• Journal of Life Science
• /
• v.23 no.1
• /
• pp.15-23
• /
• 2013

AprE51 from Bacillus amyloliquefaciens CH51 is a 27 kDa subtilisin-like protease with fibrinolytic activity. AprE51-6 showing increased catalytic activity was produced previously. To enhance the thermostability of AprE51-6, 2 residues, Gly-166 and Asn-218 based on B. subtilis subtilisin E were mutated by site-directed mutagenesis. The results of the mutational analysis showed that substitution of arginine for Gly-166 (AprE51-7) increased the fibrinolytic activity 1.8-fold. An N218S mutant (AprE51-8) also increased the fibrinolytic activity up to 4.5-fold in a fibrin plate assay. Purified AprE51-7 and AprE51-8 mutants had a 1.9- and a 2.5-fold higher $k_{cat}$, respectively, and a 2.1-1.9-fold lower $K_m$, respectively. This resulted in a 3.8- and a 4.7-fold increase in catalytic efficiency ($k_{cat}/K_m$), respectively, relative to that of wild-type AprE51. AprE51-8 had a broader pH range than AprE51-6 and nattokinase, especially at an alkaline pH value. In addition, AprE51-8 showed higher thermostability than AprE51-6 at $60^{\circ}C$. The half-lives of AprE51-7 and AprE51-8 at $50^{\circ}C$ were 21.5 and 27.3 min, respectively, which are 2.0 and 2.6 times longer, respectively, than that of the wild-type AprE51.

• Journal of Life Science
• /
• v.27 no.2
• /
• pp.123-130
• /
• 2017

This study was conducted to investigate the physicochemical quality characteristics of pork patty added with four different amount (T0:0%, T1:0.3%, T2:0.7%, T3:1.0%) of tangerine (Citrus unshiu) peel. There was no significant difference in chemical composition, cooking yield, water holding capacity, moisture retention, fat retention, hardness, springiness, cohesiveness, gumminess, chewiness, VBN content, L-value. In taste, texture, juiciness and palatability, the addition of 0.3%~1.0% tangerine peel in pork patty showed no significant difference on sensory properties compared to the pork patty without tangerine peel. Total polyphenol content was highest in T3, and DPPH radical scavenging activity was highest in T2 and T3 (p<0.001). The TBARS contents decreased as tangerine peel become added (p<0.001). The pH was highest in T0, and was lowest in T3 (p<0.001). The external a-value of T2 and T3 were significantly higher than that of T0 (p<0.01). The external and internal b-value of T2 and T3 were higher than those of T0 (p<0.01). Flavor of T2 and T3 were higher than those of T0 and T1 (p<0.01). In conclusion, the results of this study indicate that an addition of tangerine peel could be utilized as an ingredient in pork patty in promotion of function of tangerine by-products.

• Journal of Life Science
• /
• v.27 no.2
• /
• pp.202-210
• /
• 2017

As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

• Journal of Life Science
• /
• v.27 no.7
• /
• pp.739-745
• /
• 2017

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy. $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) predominantly converts progesterone into its biologically inactive form $20{\alpha}$-hydroxyprogesterone ($20{\alpha}$-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In this study, we characterized the expression and localization of $20{\alpha}$-HSDinthe testis of MediKinetics $Micropigs^{(R)}$. The testes were collected at days 6, 9, 12, 18, and 21 after birth. The $20{\alpha}$-HSD mRNA was found to be expressed in the testis at day 6 after birth by RT-PCR. The highest level of mRNA expression in the testis was detected on day 21 after birth. However, the mRNA was not detected in the placenta after parturition. Western blot for $20{\alpha}$-HSD reveal that the specific 37-kDa band was detected in immature pig testis. However, this band was not detected in testis tissue at day 6 after birth. In the immunohistochemical analysis of the testis, $20{\alpha}$-HSD was detected in the Sertoli cells and Leydig cells. Taken together, our study shows for the first time that the $20{\alpha}$-HSD mRNA and protein are expressed in pig testis after birth. Further investigation is required to elucidate the functional mechanisms of $20{\alpha}$-HSD in pig testis after birth.

