Enhanced production of recombinant protein inEscherichia coli using silkworm hemolymph

Kim, Ji Eun; Kim, Eun Jeong; Rhee, Won Jong; Park, Tai Hyun

doi:10.1007/bf02931854

Cited by 16 publications

(2 citation statements)

References 17 publications

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“…The study found that adding silkworm hemolymph to the culture medium can significantly inhibit the apoptosis of insect cells. The results confirmed the 30K protein in hemolymph shows antiapoptotic activity (J. E. Kim et al, 2005; Ying et al, 2013). The silkworm 30kc6 protein is one of the members of the 30K protein family, which greatly enhances cell viability, reduces DNA fragmentation, and inhibits the generation of cell reactive oxygen species (Yu et al, 2013).…”

Section: Introductionsupporting

confidence: 74%

Role of Bm30kc6 gene in cell apoptosis and the silk gland degradation signaling pathway in Bombyx mori L.

Xiao

Bibi

et al. 2020

Arch Insect Biochem Physiol

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Apoptosis is a process of programmed cell death that is regulated by genes independently. The Bm30kc6 gene is a kind of small molecular lipoprotein about 30 kDa, expressed highly in the late stage of the silkworm hemolymph. Our study showed that overexpression of Bm30kc6 could decrease caspase‐3 activation. Meanwhile, activation of caspase‐3 increased when Bm30kc6 expression was disturbed by small interfering RNA (siRNA). Cell apoptosis was decreased when Bm30kc6 was overexpressed under UV treatment. The apoptosis rate induced by actinomycin D is similar to the trend by UV. It was inferred that Bm30kc6 has an inhibitory effect on the apoptosis of silkworm cells. The apoptosis‐related genes, such as BmFadd, BmDredd, and BmDaxx were increased after overexpression of Bm30kc6 or decreased after interference of siRNA. It was speculated that there was an interactive relationship between Bm30kc6, BmDaxx, BmFadd, and BmDredd in the apoptosis signaling pathways. We investigated the transcription expression of the Bm30kc6 gene in different growth stages and tissues of the silkworm. The results showed that Bm30kc6 reached its peak in the hemolymph during the 6th to 7th days of the 5th instar, or in spinning post 24 h of the silk gland. In the silkworm BmN cells treated with caspase‐3/7 inhibitor, the caspase‐3 enzyme activity, and the expression levels of Bm30kc6, BmFadd, BmDredd, and BmDaxx were significantly reduced. The expression levels of Bm30kc6 increased sharply when silkworms were treated by molting hormone at Day 3 or 5 of the 5th instar. The results indicated that the expression of the Bm30kc6 gene was affected by the molting hormone and was likely to be its downstream target. In conclusion, the results suggest that the Bm30kc6 gene is involved in the regulation of the apoptotic signaling pathway and plays a role in the apoptotic process.

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Section: Introductionsupporting

confidence: 74%

Role of Bm30kc6 gene in cell apoptosis and the silk gland degradation signaling pathway in Bombyx mori L.

Xiao

Bibi

et al. 2020

Arch Insect Biochem Physiol

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“…It has been determined that the 30 K protein group in the SH, which consists of five proteins, constitutes a primary factor with regard to this inhibitory effect on apoptosis. SH was also known to enhance the production of recombinant protein [19]. SH was also known to enhance the production of recombinant protein [19].…”

Section: Fkqolar`qflk=mentioning

confidence: 99%

Effects of silkworm hemolymph and cartilage-specific extracellular matrices on chondrocytes and periosteum-derived progenitor cells

Shin

Choi

Lim

et al. 2006

Biotechnol. Bioprocess Eng.

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Enhancement of Wound Healing Efficacy by Increasing the Stability and Skin‐Penetrating Property of bFGF Using 30Kc19α‐Based Fusion Protein

Lee

Kim

et al. 2021

Advanced Biology

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involvement in tissue growth and regeneration, including wound healing. Activation of FGF receptor by bFGF stimulates intracellular signaling pathways such as phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling pathways which are related to the mitogenic response of cells. [1] As bFGF acts on various target cells involved in wound healing, its effects on wound healing have been extensively studied. [2] bFGF promotes the migration of dermal fibroblasts, keratinocytes, and endothelial cells, into wound sites and facilitates their proliferation. [3] As a result, bFGF accelerates granulation tissue formation and angiogenesis, which results in matrix formation and remodeling. [4] In recent years, a number of clinical uses of recombinant bFGF have been reported, and these include treatment of chronic wounds, second-degree burns, pressure ulcers, and diabetic foot ulcers. [5] However, its inherent instability and rapid degradation often require frequent treatments in tandem with higher concentrations. [6] The reported half-life of recombinant bFGF is less than 10 h under normal cell culture conditions and even shorter in the living organisms, which significantly limits The instability of recombinant basic fibroblast growth factor (bFGF) is a major disadvantage for its therapeutic use and means frequent applications to cells or tissues are required for sustained effects. Originating from silkworm hemolymph, 30Kc19α is a cell-penetrating protein that also has protein stabilization properties. Herein, it is investigated whether fusing 30Kc19α to bFGF can enhance the stability and skin penetration properties of bFGF, which may consequently increase its therapeutic efficacy. The fusion of 30Kc19α to bFGF protein increases protein stability, as confirmed by ELISA. 30Kc19α-bFGF also retains the biological activity of bFGF as it facilitates the migration and proliferation of fibroblasts and angiogenesis of endothelial cells. It is discovered that 30Kc19α can improve the transdermal delivery of a small molecular fluorophore through the skin of hairless mice. Importantly, it increases the accumulation of bFGF and further facilitates its translocation into the skin through follicular routes. Finally, when applied to a skin wound model in vivo, 30Kc19α-bFGF penetrates the dermis layer effectively, which promotes cell proliferation, tissue granulation, angiogenesis, and tissue remodeling. Consequently, the findings suggest that 30Kc19α improves the therapeutic functionalities of bFGF, and would be useful as a protein stabilizer and/or a delivery vehicle in therapeutic applications.

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Enhanced production of recombinant protein inEscherichia coli using silkworm hemolymph

Cited by 16 publications

References 17 publications

Role of Bm30kc6 gene in cell apoptosis and the silk gland degradation signaling pathway in Bombyx mori L.

Role of Bm30kc6 gene in cell apoptosis and the silk gland degradation signaling pathway in Bombyx mori L.

Effects of silkworm hemolymph and cartilage-specific extracellular matrices on chondrocytes and periosteum-derived progenitor cells

Enhancement of Wound Healing Efficacy by Increasing the Stability and Skin‐Penetrating Property of bFGF Using 30Kc19α‐Based Fusion Protein

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