Modification of Papain with Soluble Starch

Periodate-oxidized soluble starch was reacted with papain at pH 4.0, pH 7.0, and pH 9.7, and an oxidized soluble starch-papain conjugate was produced. When compared with native papain, the specific activity decreased to 60%, in both the modified papain reacted with 0.4% $NaBH_4$ and in the modified papain not reacted with $NaBH_4$. The specific activity decreased to 70% in the modified papains reacted with 1.5% $NaBH_4$ and 4.0% $NaBH_4$, respectively. The reduction by $NaBH_4$ did not have an effect in the thermal stability of either the modified or nonmodified papain. An activity of 54.7% remained in the papain modified at pH 4.0, which was incubated at $80^{\circ}C$ for 40 min. The papains modified at pH 7.0 and pH 9.7 and incubated for 40 min at two different temperatures, respectively, were stable to $60^{\circ}C$, and at $80^{\circ}C$ their activities at 56.3% and 44.1 %, respectively. The modified papain's thermal stability pattern was similar to that of native papain, with no increase in its statbility. In the range of pH $2.0{\sim}13.0$, the stability of the papain modified at pH 4.0 decreased greatly between pH $3.0{\sim}5.0$, but it was similar to the native papain at other pH values. The stability of the papain modified at pH 7.0 showed a similar pattern to the native papain at pH $2.0{\sim}6.0$, while its stability increased when moving into the alkali pH range. The papain modified at pH 9.7 also had increased stability, when moving into the alkali range. The results of Hammerstein milk casein, which was reacted with the papains modified at pH 4.0, pH 7.0, and pH 9.7, respectively, and analyzed by FPLC, showed different peaks according to the different modification pHs, and the greatest peak differences were observed with the modification at pH 9.7.

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