Novel technique for high throughput measurement of active monooxygenase concentration
| Autor: | Anna Joëlle Ruff, Thomas Horn, Tim Welters, Jochen Büchs, Ulrich Schwaneberg |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Accuracy and precision Optical fiber Absorption spectroscopy Chemistry Analytical chemistry Bioengineering 010501 environmental sciences 01 natural sciences Applied Microbiology and Biotechnology law.invention Absorbance 03 medical and health sciences Wavelength 030104 developmental biology law Measuring principle Spectroscopy Throughput (business) 0105 earth and related environmental sciences Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 114:929-933 |
| ISSN: | 0006-3592 |
| DOI: | 10.1002/bit.26215 |
| Popis: | Absorbance measurements via transmitting light spectroscopy in microtiter plates are established for high throughput screening of biological systems. These measurements allow for the determination of important process parameters within a short time. However, absorbance determination via transmitted light measurements is not always feasible. As for carbon monoxide difference absorbance spectroscopy, used for concentration measurements of active P450 monooxygenases (P450s), security standards, and consistent gassing have to be addressed. In this study, a non-invasive online measuring principle for absorbance via scattered light is proposed. Based on optical fiber measurements, a decrease in scattered light signals at 450 nm wavelength of reflecting polymer particles is observed, and P450 concentrations are calculated. In this way, high throughput determination of P450 concentrations in a secure, gas-tight environment is realized. The designed method was successfully applied to concentration measurements and carbon monoxide (CO) saturation kinetics ranging from 0.3 to 5.0 μM P450 BM3 achieving a measurement accuracy of ±0.05 μM P450. Biotechnol. Bioeng. 2017;114: 929-933. © 2016 Wiley Periodicals, Inc. |
| Databáze: | OpenAIRE |
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