• Journal of Dairy Science and Biotechnology
• /
• v.35 no.3
• /
• pp.208-214
• /
• 2017

The aims of this study were to manufacture a hydrolyzed goat milk protein (HGMP)/chitosan ologisaccharide (CSO) nano-delivery system (NDS) and to investigate the effects of production variables, such as sodium tripolyphosphate (TPP), HGMP, and CSO concentration levels, on the formation and physicochemical properties of the NDS. An HGMP/CSO NDS was produced using the ionic gelation method at pH 5.5. Transmission electron microscopy and a particle size analyzer were used to determine the morphological and physicochemical properties of NDSs, respectively. The size of the HGMP/CSO NDS decreased from 225 to 138 nm as HGMP and CSO concentration levels decreased. The NDS had a positive surface charge, with a zeta-potential value of +23 mV. The encapsulation efficiency (EE) of docosahexaenoic acid was enhanced as the HGMP concentration level increased. Additionally, increasing the concentration level of CSO resulted in an increase in the EE of resveratrol. The HGMP/CSO NDS exhibited good physical stability during freeze-drying. Thus, our findings showed that the HGMP/CSO NDS was successfully manufactured and that HGMP and CSO concentration levels were key factors affecting the physicochemical properties of the NDS.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.4
• /
• pp.556-562
• /
• 2007

A total of 126 crossbred weanling pigs (average body weight of $6.3{\pm}0.3$ kg) were used to investigate the effect of chito-oligosaccharide (COS) on growth performance, nutrient digestibility, pH of gastro-intestinal tract (GI), intestinal and fecal microflora of young piglets. Pigs were allocated to three dietary treatments based on body weight and gender in a single factorial arrangement. Treatments were control (No COS), T1 (0.2% COS during starter (6-13 kg) and 0.1% COS during grower (13-30 kg) phases, and T2 (0.4% COS during starter (6-13 kg) and 0.3% COS during grower (13-30 kg) phases, respectively. Each treatment had 3 replicates and 14 pigs were raised in each pen. COS is a low molecular weight water-soluble chitosan that can be obtained from chitin of the crab shell after deacetylation with concentrated sodium hydroxide at high temperature and then further decomposition by chitosanase enzyme in the presence of ascorbic acid. For the starter and grower periods, there were no significant differences (p>0.05) in average daily gain (ADG) and feed to gain ratio among treatments. However, during the overall period (6-30 kg), T2 showed better (p<0.05) feed to gain ratio than other treatments. A digestibility study was conducted at the end of grower phase which showed improvement (p<0.05) in DM and crude fat digestibility in T2 over the control. At 25 kg body weight, 6 pigs per treatment (2 per replicate) were sacrificed to determine the effect of diets on pH and microbial count at different sections of the GI tract. The pH of the cecal contents in pigs fed 0.1% COS was higher (p<0.05) than in the other treatments. Total anaerobic bacterial number increased from cecum to rectum in all treatments. The weekly total bacterial counts showed higher (p<0.05) in feces of pigs fed COS than that of untreated pigs at the $8^{th}$ week. The number of fecal E. coli in untreated pigs at $4^{th}$ wk was 7.35 log CFU/g compared to 6.71 and 6.54 log CFU/g in 0.1 and 0.3% COS-treated pigs, respectively. Similarly, at $8^{th}$ wk, fecal clostridium spp. were lower in pigs fed 0.3% COS (5.43 log CFU/g) than in untreated pigs (6.26 log CFU/g). In conclusion, these results indicated that chito-oligosaccharide could improve feed efficiency in young pigs and inhibited the growth of harmful bacteria.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.11
• /
• pp.1651-1658
• /
• 2008

