• BMB Reports
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• 제39권1호
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• pp.76-83
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• 2006

Pyridoxine-5-P oxidase catalyses the terminal step in the biosynthesis of pyridoxal-S-P, the biologically active form of vitamin $B_6$ Which acts as an essential cofactor. Here, a human brain pyridoxine-5-P oxidase gene was fused with a gene fragment encoding the HIV-1 Tat protein transduction domain (RKKRRQRRR) in a bacterial expression vector to produce a genetic in-frame Tat-pyridoxine-5-P oxidase fusion protein. Expressed and purified Tat-pyridoxine-5-P oxidase fusion protein transduced efficiently into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced Tat-pyridoxine-5-P oxidase protein showed catalytic activity and was stable for 48 h. Moreover, the formation of pyridoxal-5-P was increased by adding exogenous Tat-pyridoxine-5-P oxidase to media pre-treated with the vitamin $B_6$ precursor pyridoxine. In addition, the intracellular concentration of pyridoxal-S-P was markedly increased when Tat-pyridoxal kinase was transduced together with Tat-pyridoxine-5-P oxidase into cells. These results suggest that the transduction of Tat-pyridoxine-5-P oxidase fusion protein presents a means of regulating the level of pyridoxal-5-P and of replenishing this enzyme in various neurological disorders related to vitamin $B_6$.

• 한국식품과학회지
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• 제9권3호
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• pp.183-189
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• 1977

고정화(固定化) 균체(菌體)를 이용(利用)한 pyridoxal 5'-phosphate(pyridoxal-p)의 연속생산(連續生産)에 관(關)해실험(實驗)하였다. pyridoxine 5'-phosphate (pyridoxine-p) oxidase활성(活性)을 갖는 Pseudomonas polycolor 균체(菌體)와 Catalase 활성(活性)을 갖는 Kloeckera sp. No. 2201균체(菌體)를 효소원(酵素源)으로 사용(使用)하였다. 균체(菌體)는 Polyacrylamide gel에 불용화(不溶化) 시켰으며 동시고정균체(同時固定菌體)의 활성(活性)이 Pseudomonas Polycoler단독고정균체(單獨固定菌體)의 활성(活性)보다 강(强)하였다. 이 결과(結果)는 동시고정균체(同時固定菌體)의 Pyridoxine-p oxidase-cat-alase system은 Pyridoxine-p 산화(酸化)의 부산물(副産物)인 $H_2O_2$를 분해(分解)하므로서 보호효과를 얻을 수 있음을 의미(意味)한다. 동시고정균체(同時固定菌體)와 생균체(生菌體)의 효소활성(酵素活性)에 미치는 최적(最適)pH는 9.0이었고 동시고정균체(同時固定菌體)의 최적온도(最適溫度)는 $45^{\circ}C$로 생균체(生菌體)보다 $5^{\circ}C$높았다. 고정균체(固定菌體)의 Pyridoxine-p oxidase는 $Hg^{2+}$와 몇몇 SH-화합물(化合物)에 의(依)하여 활성화(活性化)되었다.