Abstract : Keratin, which is the hard and insoluble structural protein found in many parts of birds, reptiles, and tissues such as feathers, hair, horns, wool. Some keratinaceous materials such as feathers known as industrial wastes, hard to degrade naturally because of their complex structure. To date, several bacteria and fungi have been discovered that have keratinase activity, which breaks down keratin into amino acids. In addition, various studies are being conducted to increase the activity of this enzyme through of recombinant keratinase or to facilitate purification. This potential enzyme can be used not only for the degradation of keratinaceous industrial wastes but also for the leather and healthcare industries. In this review, keratin degradation mechanisms, keratinase production, and their industrial applications will be discussed.
Abstract : Essential oils extracted from plants have been attracted significant attention in recent years due to their pharmacological uses such as for sterilization and preservation. In this study, essential oil was extracted from needles, twigs, and barks of pine trees using three different types of solvents: acetone, petroleum ether, and toluene. The oil extraction ratio of these three different parts of pines trees by each type of solvent was investigated in detail. In addition, TLC and FTIR analysis were performed to analyze and identify the essential oil extracted from the pine trees. When acetone was used for the solvent, the extraction ratio showed the highest value with the needles and decreased in the following order: needles > twigs > barks. In contrast, petroleum ether and toluene resulted in the highest extraction ratio with the twigs followed by the needles and barks. Although the boiling points of acetone and petroleum ether are similar, the average oil extraction ratio of the pine tree by acetone showed 4.3 times higher than petroleum ether attributed to the high polarity of acetone.
Abstract : Chinese hamster ovary (CHO) cells have been used for the production of therapeutic recombinant proteins, including monoclonal antibodies. CHO cell lines producing recombinant protein are developed traditionally by random integration, leading to clonal variation issues, arbitrarily various phenotypic characteristics between clones. Thus, development of inexpensive and fast strategy for transgene integration site analysis is necessary. In this study, we optimized Alu polymerase chain reaction (Alu PCR) for CHO cells using short interspersed nuclear elements (SINEs) in the genome. Uracilcontaining primers were used to specifically amplify the target between SINE and recombinant protein coding transgene. We applied this strategy to identify transgene integration site as well as the endogenous housekeeping gene as a control. In conclusion, we verify that Alu PCR can be an inexpensive and rapid alternative analysis tool to detect integration sites.
Abstract : Barley Sprout is a young sprout of barley and contains various nutrients with antioxidant effects. It also has various physiological effects including antioxidant, anti-obesity, and anti-diabetic effects. This study investigated the immunity-enhancing effect of Hot Water extract of Barley Sprout (BSE). According to the MTT assay results, BSE was not cytotoxic to RAW 264.7 macrophage cell line. Therefore, subsequent experiments, including the NO (nitric oxide) assay, Western blot assay, and WST-1 assay, were then conducted. First, to confirm how BSE affects the production of NO and Cytokine expression, which are biomarkers of immune cell activity, we treated the cells with LPS (lipopolysaccharide), which served as the positive control, and BSE at varying concentrations. The results show that BSE increased the production of NO and Cytokines, such as TNF-α (Tumor necrosis factor-alpha), IL-6 (Interleukin-6) and IL-1β (Interleukin-1beta). Additionally, the proliferation of Molt-4 cell line (T-cell) and Raji cell line (B-cell) increased in a concentration-dependent manner compared with the control. Taken together, these findings suggest that BSE consumption can improve immunity by enhancing innate immunity via activation of macrophages.
Abstract : Shake flasks are widely used as small-scale mammalian culture vessels during cell line development and bioprocess optimization for the production of biopharmaceuticals. However, the lack of information as well as controllability on dissolved oxygen often leads to oxygen limitation that can adversely affect cell growth and productivity during cell culture, thereby challenging cell line development and scale-up processes. In this work, we report an oxygen transfer rate (OTR) prediction model of shake flasks using Chinese hamster ovary (CHO) cells to avoid oxygen limitation issues in shake flasks. First, we varied shaking speed and culture volume to measure maximum OTR (OTRmax) and created an OTRmax regression function. Then, we estimated theoretical maximum viable cell density with the specific oxygen uptake rate (sOUR) of CHO cells and the OTRmax values. Lastly, we tested if decreasing culture volume or increasing shaker speed could mitigate the adverse effect of oxygen limitation. In conclusion, we believe this OTR prediction model would provide information about the oxygen availability in shake flasks, presumably assisting to achieve better culture performances.
Seong-hwan Yeom, Deok-Ho Kwon, Myoung-Dong Kim, Bong-Hwan Chung, and Suk-Jin Ha
Korean Society for Biotechnology and Bioengineering Journal 2021; 36(3): 192-197
Chae Hwan Cho, Ji-Hong Kim, Jae Hwan Shin, Hyo-Jung Yang, and Jong Pil Park*
Korean Society for Biotechnology and Bioengineering Journal 2021; 36(4): 221-230
Soo Hyun Park, Sangwon Chung, Jin-Taek Hwang, and Jae Ho Park
Korean Society for Biotechnology and Bioengineering Journal 2021; 36(3): 181-191
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