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Improvements of Modified Wheat Protein Disulfide Isomerases with Chaperone Activity Only on the Processing Quality of Flour

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Abstract

Wheat protein disulfide isomerase (wPDI) with oxidoreductase activity, isomerase activity, and chaperone activity catalyzes the formation of disulfide bonds in gluten in vitro; the addition of wild-type wPDI weakened yet the flour processing quality. To develop the potential flour improvers from wPDI, the modified wPDIs were prepared by the biological or chemical method, and their effects on the processing quality of flour in connection with bread making were investigated by farinograph, texture profile analysis, electrophoresis, size exclusion chromatography, and scanning electron microscopy. A truncated protein of wPDI, fragment AB with oxidoreductase, and isomerase activities only was firstly confirmed to exert deteriorative effects similar to wPDI on the dough and bread quality. Then other two modified wPDIs, mPDI and aPDI, were prepared by site-directed mutagenesis and alkylation, respectively. Both mPDI and aPDI considerably retained the chaperone activity of wPDI but completely lost oxidoreductase and isomerase activities. After adding the appropriate amount of mPDI or aPDI, the stability time of dough was significantly prolonged from 3.60 to 4.15 or 4.13 min, respectively. The enhanced gluten network matrix and decreased hardness and chewiness of bread further suggested that dough was strengthened by the treatment of mPDI or aPDI. Moreover, the formation of gluten network was facilitated by the modified wPDIs with chaperone activity only for the increase amount of gluten macropolymer and the decrease content of SDS-soluble gluten. Consequently, mPDI and aPDI are valid candidates of flour improvers in food industry.

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Abbreviations

PDI:

Protein disulfide isomerase

hPDI:

Human protein disulfide isomerase

hPDIp:

Human pancreas-specific protein disulfide isomerase

wPDI:

Wheat protein disulfide isomerase

A:

Domain a of wPDI

AB:

Domains a and b of wPDI

mPDI:

Site-directed mutant wPDI cysteine residues in both active sites replaced by serine residues

aPDI:

Alkylated wPDI cysteine residues in both active sites alkylated by iodoacetamide

DS:

Degree of softening

WA:

Water absorption

DDT:

Dough development time

DST:

Dough stability time

FU:

Farinograph units

GMP:

Gluten macropolymer

GRAS:

Generally recognized as safe

ER:

Endoplasmic reticulum

RT-PCR:

Reverse transcription-polymerase chain reaction

Cys:

Cysteine

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

DTT:

Dithiothreitol

PBS:

Phosphate

EDTA:

Ethylene diamine tetraacetic acid

HMW-GS:

High molecular weight glutenin subunits

LMW-GS:

Low molecular weight glutenin subunits

RNase:

Ribonuclease

rRNase:

Inactive ribonuclease with reduced disulfide bonds

sRNase:

Inactive ribonuclease with scrambled disulfide bonds

2′3′-cCMP:

Cytidine 2′,3′-cyclic monophosphate

3′-CMP:

Cytidine 3′-monophosphate

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SEM:

Scanning electron microscope

BSA:

Bovine serum albumin

TPA:

Texture profile analysis

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Acknowledgements

We appreciate financial support from the National Natural Science Foundation of China (grant numbers 31471691, 31171630, and 31130042), the Specialized Research Fund for the Doctoral Program of Higher Education of China (number 20130172110018), the Science and Technology Planning Project of Guangdong Province, China (grant number 2014A010107002), and the Science and Technology Planning Project of Foshan city, Guangdong Province, China (grant number 2015AG10011).

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Authors and Affiliations

  1. School of Food Sciences and Engineering, South China University of Technology, Guangzhou, Guangdong, 510641, China

    Guang Liu, JingJing Wang, Yi Hou, Lin Li & Song-Qing Hu

  2. State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou, Guangdong, 510640, China

    Yi Hou & Yan-Bo Huang

  3. Department of Food Science and Engineering, Jinan University, Guangzhou, Guangdong, 510632, China

    Cun-Zhi Li

  4. Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, South China University of Technology, Guangzhou, Guangdong, 510640, China

    Lin Li & Song-Qing Hu

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  1. Guang Liu
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Liu, G., Wang, J., Hou, Y. et al. Improvements of Modified Wheat Protein Disulfide Isomerases with Chaperone Activity Only on the Processing Quality of Flour. Food Bioprocess Technol 10, 568–581 (2017). https://doi.org/10.1007/s11947-016-1840-9

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