• Asian-Australasian Journal of Animal Sciences
• /
• v.27 no.8
• /
• pp.1131-1140
• /
• 2014

This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight $7.14{\pm}0.63$ kg) weaned at $26{\pm}2$ days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs per pen) for a 28-day trial. Dietary treatments were basal diet without any antimicrobial (negative control; NC), basal diet supplemented with 120 mg/kg feed of neomycin sulfate (positive control; PC) and basal diet supplemented with $2{\times}10^9$ (L), $4{\times}10^9$ (M) and $20{\times}10^9$ (H) CFU/kg feed of B. subtilis KN-42. During the overall period, average daily gain and feed efficiency of piglets were higher in groups PC, M, and H than those in group NC (p<0.05), and all probiotics and antibiotics groups had a lower diarrhea index than group NC (p<0.05). The 16S rDNA gene-based methods were used to analyze faecal bacterial flora on day 28 of experiment. The result of denaturing gradient gel electrophoresis analysis showed that supplementation of B. subtilis KN-42 to the diet changed the bacterial communities, with a higher bacterial diversity and band number in group M than in the other four groups. Real-time polymerase chain reaction analysis showed that the relative number of Lactobacillus were higher in groups PC and H than in group NC (p<0.05), and the supplemented B. subtilis KN-42 to the diet also reduced the relative number of E. coli (p<0.05). These results suggest that dietary addition of B. subtilis KN-42 can improve the growth performance and gastrointestinal health of piglets.

• Journal of Life Science
• /
• v.22 no.4
• /
• pp.559-564
• /
• 2012

In order to investigate the effect of phytochromes on the regulation of ethylene biosynthesis, we measured the ethylene production and the activities of enzymes involved in ethylene biosynthesis using phytochrome mutants such as $phyA$, $phyB$, and $phyAB$ of Arabidopsis. The ethylene production was decreased in mutants grown in white light. In particular, double mutants showed a 37% decrease compared to the wild type in ethylene production. When Arabidopsis roots were grown in the dark, mutants did not show a decrease in ethylene production; however, production was significantly decreased in the double mutant grown in red light. Only $phyB$ did not show the decrease in the ethylene production in far-red light. Unlike the ACO activities, the ACS activities of mutants showed the same pattern as the ethylene production under several light conditions. The results of ACS activities confirmed the expression of the ACS gene by RT-PCR analysis. The decrease of ethylene production in mutants was due to the lower activity of ACC synthase, which converts the S-adenosyl-L-methionine (AdoMet) to 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene. These results suggested that both phytochrome A and B play an important role in the regulation of ethylene biosynthesis in Arabidopsis roots in the conversion step of AdoMet to ACC, which is regulated by ACS.

• Korean Journal of Food Science and Technology
• /
• v.44 no.5
• /
• pp.600-606
• /
• 2012

A Bacillus sp. that produces fibrinolytic enzyme was isolated from Cheonggukjang, a traditional Korean soybean-fermented food. According to 16S rRNA gene base sequencing, the bacillus was identified as a variety of Bacillus subtilis, and named Bacillus subtilis IDCC 9204. Fibrinolytic enzyme NK-IL9204 was stable up to $60^{\circ}C$ and within pH range of 5-10. Purified NK-IL9204 was detected through fibrin zymography. The molecular weight and isoelectric point of the enzyme were estimated to be 27.7 kDa and 6.7 by SDS-PAGE and 2D electrophoresis, respectively. Its amino acid sequence was similar to that of nattokinase (identities 99.5%) and different from that of nattokinase BPN (identities 86.4%). The plasma fibrinolytic activity of NK-IL9204 was measured by euglobulin clot lysis times (ECLT). The NK-IL9204 was orally administered to SD rats for 3 weeks (1,000 FU/rat/day). The ECLT was significantly shortened by supplementation of NK-IL9204.

