Co-expressing Turnip Crinkle Virus-coat protein with the serine protease α-thrombin precursor (pFIIa) in Nicotiana benthamiana Domin

Becher, Melina Laguía; Zaldúa, Zurima; Xu, Weijie; Marconi, Patricia Laura; Velander, William H.; Álvarez, María Alejandra

doi:10.1007/s11627-018-09956-0

Cited by 3 publications

(2 citation statements)

References 62 publications

(57 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The AmyS expression cassette is composed of a seed-specific promoter, the coding sequence for the Arabidopsis 2S albumin (At2S) signal peptide and AmyS, and the Arabidopsis heat shock protein 18.2 (AtHSP) terminator. The At2S signal peptide is anticipated to direct the synthesized protein to the endoplasmic reticulum [ 21 , 22 , 23 ]. The Nicotiana tabacum Rb7 matrix attachment region (MAR) was added to the 3’ end of the AmyS expression cassette.…”

Section: Resultsmentioning

confidence: 99%

Transgenic Soybean for Production of Thermostable α-Amylase

Cao,

Jiang,

et al. 2024

Plants

View full text Add to dashboard Cite

Alpha-amylases are crucial hydrolase enzymes which have been widely used in food, feed, fermentation, and pharmaceutical industries. Methods for low-cost production of α-amylases are highly desirable. Soybean seed, functioning as a bioreactor, offers an excellent platform for the mass production of recombinant proteins for its ability to synthesize substantial quantities of proteins. In this study, we generated and characterized transgenic soybeans expressing the α-amylase AmyS from Bacillus stearothermophilus. The α-amylase expression cassettes were constructed for seed specific expression by utilizing the promoters of three different soybean storage peptides and transformed into soybean via Agrobacterium-mediated transformation. The event with the highest amylase activity reached 601 U/mg of seed flour (one unit is defined as the amount of enzyme that generates 1 micromole reducing ends per min from starch at 65 °C in pH 5.5 sodium acetate buffer). The optimum pH, optimum temperature, and the enzymatic kinetics of the soybean expressed enzyme are similar to that of the E. coli expressed enzyme. However, the soybean expressed α-amylase is glycosylated, exhibiting enhanced thermostability and storage stability. Soybean AmyS retains over 80% activity after 100 min at 75 °C, and the transgenic seeds exhibit no significant activity loss after one year of storage at room temperature. The accumulated AmyS in the transgenic seeds represents approximately 15% of the total seed protein, or about 4% of the dry seed weight. The specific activity of the transgenic soybean seed flour is comparable to many commercial α-amylase enzyme products in current markets, suggesting that the soybean flour may be directly used for various applications without the need for extraction and purification.

show abstract

Section: Resultsmentioning

confidence: 99%

Transgenic Soybean for Production of Thermostable α-Amylase

Cao,

Jiang,

et al. 2024

Plants

View full text Add to dashboard Cite

show abstract

“…Furthermore, the entire FVIII factor was produced in the chloroplasts of lettuce ( Lactuca sativa ) at a level of 852 µg/g (Kwon et al 2018 ). Another blood coagulation factor, a serine protease a-thrombin precursor (pFIIa) involved in fibrin formation, was co-expressed in N. benthamiana with Turnip Crinkle Virus coat protein (TCV-CP), which is known to interfere with post-transcriptional gene silencing (Laguia-Becher et al 2019 ).…”

Section: Three Types Of Pmpsmentioning

confidence: 99%

Plant-made pharmaceuticals: exploring studies for the production of recombinant protein in plants and assessing challenges ahead

Lee

Park

et al. 2023

Plant Biotechnol Rep

View full text Add to dashboard Cite

The production of pharmaceutical compounds in plants is attracting increasing attention, as plant-based systems can be less expensive, safer, and more scalable than mammalian, yeast, bacterial, and insect cell expression systems. Here, we review the history and current status of plant-made pharmaceuticals. Producing pharmaceuticals in plants requires pairing the appropriate plant species with suitable transformation technology. Pharmaceuticals have been produced in tobacco, cereals, legumes, fruits, and vegetables via nuclear transformation, chloroplast transformation, transient expression, and transformation of suspension cell cultures. Despite this wide range of species and methods used, most such efforts have involved the nuclear transformation of tobacco. Tobacco readily generates large amounts of biomass, easily accepts foreign genes, and is amenable to stable gene expression via nuclear transformation. Although vaccines, antibodies, and therapeutic proteins have been produced in plants, such pharmaceuticals are not readily utilized by humans due to differences in glycosylation, and few such compounds have been approved due to a lack of clinical data. In addition, achieving an adequate immune response using plant-made pharmaceuticals can be difficult due to low rates of production compared to other expression systems. Various technologies have recently been developed to help overcome these limitations; however, plant systems are expected to increasingly become widely used expression systems for recombinant protein production.

show abstract