• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.6
• /
• pp.949-956
• /
• 2020

Objective: This study was conducted to confirm the effects of new inoculants producing-antifungal or esterase substances on rye silage and its rumen fermentation indices by comparing wild with mutated types. Methods: Rye harvested at dough stage was ensiled into 3 L mini bucket silo (1 kg) for 90 d in triplicate following: distilled water at 20 μL/g (CON); Lactobacillus brevis 100D8 (AT) and its inactivation of antifungal genes (AT-m) at 1.2×10⁵ cfu/g, respectively; and Leuconostoc holzapfelii 5H4 (FD) and its inactivation of esterase genes (FD-est) at 1.0×10⁵ cfu/g, respectively. After silo opened, silage was sub-sampled for the analysis of ensiling quality and its rumen fermentation indices. Results: Among the wild type inoculants (CON vs AT vs FD), FD inoculant had higher (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber, the total degradable fraction, and total volatile fatty acid in rumen, while AT inoculant had higher (p<0.05) lactate, acetate, and lactic acid bacteria in silage. Silage pH and the potentially degradable fraction in rumen increased (p<0.05) by inactivation of antifungal activity (AT vs AT-m), but lactate, acetate, and lactic acid bacteria of silage decreased (p<0.05). In silage, acetate increased (p<0.05) by inactivation of esterase activity (FD vs FD-est) with decreases (p<0.05) of pH, ammonia-N, lactate, and yeast. Moreover, inactivation of esterase activity clearly decreased (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber, the total degradable fraction, and total volatile fatty acid in the rumen. Conclusion: This study concluded that FD inoculant confirmed esterase activity on rye silage harvested at dough stage, while AT inoculant could not be confirmed with antifungal activity due to the absence of mold in all silages.

• The Korean Journal of Mycology
• /
• v.43 no.2
• /
• pp.99-103
• /
• 2015

We prepared supernatants and cell-free extracts of yeasts isolated from wild flowers in Korea, and their antioxidant activity and inhibitory activities of xanthine oxidase and tyrosinase were investigated. Among them, cell-free extracts of Kuraishia capsulata UL40-2 and Sporobolomyces ruberrimus 121-Z-3 showed significantly high xanthine oxidase inhibitory activity of 46.4% and 48.3%, respectively. Starmerella bombicola 80-J-1 also showed tyrosinase inhibitory activity of 36.2% in the cell-free extract. Other antioxidant activities and tyrosinase inhibitory activities were not detected or were below 20%. Maximal production of the xanthine oxidase inhibitors were observed when Kuraishia capsulate UL40-2 and Sporobolomyces ruberrimus 121-Z-3 were cultured in the yeast extract-peptone-dextrose media at $30^{\circ}C$ for 24 hour, respectively.

• Microbiology and Biotechnology Letters
• /
• v.42 no.4
• /
• pp.402-406
• /
• 2014

Six kinds of newly recorded yeasts such as Rhodosporidium diobovatum SY4-2, Cryptococcus bestiolae SY7-1, Kazachstania unispora SY14-1, Kazachstania servazzii SY14-3, Pichia holstii SY20-2 and Cryptococcus tephrensis SY26-1 were screened from sixty one yeasts derived from wild flowers found in Seonyudo, Gogunsanyeoldo, Jeollabuk-do, Korea. All of them grew in 50% glucose-containing yeast extract-peptone-dextrose (YPD) broth and Pichia holstii SY20-2 was also halophile, growing in 20% NaCl-containing YPD broth. All of them, except Cryptococcus tephrensis SY26-1, were assimilated to glucose. Cell-free extract from Kazachstania servazzii SY14-3 showed the highest 98.6% of ${\alpha}$-glucosidase inhibitory activity and maximal production of the ${\alpha}$-glucosidase inhibitor was obtained with 24h incubation at $30^{\circ}C$. The antihypertensive angiotensin I-converting enzyme inhibitory activity of the unrecorded yeasts were showed 58.6-80.4% in their supernatants.