• KSBB Journal
• /
• v.29 no.5
• /
• pp.316-322
• /
• 2014

Xylanase have been used to convert the polymetric xylan into fermentable sugars from the production of ethanol and xylitol from plant biomass. The aim of this study was to isolate and identify xylanolytic bacterium from herbivore feces and was to used the xylanase for enzymatic hydrolysis of biomass. Xylanolytic strains were isolated from 59 different feces of herbivores from Seoul Grand Park located in Gwacheon Gyeonggi-do. The xylanolytic strains were selected by congo red staining and DNS method. Total 67 strains isolated from the herbivores feces were tested for xylanase activity. Among the strains, H10-1, which has the highest xylanase activity, was isolated from feces of Ceratotherium simum. The H10-1 strain was identified as Bacillus pumilus based on its morphological/biochemical characteristics and partial 16S rDNA gene sequences. Culture conditions of B. pumilus H10-1 such as initial medium pH, incubation temperature and incubation time were optimized for maximum xylanase production. And also xylanase produced by B. pumilus H10-1 was applied for the saccharification of Miscanthus sacchariflorus cv. 'Geodae 1', which was pretreated with 1.5M NaOH. The optimized culture conditions of B. pumilus H10-1 were pH 9, $30^{\circ}C$ incubation temperature, and 7 day incubation time, respectively. This xylanase activity under the optimized conditions was $20.4{\pm}3.3IU$. The crude xylanase produced by B. pumilus H10-1 was used for the saccharification of xylan derived from pretreated 'Geodae 1'. The saccharification conditions were $50^{\circ}C$, 200 rpm, and 5 days. Saccharification efficiency of pretreated 'Geodae 1' by B. pumilus H10-1 was 8.2%.

• The Korean Journal of Mycology
• /
• v.43 no.3
• /
• pp.200-205
• /
• 2015

Perilla frutescens var. japonica Hara was collected from farmland in Seongju-gun. Fifteen endophytic fungal strains with different colony morphologies were isolated from roots of P. frutescens. Waito-c rice seedlings were treated with the concentrated culture filtrates (CF) of endophytic fungi for observation of their plant growth-promoting activities. In the results, the CF of Y2H001 fungal strain promoted the growth of the waito-c rice seedlings. The phylogenetic tree of Y2H001 strain was analyzed by the combined sequences of the partial internal transcribed spacer region (ITS) and partial betatubulin gene. Molecular and morphological studies identified the Y2H001 strain as belonging to Aspergillus flavus. In gas chromatography mass spectrometry (GC/MS) analysis of the CF of Y2H001 strain, gibberellic acid (GA) was detected and quantified. Therefore, we describe Y2H001 strain as a new $GA_3$-producing A. flavus based on morphological, molecular characteristics and analysis of secondary metabolite.

• Microbiology and Biotechnology Letters
• /
• v.48 no.1
• /
• pp.57-63
• /
• 2020

Over the past 20 years, global warming has transformed the marine ecosystem of the Jeju Island into a subtropical zone making it conducive to the production of tropical fishes. Recently, the balloon fish (Diodon holoanthus) has been found off the coast of the Jeju Island. In this study, we analyzed the diversity of its intestinal microorganisms as a representative for the surrounding environment. In addition, the isolates were evaluated for their antibacterial activity. A total of 161 strains of various species were identified and isolated using 16S ribosomal RNA gene sequence analysis. They were separated into three groups, of which Phylum Proteobacteria was found to be the most dominant with 91% sequence similarity. This includes the class γ-proteobacteria that is made up of twelve genera and twenty-four hundred species. The second group comprised strains of the genus Vibrio, made up of 35% Photobacteria, 32% Shewanella, and 6% Psychrobacter. It was also determined that 4% of the isolates were Acinetobacter, 3% were Enterovibrio, while Moraxella_g2 accounted for 1% of the total isolates. Class α-proteobactera includes five genera and five species; Brevundimonas, Allorhizobium, Pseudoceanicola and Erythrobcter, each accounting for 1% of the total isolates. The Firmicute strains belonged to six genera and ten species. 5% of the strains were Terribacillus, while Paenibacillus, Salinicoccus, Staphylococcus and Streptococcus accounted for 1% each of the total isolates. Actinobacteria accounted for the final phylum with strains belonging to three genera and ten species with Janibacter, Micrococcus and Isoptericola each accounting for 1% of the total isolates.

