• Food Science and Preservation
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• v.24 no.3
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• pp.455-463
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• 2017

In this study, the biological activities and physicochemical properties of polysaccharides from Gloiopeltis furcata were investigated. Polysaccharides were isolated by enzymes treatment (celluclast, flavourzyme, papain, termamyl, viscozyme) followed by ethanol precipitation and lyophilization. The yield of polysaccharides by enzymes treatment group were 52.8-66.4%. The major constituents in viscozyme treatment group were total sugar (71.04%), protein (7.22%), uronic acid (23.18 g/100 g), and sulfate (28.27%), respectively. The DPPH radical scavenging activity and ferric reducing antioxidant potential of the viscozyme treatment group at 5 mg/mL were 23.10% and $218.50{\mu}M$, respectively. The protective effects against $H_2O_2$-induced cytotoxicity in L132 cell of viscozyme treatment group at $1{\mu}g/mL$ was 85.64%. The viscozyme treatment group increased the production of nitric oxide (NO) in a dose-dependent manner. The antitumor activity of viscozyme treatment group (at $25{\mu}g/mL$) in A549, HeLa, SNU719 and MCF7 was 69.57%, 52.74%, 61.06% and 68.64%, respectively. All of data showed that the biological activities and chemical characteristics of enzymes treatment group are higher than that of the control group. The polysaccharides isolated from Gloiopeltis furcata investigated herein are useful as functional materials agents.

• Korean Journal of Microbiology
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• v.55 no.2
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• pp.160-163
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• 2019

Senegalimassilia sp. KGMB 04484 was isolated from fecal samples obtained from a healthy Korean. The whole-genome sequence of Senegalimassilia sp. KGMB 04484 was analyzed using the PacBio Sequel platform. The genome comprises a 2,748,041 bp chromosome with a G+C content of 61.18%, 2,300 total genes, 2,139 protein-coding gene, 21 rRNA genes, and 51 tRNA genes. Also, we found that strain KGMB 04484 had some genes for hydrolysis enzyme, fatty acid biosynthesis and metabolism in its genome based on the result of genome analysis. Those genes of KGMB 04484 may be related to regulation of human health and digest.

• Animal Bioscience
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• v.34 no.2
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• pp.172-184
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• 2021

Objective: Vanin1 (VNN1) is a pantetheinase that can catalyze the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies showed that VNN1 is specifically expressed in chicken liver. In this study, we aimed to investigate the roles of peroxisome proliferators activated receptor α (PPARα) and miRNA-181a-5p in regulating VNN1 gene expression in chicken liver. Methods: 5'-RACE was performed to identify the transcription start site of chicken VNN1. JASPAR and TFSEARCH were used to analyze the potential transcription factor binding sites in the promoter region of chicken VNN1 and miRanda was used to search miRNA binding sites in 3' untranslated region (3'UTR) of chicken VNN1. We used a knock-down strategy to manipulate PPARα (or miRNA-181a-5p) expression levels in vitro to further investigate its effect on VNN1 gene transcription. Luciferase reporter assays were used to explore the specific regions of VNN1 targeted by PPARα and miRNA-181a-5p. Results: Sequence analysis of the VNN1 promoter region revealed several transcription factor-binding sites, including hepatocyte nuclear factor 1α (HNF1α), PPARα, and CCAAT/enhancer binding protein α. GW7647 (a specific agonist of PPARα) increased the expression level of VNN1 mRNA in chicken primary hepatocytes, whereas knockdown of PPARα with siRNA increased VNN1 mRNA expression. Moreover, the predicted PPARα-binding site was confirmed to be necessary for PPARα regulation of VNN1 gene expression. In addition, the VNN1 3'UTR contains a sequence that is completely complementary to nucleotides 1 to 7 of miRNA-181a-5p. Overexpression of miR-181a-5p significantly decreased the expression level of VNN1 mRNA. Conclusion: This study demonstrates that PPARα is an important transcriptional activator of VNN1 gene expression and that miRNA-181a-5p acts as a negative regulator of VNN1 expression in chicken hepatocytes.