• Journal of Life Science
• /
• v.27 no.11
• /
• pp.1256-1261
• /
• 2017

${\beta}$-glucan is a constituent of the cell wall of fungi, yeast and plants. It plays an important role in the immune system such as activation of immunocyte, release of pro-inflammatory and anti-cancer effect. The immune system maintains a healthy immune homeostasis. However, when pathogenic substances enter the body, immune homeostasis can break down and disease can be triggered. Therefore, we studied a substance that regulates immune homeostasis. The purpose of the study we demonstrated whether the ${\beta}$-glucan can be applied to the immune-modulation effects in human monocytic THP-1 cells. ${\beta}$-glucan was incubated in THP-1 cells at various concentrations. The $TNF-{\alpha}$ mRNA expression and protein levels were analyzed by ELISA and Real-time PCR. Additionally, the expression of MAPKs (p38 and JNK), $I{\kappa}B-{\alpha}$ and $NF-{\kappa}B$ p50 were analyzed by western blot. ${\beta}$-glucan enhanced the production of $TNF-{\alpha}$ mRNA expression and protein levels in human monocytic THP-1 cells. In addition, activation of MAPKs (p38 and JNK) and $NF-{\kappa}B$ p50 induced by ${\beta}$-glucan were increased. The study suggests that ${\beta}$-glucan contributes to immune-stimulation effect by production $TNF-{\alpha}$ in human monocytic THP-1 cells, and that MAPKs and $NF-{\kappa}B$ p50 are involved in the process. Synthetically, we have suggested ${\beta}$-glucan may be improved to immune system effect in human monocytic THP-1 cells.

• Journal of Life Science
• /
• v.28 no.5
• /
• pp.621-626
• /
• 2018

We introduced the physiological responses of aging, active aging and also suggest the impact of physical exercise on body health status and elderly immunity. In this purpose, we searched the Pub Med data base for the articles (include our experimental papers) and review papers having the terms 'Aging', 'Active aging' and 'Physical activity and elderly' in the title, published from 1999 until 2018. The results were as follows: Exercise training has been extensively studied about the reduction of inflammation, oxidative stress, disease, and aging in syndrome X patients and elderly. Combined and aerobic or resistance exercise training could reduce obesity, insulin resistance, type 2 diabetes and hypertension. Exercise training has been extensively studied in cancer settings as part of prevention or treatment strategies. From this research, regular exercise has the potential to target tumor growth through regulation of inflammation and immune responses such as lactate clearance, NK cell activation (innate immunity), activation of cytotoxic immune cells, T cell activation (adaptive immunity), and immune surveillance. However, Endurance physical activity not only induces thermogenesis and diverse sports injuries but also elicits mobilization and functional enhancement of monocytes, neutrophils (which is caused by the cytokine changes such as TNF-alpha, IL-1) whereas it suppresses cell mediated immunity causing to increased susceptibility to inflammation and infections like cough and URTIs (upper respiratory track infections) in young and especially in elderly people. Therefore, Strategies to prevent physical fatigue, sports injuries include avoid overtraining, Adequate recovery and various type of rest during and after physical activity and assuring adequate nutrition supplementation such as glutamine, vitamin B, vitamin C, carbohydrate, ion or berry-contain sports beverages is helpful in physically active elderly.

• Journal of Life Science
• /
• v.25 no.7
• /
• pp.740-747
• /
• 2015

This study was carried out to evaluate changes in the meat quality and antioxidation activity in the loin and ham of Korean Native Black Pigs (KNBP) during frozen storage at −18℃ for 150 days. The pH value of the loin was decreased as storage days progressed, while the pH value of the ham showed no consistent changes with storage days. The lightness (L*) of the loin did not show any significant reduction until day 120, whereas L* of the ham was significantly declined throughout the storage period (p<0.05). The redness (a*) values of the loin and ham were significantly decreased as storage progressed. The water holding capacity of the loin was decreased by day 30 and that value was maintained until the end of storage. The initial total numbers of microorganisms in the loin and ham were 4.88 and 5.16 Log CFU/g, respectively and these numbers were significantly decreased by day 30 (p<0.05). The levels of 2-thiobarbituric acid reactive substances (a measurement of lipid oxidation) in the loin and ham ranged from 0.057-0.069 and 0.052-0.087 mg MDA/kg meat, respectively, until storage day 150. Volatile basic nitrogen values of the loin and ham ranged from 15.13-16.55 and 16.05-16.23 mg%. Oxygen radical absorbance capacities and carnosine contents of the loin and ham were significantly decreased during frozen storage for 3 months (p<0.05). In summary, the meat quality of the loin and ham from KNBP was somewhat decreased during frozen storage. However, the levels of antioxidants and dipeptides with antioxidant activity were significantly decreased in pork loin and ham during frozen storage.

