T3 and purified to high purity 93%. Recombinant GST/FAOX-TE was shown to be active on fructosyl valine. Conclusions: Active GST/FAOX-TE was successfully expressed in E. coli BL21 (DE3) and purified, which will be used for future development of biosensors for fructosyl valine quantification.

scholarly journals Trigonopsis variabilis d-Amino Acid Oxidase: Control of Protein Quality and Opportunities for Biocatalysis through Production in Escherichia coli

2006 ◽  
Vol 73 (1) ◽  
pp. 331-333 ◽  
Author(s):  
Iskandar Dib ◽  
Damir Stanzer ◽  
Bernd Nidetzky
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Protein Quality ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Original Activity ◽  
Toxic Activity ◽  
Enzyme Preparations ◽  
Trigonopsis Variabilis ◽  
Natural Enzyme

ABSTRACT Trigonopsis variabilis d-amino acid oxidase accounts for 35% of Escherichia coli protein when added d-methionine suppresses the toxic activity of the recombinant product. Permeabilized E. coli cells are reusable and stabilized enzyme preparations. The purified oxidase lacks the microheterogeneity of the natural enzyme. Oriented immobilization of a chimeric oxidase maintains 80% of the original activity in microparticle-bound enzymes.

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