• Biotechnology and Bioprocess Engineering:BBE
• /
• v.10 no.4
• /
• pp.353-356
• /
• 2005

The effect of silkworm hemolymph on the expression of recombinant protein in Escherichia coli was investigated. The addition of silkworm hemolymph to the culture medium in­creased the production of recombinant $\beta$-galactosidase in E. coli. The production was dependent on the concentration of the added silkworm hemolymph, which increased 2-, 5-, and 8-fold in media supplemented with 1, 3, and $5\%$ silkworm hemolymph, respectively. To identify the effective component, the silkworm hemolymph was fractionated by gel filtration column chromatography. A fraction, with a molecular weight of about 30 K was identified as the effective component.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.5
• /
• pp.853-857
• /
• 2001

The effect of silkworm hemolymph on the inhibition of baculovirus-induced insect cell apoptosis was quantitatively investigated using a flow cytometric analysis. Spodoptera frugiperda (Sf9) cell and Autographa californica nuclear polyhedrous virus (AcNPV) were used as a model for insect cell and baculovirus in this study, respectively. Compared with a mammalian cell cycle, the fraction of G1 cells was relatively small in the Sf9 cell cycle. Silkworm hemolymph did not affect the Sf9 cell-cycle distribution before the baculovirus infection. However, the fraction of cells which are not in the sub-G1 phase remained at a high level for 3 days after the infection in the medium without silkworm hemolymph, while it remained at a high level for 7 days after the infection in the medium supplemented with silkworm hemolymph. The fractions of apoptotic cells in the sub-G1 phase were $4.7\%$, and 4 days after infection, $22.7\%$, in the media with and without silkworm hemolymph, respectively.

• Journal of Radiation Industry
• /
• v.8 no.1
• /
• pp.7-10
• /
• 2014

This study was designed to evaluate the protective effect of silkworm hemolymph against ${\gamma}-irradiation$ induced damage in the liver of mice. Female Balb/C mice (6 weeks old) were exposed to ${\gamma}-irradiation$ (6 Gy) and administered orally to silkworm hemolymph ($5ml\;kg^{-1}$ BW) for 7 days post-irradiation. The body weight, spleen index, plasma aspartate transaminase (AST), plasma alanine transaminase (ALT), and liver malondialdehyde (MDA) levels were determined. Compared with irradiated control mice, the activity of plasma AST and the level of MDA were significantly decreased in mice treated silkworm hemolymph. These results show that silkworm hemolymph is found to have a protective effect against ${\gamma}-irradiation$ induced damage in mice.

• 한국생물공학회:학술대회논문집
• /
• 2000.11a
• /
• pp.151-154
• /
• 2000

The effect of silkworm hemolymph on insect cell apoptosis was investigated. The addition of silkworm hemolymph into the culture medium either before or during the baculovirus infection increased the host cell longevity. Silkworm hemolymph also inhibits apoptosis induced by actinomycin D, the RNA synthesis inhibitor. In this study, a flow cytometry was used for the quantitative analysis of apoptosis inhibition by silkworm hemolymph.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.3 no.2
• /
• pp.87-90
• /
• 1998

Hemolymph is oxidized and darkens visibly during the collection from silkworm due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation ; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. It makes the hemolymph collection difficult especially on a large-scale preparation. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as Lemone, TBO, Cre, Y4, and wEb showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The hemolymph of wEb showed the slowest oxidation. The absorbance of this mutant hemolymph reached the saturation value at 20$^{\circ}C$ in 450 min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. We tested if this mutant hemolymph is useful as a medium supplement for insect cell culture. Cell growth rate and final cell concentration in the medium supplemented with the wEb hemolymph were almost same as those in the medium supplemented with the wild-type hemolymph. Hemolymph is collected on a small scale by clipping the abdominal leg; however, this method is not appropriate fro large scale preparation. Centrifugation after chopping the silkworm hemolymph by a blending mixer is a more appropriate procedure for large scale collection. Slowly oxidized wEb hemolymph resulted in higher cell concentration than the wild-type hemolymph when hemolymph was collected by the large scale preparation method.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.5 no.2
• /
• pp.189-194
• /
• 2002