• Journal of Plant Biotechnology
• /
• v.35 no.4
• /
• pp.317-327
• /
• 2008

Phytocystatins, which are inhibitors of plant cysteine peptidases, are involved in the regulation of protein turnover and in the defense against insect pests and pathogens. Extensive searches in the Brassica rapa genome allowed the prediction of at least eight different phytocystatin genes on seven chromosomes in the B. rapa genome. Structure comparisons based on alignments of the all BrCYS ($\underline{B}$. $\underline{r}apa$ $phyto{\underline{cys}}tatin$) proteins using the CLUSTALW program revealed conservation of the three consensus motifs known to interact with the active site of cysteine peptidases. According to the phylogenetic analysis based on the deduced amino acid sequences, the eight BrCYS proteins were divided into several clusters related to the orthologous phytocystatin. The predicted three-dimensional structure models of the eight BrCYS proteins demonstrate that all of these proteins are similar to the reported crystal structure of oryzacystatin-I (OC-I). Digital northern and RT-PCR analyses indicated that the eight BrCYS genes exhibit different expression patterns in B. rapa tissues and respond differently to abiotic stimuli. The differences in gene structure and expression between the eight BrCYS genes suggest that these proteins may play diverse physiological roles in B. rapa and may interact with cysteine peptidases through different mechanisms.

• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.5
• /
• pp.788-801
• /
• 2020

Objective: This study was conducted to evaluate the effects of diets containing different wet rice distillers' by-product (RDP) levels on growth performance, nutrient digestibility, blood profiles and gut microbiome of weaned piglets. Methods: A total of 48 weaned castrated male crossbred pigs, initial body weight 7.54±0.97 kg, and age about 4 wks, were used in this experiment. The piglets were randomly allocated into three iso-nitrogenous diet groups that were fed either a control diet, a diet with 15% RDP, or a diet with 30% RDP for a total of 35 days. Chromium oxide was used for apparent digestibility measurements. On d 14 and d 35, half of the piglets were randomly selected for hemato-biochemical and gut microbiota evaluations. Results: Increasing inclusion levels of RDP tended to linearly increase (p≤0.07) average daily gain on d 14 and d 35, and decreased (p = 0.08) feed conversion ratio on d 35. Empty stomach weight increased (p = 0.03) on d 35 while digestibility of diet components decreased. Serum globulin concentration decreased on d 14 (p = 0.003) and red blood cell count tended to decrease (p = 0.06) on d 35, parallel to increase RDP levels. Gene amplicon profiling of 16S rRNA revealed that the colonic microbiota composition of weaned pigs changed by inclusion of RDP over the period. On d 14, decreased proportions of Lachnospiraceae_ge, Ruminococcaceae_ge, Ruminococcaceae_UCG-005, and Bacteroidales_ge, and increased proportions of Prevotellaceae_ge, Prevotella_2, and Prevotella_9 were found with inclusion of RDP, whereas opposite effect was found on d 35. Additionally, the proportion of Lachnospiraceae_ge, Ruminococcaceae_ge, Ruminococcaceae_UCG-005, and Bacteroidales_ge in RDP diets decreased over periods in control diet but increased largely in diet with 30% RDP. Conclusion: These results indicate that RDP in a favorable way modulate gastrointestinal microbiota composition and improve piglet performance despite a negative impact on digestibility of lipids and gross energy.

• Molecules and Cells
• /
• v.25 no.1
• /
• pp.55-63
• /
• 2008

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.

• Journal of Animal Science and Technology
• /
• v.46 no.3
• /
• pp.495-502
• /
• 2004

Intimin, the product of eae gene in EHEC O157:H7, is required for intimate adherence. In this study, the C-terminaI region(281 amino acids) of the EHEC OI57:H7 intimin were expressed as a protein fusion with (His)$_6$ which was used to raise antiserum in rabbits. The antiserum reacted in western blot with a 94kDa outer membrane protein of EHEC O157:H7. It was observed that the antibody titers both in egg yolk and serum appeared in 2${\sim}$4 weeks after immunization with fusion protein. At the time of 8 weeks, the titre of egg yolk was found to be higher than that of sera. According to the results of neutralization test, chicken egg-yolk antibody(lgY) against the recombinant intimin strongly reacted to EHEC O157:H7. We conclude that a truncated recombinant intimin could be used as an immunogen to elicit antibody(lgY) against O157:H7.