• Bulletin of the Korean Chemical Society
• /
• v.35 no.9
• /
• pp.2781-2786
• /
• 2014

Human ${\alpha}_1$-antitrypsin (${\alpha}_1$-AT) is a natural inhibitor of neutrophil elastases and has several dozens of genetic variants. Most of the deficient genetic variants of human ${\alpha}_1$-AT are unstable and cause pulmonary emphysema. However, the most clinically significant variant, Z-type ${\alpha}_1$-AT, exhibits retarded protein folding that leads to accumulation of folding intermediates. These aggregate within the endoplasmic reticulum (ER) of hepatocytes, subsequently causing liver cirrhosis as well as emphysema. Here, we studied the role of an ER folding assistant protein Cpr5p on Z-type ${\alpha}_1$-AT folding. Cpr5p was induced > 2-fold in Z-type ${\alpha}_1$-AT-expressing yeast cells compared with the wild type. Knockout of CPR5 exacerbated cytotoxicity of Z-type ${\alpha}_1$-AT, and re-introduction of CPR5 rescued the knockout cells from aggravated cytotoxicity caused by the ${\alpha}_1$-AT variant. Furthermore, Cpr5p co-immunoprecipitated with Z-type ${\alpha}_1$-AT and facilitated its protein folding. Our results suggest that protein-folding diseases may be suppressed by folding assistant proteins at the site of causal protein biosynthesis.

• 한국생물공학회:학술대회논문집
• /
• 2003.10a
• /
• pp.753-757
• /
• 2003

Escherichia coli and Saccharomyces cerevisiae have been used as host for production of recombinant proteins. It is known that S. cerevisiae has advantages such as good folding and secretion capability, and safety as host over E. coli. But S. cerevisiae has shortcomings of low expression level which is just 20% of that of E. coli. To solve this problem, directed evolution method was tried to enhance the GAP promoter strength of S. cerevisiae in this study. As result, modified GAP promoter that has increased expression level of about 360% compared to that of wild type was selected.

• Korean Journal Plant Pathology
• /
• v.11 no.3
• /
• pp.265-270
• /
• 1995

Transposon mutation of Xanthomonas campestris pv. vesicatoria (Xcv) was induced by using transposon omegon ($\Omega$)-Km (Tn $\Omega$Km), which was confirmed by resistance to kanamycin (KMr), and nonpathogenic mutants were selected through the inoculation test on pepper plants. The mutagenesis frequency was about 6$\times$10-8, and 53 out of 2,000 Kmr bacterial colonies tested were nonpathogenic to the pepper cultivar Cheung-Hong. Optimum conditions for the Tn $\Omega$Km mutagenesis of Xcv were Luria Bertani (LB) broth medium for culture of Xcv, yeast extract-dextrose-CaCO3 (YDC) agar medium for selection of Tn $\Omega$Km-mediated mutants, and over 1 to 2 in the ratio of the donor (Escherichia coli S17-1 with the plasmid pJFF350 $\Omega$Km) and the recipient (Xcv) in the culture for the mutagenesis. One of the 4 nonpathogenic mutants (WNP1, WNP3, WNP4 and WNP5), which had been reconfirmed through the inoculation on pepper cv. Dabokgun, showed no differences in the production of exoenzymes such as protease and polygalacturonase and extracellular polysaccharides in vitro and the bacterial growth rate from those of the wild type of Xcv.

• Journal of Applied Biological Chemistry
• /
• v.56 no.2
• /
• pp.109-112
• /
• 2013

