• Asian-Australasian Journal of Animal Sciences
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• v.23 no.12
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• pp.1632-1638
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• 2010

Two experiments with growing pigs were conducted to investigate the effects of two distinct multienzyme preparations on nutrient digestibility, growth performance and blood profiles. In Exp. 1, a total of 96 pigs ($29.7{\pm}0.69\;kg$) were utilized in a 42-day performance and digestibility trial using four dietary treatments: CON (control diet), ENDO (control+0.10% Endopower), NSPase1 (control+0.10% NSPase) and NSPase2 (control+0.20% NSPase). Endopower was a commercial multienzyme preparation which contained ${\alpha}$-galactosidase, galactomannase, xylanase and ${\beta}$-glucanase. NSPase mainly contained ${\alpha}$-1,6-${\beta}$-galactosidase, ${\beta}$-1,4-mannanase and ${\beta}$-1,4-mannosidase. There were six replication pens per treatment with four pigs per pen. Pigs fed NSPase1 diet had a higher ADG (p<0.05) and G:F (p<0.05) than those fed the control diet. There were no significant differences in growth performance among the multienzyme treatments (p>0.05). Compared with CON, apparent digestibility of DM was increased (p<0.05) by ENDO treatment. N digestibility was improved (p<0.05) in response to multienzyme treatments during the experimental period. Blood urea nitrogen (BUN) was higher (p<0.05) in ENDO treatment than in CON and NSPase1 treatments at the end of the experiment, while the glucose level improved (p<0.05) due to ENDO and NSPase2 treatments. In Exp. 2, four ileal-cannulated, growing barrows ($20.17{\pm}1.31\;kg$) were housed in individual metabolism crates and randomly assigned to 1of 4 treatments (same as Exp. 1) within a $4{\times}4$ Latin square design. Enzyme supplementations improved the majority of apparent ileal amino acid digestibilities (p<0.05). It is concluded that the supplementation of NSPase1 improved growth performance as well as N digestibility and partially improved apparent ileal amino acid digestibility in growing pigs fed a diet based on corn and soybean meal.

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.389-398
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• 2009

Development of expression vectors is important for the basic and applied researches on kimchi LAB (lactic acid bacteria). An expression vector, pSJE6c was constructed by inserting P6C promoter sequence from Lactococcus lactis into pSJE, a shuttle vector for E. coli and Leuconostoc species. To test the efficiency of pSJE6c, aga ($\alpha$-galactosidase) and lacZ ($\beta$-galactosidase) genes were expressed in Lactobacillus brevis 2.14. Compared to the pSJE, expression levels of both genes were increased, indicating P6C promoter was better than indigenous promoters. Enzyme activities of L. brevis cells harboring pSJE6caga (pSJE6c with aga) or pSJE6Z (pSJE6c with lacZ) were 1.5-2 fold higher than those with pSJEaga (pSJE with aga) or pSJEZ (pSJE with lacZ). More RNA transcripts were detected in cells harboring pSJE6c based recombinant plasmid. The results indicated that heterologous gene expressions in kimchi LAB could be improved significantly by use of efficient expression vectors.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.232-237
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• 1998

A bacterial strain producing high levels of an extracellular ${eta}$-mannanase and intracellular ${eta}$-mannosidase and ${alpha}$-galactosidase was isolated from soil. The strain isolated was identified as a strain of Sporolactobacillus sp. and designated as Sporolactobacillus sp. M20l. Synthesis of ${eta}$-mannanase by Sporolactobacillus sp. M20l was induced by sucrose, maltose, or locust bean gum. The highest induction rate was obtained with 2% locust bean gum added to the culture medium as a sole carbon source. On the other hand, induction of ${eta}$-mannosidase was observed only with locust bean gum. The optimal media for the enzyme production were established as follows: for ${eta}$-mannanase; 2% locust bean gum, 0.5% peptone, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 6.0), and for ${eta}$-mannosidase; 2% locust bean gum, 0.5% yeast extract, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 5.0). The optimal culture temperatures for production of ${eta}$-mannanase and ${eta}$-mannosidase were found to be 37$^{\circ}C$ and 3$0^{\circ}C$, respectively. Under the optimal culture conditions, the production of ${eta}$-mannanase and ${eta}$-mannosidase reached the highest levels of 10.6 units/ml and 1.35 units/ml after 30 h and 24 h cultivation, respectively.

