Expression of Codon-Optimized Plant Glycosyltransferase UGT72B14 in Escherichia coli Enhances Salidroside Production.
Salidroside, a plant secondary metabolite in Rhodiola, has been demonstrated to have several adaptogenic properties as a medicinal herb. Due to the limitation of plant source, microbial production of salidroside by expression of plant uridine diphosphate glycosyltransferase (UGT) is promising. However, glycoside production usually remains hampered by poor expression of plant UGTs in microorganisms. Herein, we achieved salidroside production by expression of Rhodiola UGT72B14 in Escherichia coli (E. coli) and codon optimization was accordingly applied. UGT72B14 expression was optimized by changing 278 nucleotides and decreasing the G+C content to 51.05% without altering the amino acid sequence. The effect of codon optimization on UGT72B14 catalysis for salidroside production was assessed both in vitro and in vivo. In vitro, salidroside production by codon-optimized UGT72B14 is enhanced because of a significantly improved protein yield (increased by 4.8-fold) and an equivalently high activity as demonstrated by similar kinetic parameters (K M and V max), compared to that by wild-type protein. In vivo, both batch and fed-batch cultivation using the codon-optimized gene resulted in a significant increase in salidroside production, which was up to 6.7 mg/L increasing 3.2-fold over the wild-type UGT72B14.
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Feiyan
F
Xue
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Huili
H
Guo
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Yingying
Y
Hu
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Ran
R
Liu
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Lina
L
Huang
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Heshu
H
Lv
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Chunmei
C
Liu
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Mingfeng
M
Yang
Key Laboratory of Urban Agriculture (North), Ministry of Agriculture, Beijing 102206, China; College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China; Beijing Collaborative Innovation Center for Eco-Environmental Improvement with Forestry and Fruit Trees, Beijing 102206, China.
Lanqing
0000-0002-5578-6958
L
Ma
D000595:Amino Acid Sequence / Q000235:genetics; D002384:Catalysis; D003062:Codon / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D005960:Glucosides / Q000096:biosynthesis* / Q000235:genetics; D016695:Glycosyltransferases / Q000096:biosynthesis* / Q000235:genetics; D010636:Phenols; D029771:Rhodiola / Q000235:genetics*
D003062:Codon; D005960:Glucosides; D010636:Phenols; D016695:Glycosyltransferases; C009172:rhodioloside