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Volumn 3, Issue 6, 2003, Pages 829-832

Enzyme-polymer-single walled carbon nanotube composites as biocatalytic films

Author keywords

[No Author keywords available]

Indexed keywords

CARBON; CHYMOTRYPSIN A; ENZYME; GRAPHITE; POLY(METHYL METHACRYLATE); POLYMER; TOLUENE;

EID: 0141521836     PISSN: 15306984     EISSN: None     Source Type: Journal    
DOI: 10.1021/nl034131k     Document Type: Article
Times cited : (162)

References (23)
  • 20
    • 0141513346 scopus 로고    scopus 로고
    • note
    • One gram of polymer was dissolved in 3.0 g (∼3.5 mL) toluene by ultrasonicating for 5 min and allowed to cool to room temperature. A predetermined weight of SWNT (Carbon Nanotechnologies, Inc., Houston, TX) was added to 1.0 g toluene and the dispersion was sonicated until a uniform suspension of the SWNT was obtained and allowed to cool to room temperature. One milligram of α-chymotrypsin (Sigma Chemical Co., St. Louis, MO) was added, and the resulting suspension was sonicated for 1 min to ensure a uniform dispersion of both the enzyme and the SWNT. After cooling to room temperature, the polymer - toluene solution and the enzyme - SWNT - toluene suspensions were mixed together and drop cast into a 5-cm Petri dish to form a composite film. Similar approaches were used for the graphite (avg. particle size ≤ 100 μm from Fluka Chemie AG, Switzerland) and additive-free enzyme-pMMA films or for other polymer films.
  • 21
    • 0141624649 scopus 로고    scopus 로고
    • note
    • CT was chosen as the model enzyme for all films studied and N-succinyl Ala-Ala-Pro-Phe-p-nitroanilide was used as a chromogenic substrate. The tetrapeptide substrate is cleaved by CT to yield the p-nitroaniline chromophore, which was detected spectrophotometrically at 410 nm (Perkin-Elmer Lambda 6.0 UV-vis spectrophotometer (Norwalk, CT)). To study the leaching of CT from the films, repeated washing was performed with 4 mL of 20 mM Tris buffer, pH 7.4. Each wash carried out for 12 min in an orbital shaker at 135 rpm and 25 °C. After successive washes, the supernatants were collected and filtered, and 25 μL of the tetrapeptide substrate solution was added at a final concentration of 60 μM. The films were washed until no further increase in leachate enzyme activity was obtained or a stable release of enzyme activity was measured. To measure enzyme activity specifically in the film, the leached enzyme solution was replaced with a fresh substrate solution in aqueous buffer and the observed enzyme activity determined. The "final film activity" or "film activity" was calculated by subtracting the final wash activity from the observed film activity.
  • 23
    • 0141513330 scopus 로고    scopus 로고
    • note
    • 20 and then washed five times with DI water by pouring water onto the films and shaken at 100 rpm for 10 min. This was followed by cutting a center section from the film for DSC analysis. The remaining part of each film was placed in water for 48 h and then subjected to an identical sequence of washing and DSC analysis as described above.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.