This study was conducted to determine the effects of dietary supplementation of either 100 mg/kg chito-oligosaccharide (COS) or chlortetracycline (CTC) with corn-soybean-fish meal on immunity in broiler chickens. A total of 147 one-day old male broiler chicks were randomly allocated to 3 treatments with 7 replicate pens per treatment and 7 birds per pen. The experimental diets consisted of a control diet based on corn, soybean and fish meal without COS and any antibiotic supplement and similar diets supplemented with either CTC (80 mg/kg from d 1 to 21 and 50 mg/kg from d 22 to 42) or COS (100 mg/kg from d 1 to 42). During the entire experimental period, all birds had ad libitum access to diets and water. The main immune organ indices, T-lymphocyte proliferation, serum cytokine concentrations, serum NO level and serum iNOS activity were measured on d 21 and d 42. On d 21, broilers fed 100 mg/kg COS had improved (p<0.01) indices of spleen, thymus, and bursa of Fabricius compared with the control and CTC birds. Birds receiving 100 mg/kg COS had higher (p<0.05) serum concentrations of $IL-1{\beta}$, IL-6, IgM, NO and iNOS than birds on the control treatment. Serum $Ca^{2+}$ level of birds fed 100 mg/kg COS tended to be higher (p = 0.049) than in birds fed CTC. On d 42, the birds fed 100 mg/kg COS had higher (p<0.05) concentrations of TNF-${\alpha}$ and IgM in serum than birds in both the CTC and control treatments. Birds fed 100 mg/kg COS had a higher concentration of IFN-$\gamma$ than the control group. In conclusion, dietary supplementation of COS appeared to improve the immunity of broilers by promoting the weight of the main immune organs, increasing IgM secretion, stimulating microphages to release $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 and IFN-$\gamma$, and activating iNOS to induce NO.

• Food Science and Preservation
• /
• v.6 no.4
• /
• pp.514-520
• /
• 1999

Changes in pH, titratable acidity, CO$_2$production, reducing sugar, and lactic acid bacteria of fresh kimchi juice supplemented with combinations of 3 lactic acid bacteria and germanium(Ge)-132 or fructooligosaccharide(FO) during fermentation at 30$^{\circ}C$ were investigated to assess the potential for extending the shelf life and enhancing the functional properties in kimchi. In kimchi juice containing Ge-132, sample(I) (inoculated with a mixture of bacteriocin-producing SNF-13 strain and E. faecium) exhibited that the amounts of organic acid and evolved CO$_2$gas were lower than those of the other starter samples(II-IV). The growth of lactic acid bacteria naturally present in kimchi juice, particularly Lb. plantarum and Leu. mesentroides, may be inhibited due to competition of the isolated SNF-13 strain and E. faecium by Ge-132. During fermentation of kimchi juice containing FO sugar, the contents of organic acid and evolved CO$_2$gas On juice broth with 4 starters were predominantly higher than those of control and Ge-132 groups, and then the growth of lactic acid bacteria originated from kimchi ingredients was thought to be markedly accelerated. Our results indicated that functional properties like the extension of shelf life and increase of biological activity in kimchi were enhanced by adding Ge-132 and bacterium-producing lactic acid bacterium, which were resistant to organic acid and stimulated by Ge-132.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.1
• /
• pp.73-79
• /
• 2004

The objective of the present study was to determine whether ${\beta}$1-4 galacto-oligosaccharides (GOS) and Candida kefyr combined with nitrate as manipulators could suppress rumen methanogenesis without nitrate poisoning in sheep. Four rumen fistulated wethers were allocated to a $4{\times}4$ Latin square design. Nitrate (1.3 g $NaNO_3$ $Kg^{-0.75}$body weight) with and without GOS and Candida kefyr were administered into the rumen through fistula as a single dose 30 min after the morning meal. GOS and Candida kefyr were supplemented by sprinkling onto the feed and through rumen fistula, respectively. The four treatments consisted of saline, nitrate, nitrate plus GOS and nitrate plus GOS plus Candida kefyr. Physiological saline was used as the control treatment. Compared to saline treatment, the administration of nitrate alone resulted in a very marked decrease in rumen methanogenesis and an increase in rumen and plasma nitrite production and blood methaemoglobin formation consequently causing a decline in oxygen consumption, carbon dioxide production and metabolic rate. When compared to nitrate alone, the simultaneous administration of nitrate with GOS decreased nitrite accumulation in rumen and plasma and nitrate-induced methaemoglobin, while retaining low methane production. However, GOS could not fully restore metabolic parameters reduced by nitrate. When compared to the simultaneous administration of nitrate with GOS, the simultaneous administration of nitrate with GOS plus Candida kefyr lowered rumen methanogenesis to a negligible level, but did not decrease rumen and plasma nitrite accumulation as well as blood methaemoglobin formation. Thus, these results suggest that combination of nitrate with GOS may be a potent manipulator to suppress rumen methanogenesis with abating the hazards of nitratenitrite toxicity in ruminants.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.10
• /
• pp.1153-1160
• /
• 2009

