Heterologous expression and protein engineering of wheat gluten proteins

Tamás, László; Shewry, Peter R.

doi:10.1016/j.jcs.2006.02.001

Cited by 21 publications

(16 citation statements)

References 96 publications

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“…The direct addition of isolated native or heterologously expressed wheat storage proteins15 to a carefully chosen wheat flour (called base flour) allows the role of monomeric prolamins on rheological properties of the dough to be studied 16–19. However, if the added proteins are able to form polymers, simple addition does not show their real impact on the functional characteristics of the dough, because they do not become part of the polymeric network of the gluten 20–22.…”

Section: Introductionmentioning

confidence: 99%

Effects of incorporated amaranth albumins on the functional properties of wheat dough

et al. 2009

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BACKGROUND: Recent developments in micro-scale testing methodology and in methods modelling the effects of native forms of constituents by in vitro methods have provided a new approach to study the impact of added foreign proteins on dough end-use quality. Amaranth (Amaranthus) is a member of the pseudo-cereal family, whose storage proteins have superior nutritional quality due to their essential amino acid composition. The aim of this project was to study the effects of the incorporated amaranth albumin proteins on the rheological properties of the wheat dough.

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Section: Introductionmentioning

confidence: 99%

Effects of incorporated amaranth albumins on the functional properties of wheat dough

et al. 2009

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“…Three types of expression systems are widely used in heterologous protein production, among which pET series of vectors and E. coli were considered to be sui for the heterologous expression of wheat storage genes [15]. Despite of many improvements in expression vectors including combining strong promoters with suitable length, stringent regulators, transcription start sites, alternative translation start codons and multiple cloning site and transcription terminators, it is still difficult to express LMW-GS and gliadin genes in these vectors.…”

Section: Discussionmentioning

confidence: 99%

“…In the past several decades, three main types of expression systems have been widely used for heterologous protein production, viz. Escherichia coli, yeasts and cultured insect cells [15], among which the most popular for wheat storage protein gene expression is the E. coli because of its flexibility, ease of use and low cost [15][16][17].…”

Section: Background and Introductionmentioning

confidence: 99%

High-level expression of LMW-GS and α-gliadin genes promoted by the expressed tag sequence of 5′ end in Escherichia coli

Wang

Gao

et al. 2015

Protein Expression and Purification

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. AbstractWheat storage protein genes, especially low molecular weight glutenin subunit (LMW-GS) and gliadin genes are difficult to be expressed in E. coli, mainly due to the presence of highly repetitive sequences. In order to establish a high efficiency expression system for these genes, five different expression plasmids combining with 2 9 genes, viz. 6 LMW-GS and 3 α-gliadin genes isolated from common wheat and related species, were studied for heterologous expression in E. coli. In this study, when an expressed tag sequence encoding signal peptide, His-S or GST-tag was fused to the 5'end of LMW-GS or gliadin gene as the leading sequence, all recombination genes could be stably expressed at a high level. On the contrast, as expected, the inserted genes encoding mature protein failed without an expressed tag sequence. This result indicated that using expressed tag sequences as leading sequences could promote LMW-GS and gliadin genes to be well expressed in E. coli. Further transcriptional analysis by quantitative real-time PCR (qRT-PCR) showed transcription levels of recombination genes (e.g. GST-Glutenin, His-S-Glutenin and SP*-His-Glutenin) were 4-fold to 33-fold higher than those of the LMW-GS genes, which suggested these expressed tag sequences might play an important role in stimulating transcription. The possible molecular mechanism under this phenomenon was discussed.

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“…The three important things which are important for expression of gluten protein are expression system (E.coli, yeasts Generally, E.coli expression systems were suggested as suitable systems for many applications, because of following advantages (Tamas and Shewry 2006).Inducible promoters (with expression being induced by either specific chemicals or thermal effects) allow cultures to grow to high density providing higher yield and minimising effects of toxic proteins. The T7-based promoter induced by IPTG (isopropyl-b-D-thiogalactopyranoside) is particularly popular for high-level expression.…”

Section: (Ii) Gluten Proteinsmentioning

confidence: 99%

Protein engineering and its applications in food industry

Kapoor

Rafiq

Sharma

2015

Critical Reviews in Food Science and Nutrition

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Protein engineering is a young discipline that has been branched out from the field of genetic engineering. Protein engineering is based on the available knowledge about the proteins structure/function(s), tools/instruments, software, bioinformatics database, available cloned gene, knowledge about available protein, vectors, recombinant strains and other materials that could lead to change in the protein backbone. Protein produced properly from genetic engineering process means a protein that is able to fold correctly and to do particular function(s) efficiently even after being subjected to engineering practices. Protein is modified through its gene or chemically. However, modification of protein through gene is easier. There is no specific limitation of Protein Engineering tools; any technique that can lead to change the protein constituent of amino acid and result in the modification of protein structure/function is in the frame of Protein Engineering. Meanwhile, there are some common tools used to reach a specific target. More active industrial and pharmaceutical based proteins have been invented by the field of Protein Engineering to introduce new function as well as to change its interaction with surrounding environment. A variety of protein engineering applications have been reported in the literature. These applications range from biocatalysis for food and industry to environmental, medical and nanobiotechnology applications. Successful combinations of various protein engineering methods had led to successful results in food industries and have created a scope to maintain the quality of finished product after processing.

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Heterologous expression and protein engineering of wheat gluten proteins

Cited by 21 publications

References 96 publications

Effects of incorporated amaranth albumins on the functional properties of wheat dough

Effects of incorporated amaranth albumins on the functional properties of wheat dough

High-level expression of LMW-GS and α-gliadin genes promoted by the expressed tag sequence of 5′ end in Escherichia coli

Protein engineering and its applications in food industry

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