2019 ; Vol.29-12: 1882~1893
|Author||Qi Li, Tao Wu, Linguo Zhao, Jianjun Pei, Zhenzhong Wang, Wei Xiao|
|Place of duty||College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, Nanjing 210037, China,Jiangsu Key Lab for the Chemistry & Utilization of Agricultural and Forest Biomass, 159 Long Pan Road, Nanjing 210037, China|
|Title||Highly Efficient Biotransformation of Astragaloside IV to Cycloastragenol by Sugar-Stimulated モ-Glucosidase and モ-Xylosidase from Dictyoglomus thermophilum|
J. Microbiol. Biotechnol.2019 ;
|Abstract||モ-Glucosidases and モ-xylosidases are two categories of enzymes that could cleave out nonreducing,
terminal モ-D-glucosyl and モ-D-xylosyl residues with release of D-glucose and Dxylose,
respectively. In this paper, two functional モ-glucosidase Dth3 and モ-xylosidase Xln-DT
from Dictyoglomus thermophilum were heterologously expressed in E.coli BL21 (DE3). Dth3 and
Xln-DT were relatively stable at 75oC and were tolerant or even stimulated by glucose and
xylose. Dth3 was highly tolerant to glucose with a Ki value of approximately 3 M. Meanwhile,
it was not affected by xylose in high concentration. The activity of Xln-DT was stimulated 2.13-
fold by 1 M glucose and 1.29-fold by 0.3 M xylose, respectively. Furthermore, the モ-
glucosidase Dth3 and モ-xylosidase Xln-DT showed excellent selectivity to cleave the outer C-6
and C-3 sugar moieties of ASI, which established an effective and green method to produce
the more pharmacologically active CAG, an exclusive telomerase activator. We measured
temperature, pH and dosage of enzyme using a single-factor experiment in ASI
biotransformation. After optimization, the optimal reaction conditions were as follows: 75oC,
pH 5.5, 1 U of Dth3 and 0.2 U of Xln-DT, respectively. Under the optimized conditions, 1 g/l
ASI was transformed into 0.63 g/l CAG with a corresponding molar conversion of 94.5%
within 3 h. This is the first report to use the purified thermostable and sugar-tolerant enzymes
from Dictyoglomus thermophilum to hydrolyze ASI synergistically, which provides a specific,
environment-friendly and cost-effective way to produce CAG.|
|Key_word||Glycosidases, sugar-tolerant, biotransformation, astragaloside IV, cycloastragenol|
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