• Journal of Microbiology and Biotechnology
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• v.32 no.8
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• pp.1064-1071
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• 2022

Arabinogalactans have diverse biological properties and can be used as pharmaceutical agents. Most arabinogalactans are composed of β-(1→3)-galactan, so it is particularly important to identify β-1,3-galactanases that can selectively degrade them. In this study, a novel exo-β-1,3-galactanase, named PoGal3, was screened from Penicillium oxalicum sp. 68, and hetero-expressed in P. pastoris GS115 as a soluble protein. PoGal3 belongs to glycoside hydrolase family 43 (GH43) and has a 1,356-bp gene length that encodes 451 amino acids residues. To study the enzymatic properties and substrate selectivity of PoGal3, β-1,3-galactan (AG-P-I) from larch wood arabinogalactan (LWAG) was prepared and characterized by HPLC and NMR. Using AG-P-I as substrate, purified PoGal3 exhibited an optimal pH of 5.0 and temperature of 40℃. We also discovered that Zn²⁺ had the strongest promoting effect on enzyme activity, increasing it by 28.6%. Substrate specificity suggests that PoGal3 functions as an exo-β-1,3-galactanase, with its greatest catalytic activity observed on AG-P-I. Hydrolytic products of AG-P-I are mainly composed of galactose and β-1,6-galactobiose. In addition, PoGal3 can catalyze hydrolysis of LWAG to produce galacto-oligomers. PoGal3 is the first enzyme identified as an exo-β-1,3-galactanase that can be used in building glycan blocks of crucial glycoconjugates to assess their biological functions.

• Nutrition Research and Practice
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• v.16 no.6
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• pp.685-699
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• 2022

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG) has long been known as a medicinal herb effective in various diseases, including bronchitis and asthma, but is still more widely used for food. Fermentation methods are being applied to increase the pharmacological composition of PG extracts and commercialize them with high added value. This study examines the hydrolyzed and fermented PG extract (HFPGE) fermented with Lactobacillus casei in RAW 264.7 cells, and investigates the effect of amplifying the immune and the probable molecular mechanism. MATERIALS/METHODS: HFPGE's total phenolic, flavonoid, saponin, and platycodin D contents were analyzed by colorimetric analysis or high-performance liquid chromatography. Cell viability was measured by the MTT assay. Phagocytic activity was analyzed by a phagocytosis assay kit, nitric oxide (NO) production by a Griess reagent system, and cytokines by enzyme-linked immunosorbent assay kits. The mRNA expressions of inducible nitric oxide synthase (iNOS) and cytokines were analyzed by reverse transcription-polymerase chain reaction, whereas MAPK and nuclear factor (NF)-κB activation were analyzed by Western blots. RESULTS: Compared to PGE, HFPGE was determined to contain 13.76 times and 6.69 times higher contents of crude saponin and platycodin D, respectively. HFPGE promoted cell proliferation and phagocytosis in RAW 264.7 cells and regulated the NO production and iNOS expression. Treatment with HFPGE also resulted in increased production of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, C-X-C motif chemokine ligand10, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and monocyte chemoattractant protein-1, and the mRNA expressions of these cytokines. HFPGE also resulted in significantly increasing the phosphorylation of NF-κB p65, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. CONCLUSIONS: Taken together, our results imply that fermentation and hydrolysis result in the extraction of more active ingredients of PG. Furthermore, we determined that HFPGE exerts immunostimulatory activity via the MAPK and NF-κB signaling pathways.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.89-93
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• 2011

This study was conducted to investigate the proteolysis and extraction conditions of deer antler for application of food materials. ProteAX (A) was the most effective enzyme for proteolysis of deer antler and the proteolysis condition was 0.5% (w/w) for enzyme concentration and 5 hr for proteolysis time. The effect of mixing enzyme ProteAX (A)+KFEN 2 (C) treatment in $60^{\circ}C$, 5 hr was investigated; soluble solid and protein content were the highest with A 0.5% (w/w) and B 0.5% (w/w) concentration. Result for DAH (deer antler hydrolysate) and DA (deer antler) prepared with extraction in $95^{\circ}C$ atmospheric pressure (AP, 6~18 hr) and extraction under $120^{\circ}C$ pressure condition (UP, 15~60 min) after hydrolysis on preceding established condition descriptions indicated that difference in pH according to enzyme treatment and extraction conditions was not significant. Sugar content of DA was $1.5^{\circ}Brix$, DA-UP (under pressure) and DAH-AP (atmospheric pressure) were $2.2^{\circ}Brix$; the highest sugar content of $2.7^{\circ}Brix$ was observed in DAH-UP for 60 min extraction. Also total free sugar, crude protein and collagen content were the highest in DAH-UP for 60 min recording at 1.97%, 742.7 mg/100 g and 498.8 mg/100 g, respectively. From these results, deer antler hydrolysate prepared with extraction under pressure was the most effective for functional characteristics enhancement. Hereafter, various practical uses of materials with enhanced characteristics of antler is expected.

