Characterization of lysine-tagged Bacillus stearothermophilus leucine aminopeptidase II immobilized onto carboxylated gold nanoparticles

Wu, Chia Ching; Chen, Yipu; Yang, Jia-Ci; Lo, Huei-Fen; Lin, Long-Liu

doi:10.1016/j.molcatb.2007.12.024

Cited by 17 publications

(10 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To produce BsAPII, BsAPII-Lys 3 , and BsAPII-Lys 9 , Escherichia coli M15 cells harboring pQE-LAPII [18], pQE-LAPII-Lys 3 [22], or pQE-LAPII-Lys 9 [22] were grown at 37°C in 200-ml Luria-Bertani medium supplemented with 100 g/ml ampicillin and 25 g/ml kanamycin to an absorbance at 600 nm of 1.0. Isopropyl--D-thiogalactopyranoside (IPTG) was added to a Wnal concentration of 0.5 mM and the incubation was continued at 28°C for 12 h. Cells were then harvested by centrifugation and resuspended in 10 ml of 50 mM Tris-HCl buVer (pH 8.0).…”

Section: Preparation Of Cell-free Extractsmentioning

confidence: 99%

See 1 more Smart Citation

Preparation of carboxylated magnetic particles for the efficient immobilization of C-terminally lysine-tagged Bacillus stearothermophilus aminopeptidase II

Huang

Wei

Yang

et al. 2010

J Ind Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

This article reports the synthesis and use of surface-modified iron oxide particles for the simultaneous purification and immobilization of Bacillus stearothermophilus aminopeptidase II (BsAPII) tagged C-terminally with either tri- or nona-lysines (BsAPII-Lys(3/9)). The carboxylated magnetic particles were prepared by the simple co-precipitation of Fe(3+)/Fe(2+) in aqueous medium and then subsequently modified with adipic acid. Transmission electron microscopy (TEM) micrographs showed that the carboxylated magnetic particles remained discrete and had no significant change in size after binding BsAPIIs. Wild-type enzyme and BsAPII-Lys(3) could be purified to near homogeneity by the carboxylated magnetic particles, but it was not easy to elute the adsorbed BsAPII-Lys(9) from the matrix. Free BsAPII, BsAPII-Lys(3), and BsAPII-Lys(9) were active in the temperature range 50-70 degrees C and all had an optimum of 50 degrees C, whereas the optimum temperature and thermal stability of BsAPII-Lys(3) and BsAPII-Lys(9) were improved as a result of immobilization. The immobilized BsAPII-Lys(9) could be recycled ten times without a significant loss of the enzyme activity and had a better stability during storage than BsAPII. Owing to its high efficiency and cost-effectiveness, this magnetic adsorbent may be used as a novel purification-immobilization system for the positively charged enzymes.

show abstract

Section: Preparation Of Cell-free Extractsmentioning

confidence: 99%

“…In our previous studies, three expression plasmids (pQE-LAPII, pQE-LAPII-Lys 3 , and pQE-LAPII-Lys 9 ) were constructed for the heterologous production of BsAPII, BsAPII-Lys 3 , and BsAPII-Lys 9 in E. coli [18,22]. The mRNAs of these constructs encode a His 6 tag at their N-termini.…”

Section: Adsorption Of the Engineered Enzymesmentioning

confidence: 99%

Preparation of carboxylated magnetic particles for the efficient immobilization of C-terminally lysine-tagged Bacillus stearothermophilus aminopeptidase II

Huang

Wei

Yang

et al. 2010

J Ind Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Ag and Au NPs have good electronic properties, and larger and stable surface area for enzyme immobilization [24]. They easily absorbed protein molecules on their surface without much activity loss and conformational changes [25,26].…”

Section: Introductionmentioning

confidence: 99%

Immobilization of yeast alcohol dehydrogenase on polyaniline coated silver nanoparticles formed by green synthesis

Alam

Laskar

Zubair

et al. 2015

Journal of Molecular Catalysis B: Enzymatic

View full text Add to dashboard Cite

“…In the visualizing method, the substrate also contains two main components. The first one is an enzyme‐specific artificial leucine and the second is the diazonium ion including naphthol and naphthylamine and their derivatives (Acartürk et al ., ; Prescott and Wilkes, ; Wu et al ., ). Either naphthol or naphthylamine is liberated enzymatically, and further, it couples with tetrazotized chromogens such as di‐ o ‐anisidine (diazo bulue B), N ‐(1‐naphthyl) ethylenediamine dihydrochloride or a modified Bratton–Marshall reaction and finally produces a colored azo complex.…”

Section: Introductionmentioning

confidence: 97%

Application of capillary electrophoresis coupling with electrochemiluminescence detection to estimate activity of leucine aminopeptidas

Ming

Wei

Zhou

et al. 2013

Biomedical Chromatography

View full text Add to dashboard Cite

A new method to estimate the leucine aminopeptidase (LAP, EC 3.4.11.1) activity using capillary electrophoresis coupled with electrochemiluminescence (ECL) is described. The liberated proline produced by LAP catalyzing the hydrolysis reaction of leucin-proline was used as an ECL coreagent to enhance Ru(bpy)3 (2+) ECL signals efficiently. The detection limit for proline was 2.88 × 10(-6) m (signal-to-noise ratio 3), which was equal to 9.60 × 10(-8) units of LAP being used to catalyze leucin-proline for 1 min. The Michaelis constant Km (2.07 × 10(-2) mol/L) and the maximum reaction velocity Vmax (1.06 × 10(-5) mol/L/min) of LAP for leucin-proline are reported. The reaction conditions including the concentration of metal ions, incubation temperature and pH were optimized. This method was successfully applied to detect LAP activity in plasma and the results were in good agreement with that obtained by the clinical method.

show abstract

Characterization of lysine-tagged Bacillus stearothermophilus leucine aminopeptidase II immobilized onto carboxylated gold nanoparticles

Cited by 17 publications

References 45 publications

Preparation of carboxylated magnetic particles for the efficient immobilization of C-terminally lysine-tagged Bacillus stearothermophilus aminopeptidase II

Preparation of carboxylated magnetic particles for the efficient immobilization of C-terminally lysine-tagged Bacillus stearothermophilus aminopeptidase II

Immobilization of yeast alcohol dehydrogenase on polyaniline coated silver nanoparticles formed by green synthesis

Application of capillary electrophoresis coupling with electrochemiluminescence detection to estimate activity of leucine aminopeptidas

Contact Info

Product

Resources

About