Studies on the Formation of Pyridoxal Phosphate by Immobilized Cells

고정화 균체에 의한 Pyridoxl Phosphate의 생산에 관한 연구

Studies were made of the continuous production of Pyridoxal 5'-phosphate (pyridoxal-p) on simultaneously immobilized cell column. Whole-cell of Pseudomonas polycolor having high activity of pyridoxine 5'-phosphate (pyridoxine-p) oxidase and Kloeckera sp. No. 2201 having high activity of catalase were used as the enzyme materials. The enzyme sources were entrapped in a polyacrylamide gel. Enzymatic properties of the simultaneously immobilized cells were investigated, comparing with those of the mixed whole-cells of the microorganisms. The simultaneously immobilized cells had higher enzyme activity than singly immobilized cells of Pseudomonas polycolor. From this result, the simultaneously immobilized pyridoxine-p oxidase-catalase system could be available to exert a protective effect upon the pyridoxine-p oxidase by destroying $H_2O_2$ which is a by-product of pyridoxine-p oxidation. The optimum pH was 9.0 for the immobilized cells and the whole-cells. The optimum temperature was $45^{\circ}C$ for the immobilized cells and $40^{\circ}C$ for the whole-cells. The pyridoxine-p oxidase of the immobilized cells were activated by $Hg^{++}$ and some SH-compounds.

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.