Comparison between Polyvinyl Pyrrolidone/Na2SO4 Aqueous Two-Phase Systems and Chromatographic Methods for Purification of Recombinant Phenylalanine Dehydrogenase
Phenylalanine dehydrogenase (PheDH; EC 1.4.1.20) is an important enzyme of amino acid dehydrogenases family that increasingly used as a valuable biocatalyst in neonatal screening kits and synthesis of L-phenylalanine. The goal of this literature was to find a suitable purification method for recombi...
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| Main Author: | |
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| Format: | Article |
| Language: | English |
| Published: |
University of Tehran
2009-12-01
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| Series: | Journal of Sciences, Islamic Republic of Iran |
| Subjects: | |
| Online Access: | https://jsciences.ut.ac.ir/article_20127_c4f3ede9d437423f7e3644fe92a55bb1.pdf |
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| Summary: | Phenylalanine dehydrogenase (PheDH; EC 1.4.1.20) is an important enzyme of amino acid dehydrogenases family that increasingly used as a valuable biocatalyst in neonatal screening kits and synthesis of L-phenylalanine. The goal of this literature was to find a suitable purification method for recombinant Bacillus badius PheDH by practical comparison between chromatographic and polyvinyl pyrrolidone (PVP)/Na2SO4 aqueous two-phase systems (ATPS) techniques. The partitioning behavior of target enzyme in PVP/Na2SO4 ATPS was examined and compared with the obtained results from a chromatographic protocol. Direct comparison of chromatography and ATPS procedures clearly revealed that the ATPS consisting of 8.0% (w/w) PVP, 17.0% (w/w) Na2SO4 with pH of 8.0, VR=0.25 and temperature of 25 °C was the most desirable process for PheDH purification. A specific activity of 1231.42 U/mg, a purification factor of 36.61, a yield of 95.5% and a recovery of 138.9% were achieved. Altogether, we presented a two-phase methodology as a scalable and economically alternative for the production of PheDH enzyme. |
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| ISSN: | 1016-1104 2345-6914 |