Proleather from Bacillus sp. was chemically modified with decanoyl chloride for enhanced activity for the preparation of poly(lactic acid) in organic solvents. The modified enzyme was highly soluble (up to 44 mg-protein/ml) and active in various organic solvents including chloroform, tetrahydrofuran (THF), pyridine and acetone. The organic-soluble proleather efficiently catalyzed the polymerization of ethyl lactate. The reaction rate was 4-22 times that of native proleather, depending upon the solvent applied. The solubilized enzyme showed a highest activity at 50°C, the same optimum temperature for both the native proleather and an immobilized enzyme, Novozyme-435. Denaturation of the enzymes' protein structures appeared to be the critical factor regulating the optimum activity temperature. Differential scanning calorimetry (DSC) analyses of the enzymes showed endothermic peaks around 55°C, indicating the proteins' structures altered in that temperature range. Interestingly, the activity of the solubilized enzyme showed a more complicated water dependence as compared to native proleather.
Bibliographical noteFunding Information:
This work was supported by grants from US Department of Agriculture (#2001-35504-10757), ACS Petroleum Research Fund (#36726-G4) and National Science Foundation (CTS-0124769). The authors thank Dr. Lu-Kwang Ju for valuable discussions and Mr. Hongfei Jia and Mr. Russel C. Vogelsong for assisting in the experimental work.
- Nonaqueous biocatalysis
- Poly(lactic acid)