Epidermal growth factor (EGF) is a representative ingredient in the cosmetics industry and has the ability to induce cell growth and proliferation. Recently, plant-based systems for producing high-value animal proteins such as EGF have garnered increasing attention in the field of biotechnology, owing to their low production cost, and the absence of endotoxins and animal viruses. However, the commercial viability of these recombinant proteins depends not only on their biological function but also on the efficiency and scalability of their production, particularly the achievable yield. In this study, we investigated whether fusing EGF with a macromolecule transduction domain (MTD), a type of cell-penetrating peptide, affects the production yield of EGF in plants. By comparing the expression levels of EGF with and without the MTD fusion, we aimed to evaluate the potential of MTD as a tool to enhance the production efficiency of functional proteins for commercial application.Seed-specific codon-optimized genes encoding human EGF with (ShEGF-MTD151) or without MTD (ShEGF) were transformed into soybean to compare the amount of their EGF proteins. Total 24 transgenic plants were produced, and T1 seeds were harvested from each soybean plant. The ShEGF and Bar transgenes were expressed in 17 lines (#2, #4–#10, #13–#16, #19–#21, #23, and #24) from pCKLSL:ShEGF, and 21 transgenic lines (#1–#3, #5–#10, #12–#19, and #21–#24) from pCKLSL:ShEGF-MTD151 expressed both ShEGF-MTD151 and Bar transgenes. To compare the level of EGF protein in pCKLSL:ShEGF and pCKLSL:ShEGF-MTD151 soybean T2 seeds, each seed sample was analyzed using an Enzyme-Linked Immunosorbent Assay (ELISA) kit for EGF quantification. The pCKLSL:ShEGF line #2 produced of 5.45 ng/g dry seed weight, whereas the pCKLSL:ShEGF-MTD151 line #16 showed 71.5 ng/g dry seed weight. pCKLSL:ShEGF-MTD151 (#16) T2 soybean seeds had a protein content 13 times higher than that of pCKLSL:ShEGF (#2) T2 soybean seeds. According to these results, fusion of an MTD into EGF could positively affect the production of EGF in soybeans. We assumed that these additional sequences may have a positive effect on translation, either by slowing down the degradation of EGF mRNA or by increasing mRNA stability. This work also demonstrates the feasibility of using soybean as a platform for producing bioactive human proteins and highlights the potential of MTD fusion technology to improve yield, thereby advancing plant-based bioproduction systems for industrial applications.

Comparison of Protein Production of Human Epidermal Growth Factor with or Without Macromolecule Transduction Domain in Transgenic Soybean Seed

期刊论文
发表日期：

2025年7月14日

查看全文

Hyeon Jin Song,
Hye Jeong Kim,
Si Hyeon Kim,
Wan Woo Yeom,
Young-Soo Chung

摘要

Epidermal growth factor (EGF) is a representative ingredient in the cosmetics industry and has the ability to induce cell growth and proliferation. Recently, plant-based systems for producing high-value animal proteins such as EGF have garnered increasing attention in the field of biotechnology, owing to their low production cost, and the absence of endotoxins and animal viruses. However, the commercial viability of these recombinant proteins depends not only on their biological function but also on the efficiency and scalability of their production, particularly the achievable yield. In this study, we investigated whether fusing EGF with a macromolecule transduction domain (MTD), a type of cell-penetrating peptide, affects the production yield of EGF in plants. By comparing the expression levels of EGF with and without the MTD fusion, we aimed to evaluate the potential of MTD as a tool to enhance the production efficiency of functional proteins for commercial application.

Seed-specific codon-optimized genes encoding human EGF with (ShEGF-MTD₁₅₁) or without MTD (ShEGF) were transformed into soybean to compare the amount of their EGF proteins. Total 24 transgenic plants were produced, and T₁ seeds were harvested from each soybean plant. The ShEGF and Bar transgenes were expressed in 17 lines (#2, #4–#10, #13–#16, #19–#21, #23, and #24) from pCKLSL:ShEGF, and 21 transgenic lines (#1–#3, #5–#10, #12–#19, and #21–#24) from pCKLSL:ShEGF-MTD₁₅₁ expressed both ShEGF-MTD₁₅₁ and Bar transgenes. To compare the level of EGF protein in pCKLSL:ShEGF and pCKLSL:ShEGF-MTD₁₅₁ soybean T₂ seeds, each seed sample was analyzed using an Enzyme-Linked Immunosorbent Assay (ELISA) kit for EGF quantification. The pCKLSL:ShEGF line #2 produced of 5.45 ng/g dry seed weight, whereas the pCKLSL:ShEGF-MTD₁₅₁ line #16 showed 71.5 ng/g dry seed weight. pCKLSL:ShEGF-MTD₁₅₁ (#16) T₂ soybean seeds had a protein content 13 times higher than that of pCKLSL:ShEGF (#2) T₂ soybean seeds. According to these results, fusion of an MTD into EGF could positively affect the production of EGF in soybeans. We assumed that these additional sequences may have a positive effect on translation, either by slowing down the degradation of EGF mRNA or by increasing mRNA stability. This work also demonstrates the feasibility of using soybean as a platform for producing bioactive human proteins and highlights the potential of MTD fusion technology to improve yield, thereby advancing plant-based bioproduction systems for industrial applications.