Chaperone-Mediated Protein Folding Enhanced D-Psicose 3-Epimerase Expression In Engineered Bacillus Subtilis
PROCESS BIOCHEMISTRY(2021)
摘要
Functional mature proteins usually undergo folding, and in vivo folding is often affected by factors such as translation ratio, intracellular environment, and chaperones. D-psicose 3-epimerase (DPEase) derived from Dorea sp. has excellent properties in catalytic production of D-allulose; a rare sugar with unique biological functions. Here, the Dorea sp. DPEase coding gene was fusion-expressed with four native chaperones (DnaK, DnaJ, GrpE and PrsA) in Bacillus subtilis, and resulted in an 4.31- and 6.12-fold increase to 10.81 U/g and 13.34 U/g in recombinant cells of BS-DPE-DnaK and BS-DPE-PrsA, respectively. The recombinant proteins showed increased thermostability without significant change in the optimum pH and Co2+ affinity. Using recombinant cells as catalysts, a 1.12-2.31-fold increase was detected in DPEase activity after treatment with 40 % ethanol. We showed that fusion expression of molecular chaperones to enhance protein-assisted folding could facilitate expression of DPEase. We hope that our results provide a facilitated route for large-scale production of D-allulose in B. subtilis.
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关键词
D-psicose 3-epimerase, Bacillus subtilis, Chaperone, Whole-cell catalysis, Protein folding
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