Azole fungicides are one of the most wide-spread antifungal compounds in agriculture and pharmaceutical applications. Their major mode of action is the inhibition of ergosterol biosynthesis, giving depletion of ergosterol, precursors and abnormal steroids. However, metabolic consequences of such inhibition, other than steroidal metabolitesare not well established. Comprehensive metabolic profiles of Saccharomyces cerevisiae has been presented in this study. Wild type yeast was treated either with glucose as control or azole fungicide (ketoconazole). Both polar metabolites and lipids were analyzed with gas chromatography-mass spectrometry. Approximately over 180 metabolites were characterized, among which 18 of them were accumulated or depleted by fungicide treatment. Steroid profile gives the most prominent differences, including the accumulation of lanosterol and the depletion of zymosterol and ergosterol. However, the polar metabolite profile was also highly different in pesticide treatment. The concentration of proline and its precursors, glutamate and ornithine were markedly reduced by ketoconazole. Lysine and glycine level was also decreased while the concentrations of serine and homoserine were increased. The overall metabolic profile indicates that azole fungicide treatment induces the depletion of many polar metabolites, which are important in stress response.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.7
• /
• pp.1147-1151
• /
• 2007

Biodegradation of endocrine-disrupting bisphenol A was investigated with several white rot fungi (Irpex lacteus, Trametes versicolor, Ganoderma lucidum, Polyporellus brumalis, Pleurotus eryngii, Schizophyllum commune) isolated in Korea and two transformants of T. versicolor (strains MrP 1 and MrP 13). I. lacteus degraded 99.4% of 50 mg/l bisphenol A in 3 h incubation and 100% in 12 h incubation. which was the highest degradation rate among the fungal strains tested. T. versicolor degraded 98.2% of 50 mg/l bisphenol A in 12 h incubation. Unexpectedly, the transformant of the Mn-repressed peroxidase gene of T. versicolor, strain MrP 1, degraded 76.5% of 50 mg/l bisphenol A in 12 h incubation, which was a lower degradation rate than wild-type T. versicolor. The removal of bisphenol A by I. lacteus occurred mainly by biodegradation rather than adsorption. Optimum carbon sources for biodegradation of bisphenol A by I. lacteus were glucose and starch, and optimum nitrogen sources were yeast extract and tryptone in a minimal salts medium; however, bisphenol A degradation was higher in nutrient-rich YMG medium than that in a minimal salts medium. The initial degradation of endocrine disruptors was accompanied by the activities of manganese peroxidase and laccase in the culture of I. lacteus.

• Journal of Microbiology and Biotechnology
• /
• v.7 no.2
• /
• pp.101-106
• /
• 1997

Strain improvement for the enhanced production of streptokinase and streptodornase in Streptococcus sp. ATCC 12449 was performed. Strain UB111, a hyperproductive mutant which was isolated by use of nitrosoguanidine and selection of colonies with large clear zones on DNase test agar plates supplemented with $1{\%}$ glucose and $0.5{\%}$ ammonium chloride, produced about 3 fold more streptokinase and streptodornase than the wild type when tested in shake flask fermentations. The enhanced production of both streptokinase and streptodornase was achieved by cultivating the mutant in a pH-controlled fermentor containing fermentation medium enriched with yeast extract ($2.1{\%}$). Under these conditions, the mutant produced 7300 units/ml of streptokinase and 800 units/ml of streptodornase.

• Journal of Nutrition and Health
• /
• v.12 no.2
• /
• pp.65-74
• /
• 1979

In order to observe the contents of some essential amino acids in Korean foods, total and free tryptophan, and available lysine in fifty kinds of Korean foods were analyzed by the sfectrophotometry. The results obtained are summarized as follows : 1) The tryptophan contents per 100g of soybean, wet green laver and skim milk were over 250 mg, and the contents Per g nitrogen in ginger, dried persimmon and chestnut were over 150 mg. 2) The free tryptophan contents per 100 g of soybean, small red bean, dried persimmon, ginseng stem were over 100 mg, ana the contents per g nitrogen in dried persimmon was over 150 mg. 3) The available lysine contents Per 100g of soybean, wet green laver and skim milk, fish(auchovy), dried yeast, casein and silkworm pupa were over 1000 mg, and the contents per g nitrogen in potato, perilla(wild sesame), red pepper, sausage and skim milk were over 300mg. 4) The contents of tryptophan and available lysine in soybean, green laver and skim milk were higher than in other samples. 5) In general, the contents of tryptophan and available lysine were abundant in seaweeds.