• Journal of Oriental Neuropsychiatry
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• v.20 no.1
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• pp.147-164
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• 2009

Objectives : We investigated the effects of Sopungsungi-won(Shu!engshunqiyuan) (SSEx1, SSEx2) to treat the metabolic syndrome by the molecular mechanism of regulation of PPAR and modulation of mitochondrial MCAD, VLCAD mRNA expression. Methods : Mouse NMu2Li liver cells and C2C12 skeletal muscle myogenic progenital cells were transiently transfected with expression plasmids for PPAR(PPAR${\alpha}$, PPAR${\delta}$), a luciferase reporter gene construct containing 3 copies of the PPRE from the rat acyl-CoA oxidase gene and ${\beta}$-galactosidase gene. Cells were treated with several concentrated kinds of SSEx1, SSEx2 at the initial time of culture and analyzed PPAR${\alpha}$, PPAR${\delta}$ reporter gene activity using spectrophotometer (405 nm). Total RNA was extracted from SSEx1, SSEx2 and measured mRNA levels of mitochondrial MCAD, VLCAD. Representative RT-PCR bands are shown. Results : 1. SSEx1 increased the expression of PPAR${\alpha}$ reporter gene activities at 0.1 ${\mu}$g/ml (p${\mu}$g/ml (p<0.05), SSEx2 at 0.1 ${\mu}$g/ml (p${\mu}$g/ml (p<0.05) significantly in NMu2Li liver cell lines. 2. SSEx1 increased the expression of PPAR${\alpha}$ reporter gene activities at 1 ${\mu}$g/ml (p${\mu}$g/ml (p${\alpha}$ reporter gene activities in C2C12 skeletal muscle cells. 4. SSEx1 increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in NMu2Li liver cell lines. 5. SSEx1, SSEx2 both increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in C2C12 skeletal muscle cells. Conclusions : These results show the SSEx1, SSEx2 can be used as therapeutic agent for metabolic syndrome and it's molecular mechanisms of PPAR more contribute to the activation of PPAR${\alpha}$ then PPAR${\delta}$ reporter gene activities and it's total RNA more contribute to the modulation of mitochondrial MCAD then VLCAD mRNA expression.

• Journal of Dairy Science and Biotechnology
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• v.37 no.4
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• pp.223-236
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• 2019

This study aimed to investigate the physiological characteristics and anti-diabetic effects of Lactobacillus plantarum KI69. The α-amylase and α-glucosidase inhibitory activity of L. plantarum KI69 was 91.17±2.23% and 98.71±4.23%, respectively. The propionic, acetic, and butyric acid contents of the MRS broth inoculated with L. plantarum KI69 were 8.78±1.12 ppm, 1.34±0.07% (w/v), and 0.876±0.003 g/kg, respectively. L. plantarum KI69 showed higher sensitivity to penicillin-G, oxacillin, and chloramphenicol among 16 different antibiotics and showed the highest resistance to ampicillin and vancomycin. The strain showed higher β-galactosidase, β-glucosidase, and N-acetyl-β-glucosaminidase activities than other enzymes. Additionally, it did not produce carcinogenic enzymes, such as β-glucuronidase. The survival rate of L. plantarum KI69 in 0.3% bile was 96.42%. Moreover, the strain showed a 91.45% survival rate at pH 2.0. It was resistant to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus with the rates of 15.44%, 50.79%, 58.62%, and 37.85%, respectively. L. plantarum (25.85%) showed higher adhesion ability than the positive control L. rhamnosus GG (20.87%). These results demonstrate that L. plantarum KI69 has a probiotic potential with anti-diabetic effects.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.153-161
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• 2003

This study has evaluated effect of the spermatozoa incubation on the glycosidase activity and fertilizing ability in vitro in the pig. To identify sperm glycosidases specific for sugar residues found in the zona pellucida of pig oocytes, the spermatozoa were treated experimentally and assayed for activities of $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase and N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase). The glycosidases activity were higher in spermatozoa incubated for 2h than without incubation. The $\beta$-GlcNAc'ase activity was at least two-fold higher than other glycosidase regardless of spermatozoa incubation. In the same glycosidases, the activity had a tendency to increase as time of spermatozoa incubation was prolonged, but there were no differences in spermatozoa incubated during the various periods (4~24h). The percentages of spermatozoa that reached acrosome reaction were affected by glycosidases in the medium (P<0.05, for mannosidase), and were higher in spermatozoa with that than without incubation. On the other hand, the spermatozoa motility were decreased with incubation periods, but no effects by different glycosidases on the change of sperm motility during the various periods of incubation. In other experiment, the binding and penetration of pig spermatozoa were tested with oocytes matured in vitro in the presence of various glycosidase. The penetration rates were decreased with incubation of spermatozoa when oocytes were inseminated in medium with different glycosidases. These rates were higher in spermatozoa non-incubated than with incubation for 2h (P<0.05 for GlcNAc'ase; P<0.01 for control group). The sperm-zona binding rate in control group were higherthan in medium with glycosidases. In addition, the highest binding rate were obtained in medium with GlcNAc'ase. In all glycosidases, the sperm-zona binding rate in spermatozoa without incubation were higher than incubation for 2h. The significant differences were obtained in spermatozoa treated with $\alpha$-D-mannosidase (P<0.05). These results suggest that $\beta$-GlcNAc'ase is present mainly in the plasma membrane of pig spermatozoa. It was also shown that the glycosidase activity were increased in all glycosidases in spite of low sperm-zona binding rate and penetration rates by spermatozoa incubation.

