Title: Optimization of Baccatin III Production by Cross-Linked Enzyme Aggregate of Taxoid 10β-O-Acetyltransferase.

Authors: You, Lin-Feng; Wei, Tao; Zheng, Qian-Wang; Jiang, Bing-Hua; Lin, Jun-Fang; Guo, Li-Qiong

Published In Mol Biotechnol, (2019 Jul)

Abstract: Taxoid 10β-O-acetyltransferase (DBAT) is the key enzyme to produce baccatin III, a key precursor in paclitaxel synthesis, by acetyl group transfer from acetyl-CoA to the C10 hydroxyl of 10-deacetylbaccatin III. In this study, the recombinant DBAT (rDBAT) was immobilized by cross-linked enzyme aggregates (CLEAs). To further optimize the enzyme recovery, single-factor experiment and response surface methodology were applied. 60% ammonium sulfate as precipitant, 0.05% glutaraldehyde as fixing agent, pH 7.0, 2 h as cross-linking time, 30 °C as cross-linking temperature were confirmed to be the optimum conditions to prepare the CLEAs-rDBAT in single-factor experiment. In addition, 62% for ammonium sulfate saturation, 0.15% for glutaraldehyde, and pH 6.75 were confirmed to be the optimum conditions with averagely 73.9% activity recovery in 3 replications, which was consistent with the prediction of response surface methodology. After cross-linking, the optimum temperature of CLEAs-rDBAT rose up to 70 °C and CLEAs-rDBAT could be recycled for three times.

PubMed ID: 31119478

MeSH Terms: Acetyltransferases/biosynthesis*; Acetyltransferases/genetics; Alkaloids/biosynthesis*; Alkaloids/genetics; Cross-Linking Reagents; Enzymes, Immobilized/biosynthesis*; Paclitaxel/biosynthesis; Recombinant Proteins/biosynthesis*; Taxoids