• International Journal of Industrial Entomology and Biomaterials
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• v.48 no.3
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• pp.107-113
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• 2024

The efficiency of protein extraction from Hongjam, a steamed mature silkworm, was quantitatively evaluated using various chemical buffers and physical methods. This study considers the difficulty of protein extraction yield due to the high content of hydrophobic amino acids in Hongjam compared to 5^th instar-3^rd day silkworm larvae. Results indicated that urea buffer enhanced protein yield more effectively than RIPA buffer. Additionally, the application of physical methods such as microwave treatment to samples treated with RIPA buffer increased yields by up to 22%, achieving concentrations as high as 3.9 mg/mL. Circular dichroism (CD) analysis showed that proteins extracted with urea buffer retained their structural integrity, exhibiting deeper and more prominent peaks associated with random coil structure. In addition, physical methods such as vortexing, sonication, microwave and homogenization increased the extraction yield of larger molecules without altering protein structures, suggesting their potential scalability for industrial applications. These results demonstrate the critical role of selecting appropriate extraction methods to optimize the yield and functionality of proteins from Hongjam, with implications for its use in biotechnological applications and nutraceuticals.

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.219-223
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• 2012

The present study was performed to identify the relationship between plasminogen activator (PA) and Heat Shock Protein-90 (HSP-90) in porcine uterus tissues during the estrous cycle. Porcine uterus tissues were obtained from preovulatory (Pre-Ov), post-ovulatory (Post-Ov) and early to mid-luteal (Early-mid L) stages. The protein was extracted from uterus tissue by using M-PER Mammalian Protein Extraction Reagent. Proteins were refined by RIPA Buffer and quantified by BCA methods. As results, t-PA expression was significantly (p<0.05) higher from pre-ovulatory(Epithelium tissue: $29,067{\mu}g/{\mu}l$, Myometrium tissue: $30,797{\mu}g/{\mu}l$) compared to the post-ovulatory stage(Epithelium tissue: $54,357{\mu}g/{\mu}l$, Myometrium tissue: $53,270{\mu}g/{\mu}l$) and early to mid-luteal stage(Epithelium tissue: $42,380{\mu}g/{\mu}l$, Myometrium tissue: $43,139{\mu}g/{\mu}l$). On the other hand, the uPA expression indicated higher from early to mid-luteal stage (Epithelium tissue: $0.02198{\mu}g/{\mu}l$, Myometrium tissue: $0.02412{\mu}g/{\mu}l$) than pre-ovulatory stage (Epithelium tissue: $0.01577{\mu}g/{\mu}l$, Myometrium tissue: $0.01531{\mu}g/{\mu}l$) and post-ovulatory stage(Epithelium tissue: $0.01414{\mu}g/{\mu}l$, Myometrium tissue: $0.01429{\mu}g/{\mu}l$). However, expression of u-PA did not differ from each estrous cycle in the epithelium tissue and myometrium tissue(p<0.05). Expression of HSP-90 was differ t-PA and u-PA from pre-ovulatory in Epithelium tissue($25,423{\mu}g/{\mu}l$) and early to mid-luteal stage in epithelium tissue($177,922{\mu}g/{\mu}l$) and myometrium tissue($26,664{\mu}g/{\mu}l$). These results suggest that HSP-90 and u-PA were related with change of uterus cycle according to the reformation of the tissues in porcine uterus.