• Journal of Life Science
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• v.28 no.11
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• pp.1339-1346
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• 2018

In Korea, edible mushrooms are produced largely on commercial artificial media, so the annual production of spent mushroom substrate (SMS), as a by-product of the mushroom industry, is estimated at over 200 million tons. This SMS is assumed to contain abundant fungal mycelia and pre-fruiting bodies, as well as various nutritive and bioactive compounds that are presently discarded. This study examined the physico-chemical, nutritional, and enzymatic characteristics of uninoculated sterilized medium (USM) and SMS of shiitake mushrooms with the aim of developing a high-value added product from SMS. The contents of crude protein, crude lipid, and ash were higher after the third SMS harvest ($SMS-A-3^{rd}$) than in USM or $SMS-A-1^{st}$. The contents of Ca, Mg, and P in $SMS-A-3^{rd}$ were 2.95, 2.35, and 2.1-fold higher compared than in USM. No As or Cd was detected in USM or SMS. The pH, Brix, and acidity were 4.6, 20.0, and 1.4, respectively in $SMS-A-3^{rd}$, but 5.6, 6.0, and 0.0, respectively, in USM. These results suggest a highly active production of soluble components and organic acids in $SMS-A-3^{rd}$. The distinct color differences noted for USM, $SMS-A-1^{st}$, and $SMS-A-3^{rd}$ could be used as a mycelial growth indicator. Enzyme activity assays using the APIZYM system showed that SMS is a potent source of hydrolysis-related enzymes, especially esterase (C4) and ${\beta}$-glucuronidase. Our results suggested that the SMS of shiitake has a high potential for use in environmental, agricultural, and stock-breeding industries, for example, as active ingredients for sewage treatment, waste-polymer degradation, and feed additives.

• Journal of Genetic Medicine
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• v.4 no.1
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• pp.45-52
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• 2007

Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

• Korean Journal of Ecology and Environment
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• v.36 no.2 s.103
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• pp.108-116
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• 2003

To investigate bacteria with algal Iytic activities against Anabaena cylindrica when water blooming occurs and to study enzyme profiles of alga-Iytic bacteria, various bacterial strains were isolated from surface waters and sediments in eutrophic lakes or reservoirs in Korea. Abacterial strain AK-07 was characterized and identified as Acinetobacter johnsonii based on its16S rDNA base sequence. When AK-07 was co-cultivated with A. cylindrica, bacterial cells propagated to $8\;{\times}\;10^8$ cfu $ml^{-1}$ and Iyses algal cells. However, culture filtrates of AK-07 did not exhibit algal Iytic activities. That suggesting the enzymes on the surfaces of the bacterium might be effective algal Iytic agents to cause Iyses of cells. Acinetobacter johnsonii AK-07 exhibited high degradation activities against A. cylindrica, and formed alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, glycosidases for example ${\beta}$-galatosidase, ${\beta}$-glucosidase, ${\beta}$-glucosaminidase, and ${\beta}$-xylosidase, which hydrolyzed ${\beta}$-0-glycosidic bonds, were found in cell-free extracts of A. johnsonii AK-07. Other glycosidase such as ${\alpha}$-galctosidases, ${\alpha}$-N-Ac-galctosidases, ${\alpha}$-mannosidases, and ${\alpha}$- L-fuco-sidases, which cleavage ${\alpha}$-0-glycosidic bondsare not detected. In the results, enzyme systemsof A. johnsonii AK-07 were very complex to do-grade cell walls of cyanobacteria. The polysaccharides or peptidoglycans of A. cylindrica maybe hydrolyzed and metabolized to a range of easily utilizable monosaccharides or other low molecular weight organic substances by strain AK-07 of A. johnsonii.

• Food Science of Animal Resources
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• v.38 no.3
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• pp.554-569
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• 2018

This study aimed to investigate the physiological characteristics and anti-obesity effects of Lactobacillus plantarum K10. The ${\alpha}-amylase$ inhibitory activity, ${\alpha}-glucosidase$ inhibitory activity, and lipase inhibitory activity of L. plantarum K10 was $94.66{\pm}4.34%$, $99.78{\pm}0.12%$, and $87.40{\pm}1.41%$, respectively. Moreover, the strain inhibited the adipocyte differentiation of 3T3-L1 cells ($32.61{\pm}8.32%$) at a concentration of $100{\mu}g/mL$. In order to determine its potential for use as a probiotic, we investigated the physiological characteristics of L. plantarum K10. L. plantarum K10 was resistant to gentamycin, kanamycin, streptomycin, ampicillin, ciprofloxacin, tetracycline, vancomycin, and chloramphenicol. It also showed higher Leucine arylamidase, Valine arylamidase, and ${\beta}-galactosidase$ activities. Moreover, it was comparatively tolerant to bile juice and acid, exhibiting resistance to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus with rates of 90.71%, 11.86%, 14.19%, and 23.08%, respectively. The strain did not produce biogenic amines and showed higher adhesion to HT-29 cells compared to L. rhamnosus GG. As a result of the animal study, L. plantarum K10 showed significantly lower body weight compared to the high-fat diet group. The administration of L. plantarum K10 resulted in a reduction of subcutaneous fat mass and mesenteric fat mass compared to the high-fat diet (HFD) group. L. plantarum K10 also showed improvement in gut permeability compared to the HFD positive control group. These results demonstrate that L. plantarum K10 has potential as a probiotic with anti-obesity effects.