碩士 / 國立清華大學 / 生命科學系 / 87 / Recombinant D-hydantoinase (AH) from Agrobaterium radiobacter DH102 was used to produce N-carbamoyl-D-p-hydroxyphenylglycin (NC-HPG) from D,L-hydroxyphenylhydantoin (D,L-HPH).
Determination of the relative molecular mass of the protein by gel-filtration chromatography gave an
apparent value of 101.57 kD. The subunit Mr was 56 kD, as estimated by analytical SDS/PAGE. The
enzyme was purified by a procedure including streptomycin sulfate treatment, ammonium sulfate
fractionation, six chromatographies, and was partially purified to about 90% purity on SDS-PAGE.
Some biochemical properties were characterized. The optimal growth temperature of the bacterium
was about 28℃. The optimal pH and temperature of recombinant D-hydantoinase were determined to
be about 8.0 and 55℃, respectively. And the half-life of the enzyme was about 5hr at 37℃. The purified
recombinant D-hydantoinase was not stable even at 4℃, however. Our results of the experiments showed
the low activity and stability of the enzyme, suggesting further useful strategy should be attempted.
| Identifer | oai:union.ndltd.org:TW/087NTHU0105005 |
| Date | January 1998 |
| Creators | Wen-Huey Cheng, 鄭文惠 |
| Contributors | Tzong-Hsiung Hseu, 許宗雄 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 63 |
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