• Journal of Life Science
• /
• v.19 no.11
• /
• pp.1611-1616
• /
• 2009

This study was conducted to determine the characteristics in breeding of the little tern Sterna albifrons in Shinja-do and Doyo-Deung in Nakdong estuary. The data was obtained in the middle and late 1990's and in the mid 2000's, which were analysed for the breeding information of the little tern. In Shinja-Do, a total of 638 nests were observed in 1995, and the average numbers of eggs laid per nest were 2.48. In 2004 and 2005, 94 and 456 nests were observed, and the average number of eggs laid per nest was 2.16 and 2.53, respectively. There was a significant difference between the average numbers of eggs laid per nest for the 3 respective years. In Doyo-Deung, a total of 269 nests were observed in 1998, and the average number of eggs laid per nest was 2.16. In 2004 and 2005, 1,023 and 1,123 nests were observed, and the average numbers of eggs laid per nest were 2.33 and 2.52, respectively. There was a significant difference (p<0.001) between the average number of eggs laid per nest as well. Hatching success rate by nesting site in Shinja-Do was calculated in 1995. Type A nest (nest located on dry sand land) showed a 56.8% success rate, and type B nest (nest located on the land with dwarf grass), type C nest (nest located on moist land) and type D nest (nest located on the land with shells) showed 55.3%, 36.6% and 59.3% success rates, respectively. The average hatching success rate in Shinja-Do was 54.5%. The most crucial factor for hatching failure was flood, especially, for the type C nests. The hatching success rate by the nesting site in Doyo-Deung showed an average of 73.2%. It was high in type D nest (83.8%) and type A nest (75.9%), and low in type C nest (59.7%). Again, the most crucial factor for hatching failure was flood, especially, for the type C nests. Nakdong estuary has been through many changes over time, particularly in recent. Therefore, continuous efforts to keep it a breeding place for the little tern are urgently required, including overall management of land, protection of reef and sand, etc.

• Journal of Life Science
• /
• v.20 no.3
• /
• pp.457-461
• /
• 2010

Familial hypokalemic periodic paralysis (HOKPP) is an autosomal dominant disorder characterized by reversible flaccid paralysis and intermittent hypokalemia. Although it has been reported that decreased activity in the $K_{ATP}$ channels of the skeletal muscle cell membrane plays a role in the pathogenesis of HOKPP, a clear mechanism has not yet been established. This study aimed to investigate the molecular biological mechanism underlying the decreased activity of $K_{ATP}$ channels in the skeletal muscles of familial HOKPP patients by studying the levels of the $K_{ATP}$ channel subunit Kir6.2. We found that when cells obtained from healthy individuals (normal cells) and HOKPP patients (patient cells) were treated with 4 mM potassium buffer, there was no quantitative change in the KCNJ11 mRNA levels and no difference in the Kir6.2 protein expression in the cytosol and cell membrane. On the other hand, when 1 mM potassium buffer was used, normal cells showed decreased expression of KCNJ11 mRNA as well as decreased expression of Kir6.2 protein in the cell membrane. However, patient cells treated with the same buffer showed no quantitative change in the levels of KCNJ11 mRNA or in the levels of Kir6.2 protein in the cytosol and cell membrane. Thus, in HOKPP patients, the Kir6.2 protein cannot be transported from the cell membrane to the cytosol, leading to closure of the $K_{ATP}$ channels, induction of depolarization, and subsequently, to the paralytic symptoms observed in the patient. Our findings thus provide new insights into the pathogenesis of HOKPP.

• Journal of Life Science
• /
• v.19 no.8
• /
• pp.1039-1046
• /
• 2009

The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.