Silkworm diseases are major constraint in silk cocoon production. Among silkworm diseases, white muscar-dine is highly contagious and most common in winter and rainy seasons. It is suggested that hemocytes involve in defense against invasion of Beauveria bassi-ana and systemic fungicide/chemicals prevent the proliferation of fungi in the hemolymph or preventing the growth of the fungi in the body cavity through enhancing the hemocyte mediated defense response. In the present study the influence of systemic fungicide on hematological changes in silkworms infected with Beauveria bassiana was reported. It is observed that the total hemocyte counts increased in the hemolymph up to 5 days post inoculation in systemic fungicide treated batches while in the inoculated control the increase was up to 3 days indicating the positive hemocyte mediated response in silkworm treated with systemic fungicide. After 2 days in the inoculated control as the multiplication and growth of mycelia increased, defense capacity of the silkworm was decreased. The biochemical changes were also observed in the hemolymph of silkworm infected with B. bassiana. In silkworm infected with Beauveria bassi-ana, the total protein content increased whereas total carbohydrate and total lipids decreased as the infection progresses. In the case of systemic fungicide treated batches the increase in total protein content was comparatively higher and decrease in total carbohydrate and lipids were comparatively lower than the inoculated control.

• Journal of Sericultural and Entomological Science
• /
• v.37 no.2
• /
• pp.181-185
• /
• 1995

To enhance expression of foreign gene by the novel expression vector, pBmKSK1, of Bombyx mori nuclear polyhedrosis virus, E. coli $\beta$-galactosidase gene expressing recombinant virus was infected in BmN-4 cells and various concentrations of silkworm hemolymph were added to the recombinant virus-infected BmN-4 cells containing fetal bovine serum. The expression efficiency of foreign gene was determined by $\beta$-galactosidase activity in the culture media. The results showed that the silkworm hemolymph was effective to expression of foreign gene in the BmN-4 cells, suggesting that the silkworm hemolymph could be substituted for fetal bovine serum in the BmN-4 cells to enhance expression of foreign gene.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.2
• /
• pp.227-229
• /
• 1999

Animal cell culture media are usually supplemented with fetal bovine serum (FBS); however, the use of FBS presents certain problems including high cost. By using an adaptation process and the addition of silkworm hemolymph, the FBS concentration can be reduced without causing a significant decrease in cell growth.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.8 no.1
• /
• pp.117-121
• /
• 2004

Soluble proteins of silkworm hemolymph were investigated by means of isoelectric focusing (IEF). The protein spectra during ontogenesis of races and inter-races hybrids kept in Bulgaria was studied. A total of 51 protein bands in the hemolymph from fourth larval instar to imago were ascertained. Stage specific expression was established. The specific expression of some protein bands in the individual spectra manifest phenotype of gene determinate polymorphism (HP F, HP J, HP K, HP L, HP Q - in the zone with pH gradient 3.5-6.2 and HP K, HP L, HP N, HP P, HP T - in the zone with pH gradient 9.5 - 6.2). Breed specific expression was observed. On the basis of the obtained results, it was established that the investigated breeds are heterogeneous and the isoelectric focusing method is successful when specifying the inner-race and inter-race polymorphism in silkworm.

• Journal of Sericultural and Entomological Science
• /
• v.51 no.2
• /
• pp.207-210
• /
• 2013

In this study, matured silkworm hemolymph was studied for inhibitory effect on melanin formation, tyrosinase activities. A matured silkworm hemolymph mass purification method was developed and some of suppressing melanin formation effects were investigated, too. As a result, the advanced purification method was higher in yield rate than existing one by 53%. A free radical scavenging activity of hemolymph was higher than sericin on the market($IC_{50}$, $202.9{\pm}33.1$) and lower than vitamin C($0.57{\pm}0.02$) by $6.4{\pm}0.3{\mu}g/m{\ell}$. An inhibitory effect on melanin synthesis was higher in hemolymph compared to Morus bark and arbutin by 9.15% and 11.56%, respectively. An inhibitory effect on tyrosinase activity was higher in hemolymph than Morus bark and lower than arbutin. Effect of hemolymph $45{\mu}g/m{\ell}$ on suppressing of tyrosinase was almost 90%. Thus, these results suggest that hemolymph of matured silkworm may have beneficial properties as a material for cosmetics.