FOOD SCIENCE ›› 2017, Vol. 38 ›› Issue (2): 7-14.doi: 10.7506/spkx1002-6630-201702002

• Bioengineering • Previous Articles     Next Articles

Gene Cloning, Expression and Characterization of Protein Disulfide Isomerase from Wheat (Triticum aestivum L.)

LIU Guang, HU Songqing, ZHANG Tingting, WANG Jingjing, LI Lin, HOU Yi   

  1. 1. School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China; 2. Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, Guangzhou 510640, China
  • Online:2017-01-25 Published:2017-01-16

Abstract: Purpose: The gene encoding wheat protein disulfide isomerase (wPDI) was cloned and expressed in Escherichiacoli, and its enzymatic properties were investigated. Methods: The wpdi gene was obtained by reverse transcriptionpolymerase chain reaction amplification using the total RNA from wheat seeds as template. The recombinant plasmid pET-30b-wpdi was constructed and transformed into E. coli BL21 (DE3). After metal chelating chromatography, the enzymaticproperties of the purified wPDI were determined. Results: A 1 548 bp gene fragment was amplified and sequenced as wpdigene that had 99% identity with that of the wheat cultivar Wyuna. The optimized conditions for wPDI expression weredetermined as follows: induction at 22 ℃ using isopropyl β-D-1-thiogalactopyranoside at a concentration of 0.5 mmol/L for6 h. The recombinant wPDI consisted of four thioredoxin-like domains and had a molecular weight of 66.2 kD. The enzymeexhibited enzymatic activities (including reductase activity and isomerase activity of disulfide bonds) and chaperone activity.Conclusions: The expression of wPDI and its enzymatic properties can provide the foundation for its application in the flourprocessing industry.

Key words: wheat protein disulfide isomerase, cloning, expression, enzymatic activities

CLC Number: