$GA_3$에 의하여 유도된 $alpha$-amylase (대조구의 $alpha$-amylase)와 $GA_3$ 처리 12시간 경과 후 canavanine을 첨가하여 얻은 $alpha$-amylase (canavanine 처리구의 $alpha$-amylase)를 ammonium sulfate분획, CHA-affinity chromatography와 DEAE-cellulose chromatography를 사용하여 정제하였고 이들 두 효소의 성질을 비교 조사하였다. Canavanine 처리구의 $alpha$-amylase를 아마노산 분석한 결과 $alpha$-amylase 내에 1.5~1.7 mol의 canavanine이 투입되었음이 확인되었다. 대조구의 $alpha$-amylase와 canavanine 처리구의 $alpha$-amylase의 pI값은 각각 5.2와 4.9였다. 그러나 분자량은 50,000 dalton으로 동일하게 나타났으며, 최적온도, 최적 pH, 그리고 반응산물도 동일하였다. 그러나 amylose를 기질로 사용하였을 때의 최대반응속도는 대조구의 $alpha$-amylase의 경우 canavamne 처리구의 $alpha$-amylase의 경우보다 1.6배 빨랐다. 그러나 $K_m$값은 동일하였다. 또한 $Ca^{2+}$, $Mg^{2+}$ 및 $Mn^{2+}$과 같은 $alpha$-amylase의 activator를 첨가하였을 예는 대조구의 $alpha$-amylase의 경우 $10^{-3}M$보다 $10^{-2};M$처리시 그 활성이 높게 나타났으나 canavanine 처리구의 $alpha$-amylase의 경우 $10^{-3};M$ 보다 오히려 그 활성이 감소하였다. 이상의 결과를 종합해 보면 보리종자에 canavanine을 처리하였을 때 canavanine이 $alpha$-amylase내로 투입되어 효소구조를 변화시킴으로써 효소활성을 감소시키는 것으로 사료된다.
The effect of canavanine on the $GA_3$-induced $alpha$-amylase in barley halfseeds were investigated. $GA_3$-induced $alpha$-amylase (control $alpha$-amylase) and the $alpha$-amylase to which canavanine was added 12 h after $GA_3$ treatment (canavanine treated $alpha$-amylase) were purified through ammonium sulfate fractionation, CHA-affinity chromatography and DEAE-cellulose chromatography, and the properties of both kinds of enzyme were comparatively studied. The amino acid analysis of canavanine treated $alpha$-amylase revealed that 1.5-1.7 mol of canavanine was incorporated into $alpha$-amylase instead of arginine. The pI values of control $alpha$-amylase and canavanine treated $alpha$-amylase were 5.2 and 4.9 respectively. However, their molecular weights were determined as the same dalton of 50,000 and their optimum temperature and optimum pH as well as the reaction products were the same in both kinds. While $K_m$ values were the same in both kinds, the maximum enzyme reaction velocity using amylose as substrate were 1.6 times faster for control $alpha$-amylase than for canavanine treated $alpha$-amylase. When the enzyme activators such $Ca^{2+}$, $Mg^{2+}$, and $Mn^{2+}$ were treated upon the purified enzymes, the activator concentration of $10^{-3};M$ had less effect on the activation of control $alpha$-amylase then the concentration of $10^{-2};M$. However, the situation was reversed in the case of canavanine treated $alpha$-amylase. In summary, when canavanine was treated upon barley seeds, its incorporation into $alpha$-amylase is considered to decrease the $alpha$-amylase activity through. changing the structure of the enzyme.