• Korean Journal of Agricultural Science
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• v.11 no.1
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• pp.120-132
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• 1984

In order to clarify the effect of the fish meat hydrolysate on the growth of lactic acid bacteria(Str. lactis, Str. thermophibus, L. bulgaricus, L. acidophilus, and L. helveticus), the optimum conditions for hydrolyzing the fish meat were examined, and changes of the acid production, viable cell count of lactic acid bacteria and the charge of pH of the culture medium by addition of the fish meat hydrolysate were tested. The results were as follows: 1. When the hydrolysis of back muscle of mackerel was proceeded at $50^{\circ}C$ and at pH 8, for 48 hours adding 6% pancreatin of the protein content in the substrate, the best result was obtained. 2. The composition of the fish meat hydrolysate were 53.6% moisture, 32.4% protein, 1.0% fat, 10.7% carbohydrate, and 3.2% ash. 3. Above 0.1% of the fish meat hydrolysate in the culture medium, the acidity of the culture medium by Sir. lactis and Str. thermophilus were increased remarkably. The acidity of the culture medium by L. acidophilus and L. helveticus were increased in above 0.2% fish meat hydrolysate in the culture medium. but L. bulgaricus was not effected by the fish meat hydrolysate. 4. The pH of the culture medium during incubating Str. laclis and Sir. thermophilus failed obviously by adding the fish meat hydrolysate. But in the cases of L. bulgaricus, L. acidophilus, and L. helveticus, the pH were not changed clearly. 5. The viable cell count in all bacterial strains tested here were elevated by increasing the concentration of the fish meat hydrolysate.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Applied Biological Chemistry
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• v.14 no.1
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• pp.59-97
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• 1971

As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

• Journal of the Korean Wood Science and Technology
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• v.45 no.5
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• pp.599-612
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• 2017

Human hair (HH) is produced as a waste from beauty parlor and barbershop. HH-based adhesives were formulated with NaOH-hydrolyzed HH, $H_2SO_4$-hydrolyzed chicken blood (CB) and PF as a crosslinking agent. Physicochemical properties and retention rate against hot water of the adhesives were measured to investigate the potential of HH as a raw material of wood adhesives. HH was composed of keratin-type protein of 80% and over. Ash of less than 0.1% was contained in HH. Among the amino acids included in HH, glutamic acid showed the highest content, followed by cysteine, serine, arginine and threonine. Solid content of the adhesives ranged from 33.2% to 41.8% depending on hydrolysis conditions of HH and PF type. Viscosity at $25^{\circ}C$ ranged from 300 to $600mPa{\cdot}s$ resulting in a sprayable adhesive. Retention rate against hot water measured to evaluate the water resistance of adhesives was the highest in the cured resin formulated with 5% NaOH-hydrolyzed HH and 5% $H_2SO_4$-hydrolyzed CB. Meanwhile, the molar ratio of formaldehyde to phenol in PF did not have a significant impact on the retention rate of HH-based adhesives. When the retention rates of HH-based adhesives were compared to those of conventional wood adhesive resins used for the production of wood-based panels extensively, HH-based adhesives formulated with 30 wt% PF showed lower retention rate than commercial urea-formaldehyde resin. However, when PF content was increased to 35 wt%, the retention rate greatly increased and approached to that of commercial melamine-urea-formaldehyde resin. Except for the results mentioned above, the analysis of economic feasibility suggests that HH-based adhesives can be used for the production of wood-based panels if HH is hydrolyzed in proper conditions and then the HH-based adhesives are formulated by the HH hydrolyzates with 35 wt% PF.