Lactosucrose ($4^G-\beta$-D-galactosylsucrose) is an oligosaccharide consisting of galactose, glucose, and fructose. In this study, we prepared lactosucrose from lactose and sucrose using a levansucrase derived from Zymomonas mobilis. Optimum conditions for lactosucrose formation were $23^{\circ}C$, pH 7.0, 18.0% (w/v) lactose monohydrate, and 18% (w/v) sucrose as substrates, and 1 unit of enzyme/ml of reaction mixture. Under these conditions, the lactosucrose conversion efficiency was 28.5%. The product was purified and confirmed to be O-$\beta$-D-galactopyranosyl-($1{\rightarrow}4$)-O-$\beta$)-D-glucopyranosyl-($1{\rightarrow}2$)-$\beta$-D-fructofuranoside, or lactosucrose. A mixed-enzyme system containing a levansucrase and a glucose oxidase was applied in order to increase the efficiency of lactose and sucrose conversion to lactosucrose, which rose to 43.2% as a result.

• Reproductive and Developmental Biology
• /
• v.33 no.2
• /
• pp.77-83
• /
• 2009

Erythropoietin (EPO), a glycoprotein hormone produced from primarily cells of the peritubular capillary endothelium of the kidney, is responsible for the regulation of red blood cell production. We have been investigating the roles of glycosylation site added in the biosynthesis and function of recombinant protein. We constructed three EPO mutants ($\Delta$69, $\Delta$105 and $\Delta$69,105), containing an additional oligosaccharide chains. EPOWT and EPO$\Delta$69 were effectively expressed in transient and stably transfected CHO-K1 cell lines. But, it wasn't detected any protein in the culture medium of EPO$\Delta$105 and EPO$\Delta$69,105 mutants. The growth and differentiation of EPO-dependent human leukemic cell line (F36E) were used to measure the cytokine dependency and in vitro bioactivity of rec-hEPO. MTT assay values were increased by survival of F36E cells at 24h. To analysis biological activity in vivo, two groups of ICR-mice (7 weeks old) were injected subcutaneously with 10 IU per mice of rec-hEPO molecules on days 0 and 2. Red blood cell and hematocrit values were measured on 6 days after the first injection. The hematocrit values were remarkably increased in all treatment groups. The pharmacokinetic analysis was also affected in the mice injected with rec-hEPO molecules 2.5 IU by tail intravenous. Protein samples were detected by Western blotting. An EPO$\Delta$69 protein migrated as a broad band with an average apparent molecular and detected slightly high band. Enzymatic N-deglycosylation resulted in narrow band and was the same molecular size. The biological activity of EPO$\Delta$69 was enhanced to compare with wt-hEPO. The half-life was longer than wt-hEPO. The results suggest that hyperglycosyalted recombinant human erythropoietin (EPO$\Delta$69) may have important biological and therapeutic good points.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.9
• /
• pp.1237-1244
• /
• 2012

Agarase genes of non-marine agarolytic bacterium Cellvibrio sp. were cloned into Escherichia coli and one of the genes obtained using HindIII was sequenced. From nucleotide and putative amino acid sequences (713 aa, molecular mass; 78,771 Da) of the gene, designated as agarase AgaA, the gene was found to have closest homology to the Saccharophagus degradans (formerly, Microbulbifer degradans) 2-40 aga86 gene, belonging to glycoside hydrolase family 86 (GH86). The putative protein appears to be a non-secreted protein because of the absence of a signal sequence. The recombinant protein was purified with anion exchange and gel filtration columns after ammonium sulfate precipitation and the molecular mass (79 kDa) determined by SDS-PAGE and subsequent enzymography agreed with the estimated value, suggesting that the enzyme is monomeric. The optimal pH and temperature for enzymatic hydrolysis of agarose were 6.5 and $42.5^{\circ}C$, and the enzyme was stable under $40^{\circ}C$. LC-MS and NMR analyses revealed production of a neoagarobiose and a neoagarotetraose with a small amount of a neoagarohexaose during hydrolysis of agarose, indicating that the enzyme is a ${\beta}$-agarase.

• KSBB Journal
• /
• v.14 no.6
• /
• pp.707-712
• /
• 1999

We have produced new-structured oligosaccharides using mixed-enzyme reactor of dextransucrase from Leuconostoc mesenterides B-512FMCM and ${\alpha}$-amylase. When the concentrations of sucrose and starch were 10%(w/v) and 5%(w/v), respectively, the maximum yield of oligosaccharides with both dextransucrase(100U) and ${\alpha}$-amylase(1000U) was 66.4%. The activity of dextransucrase in mixed-enzyme reactor was increased about 2.5 times by acceptor reaction with starch hydrolyzates. As the activities of dextransucrase:${\alpha}$-amylase were increased from 20U:200U to 500U:5000U, the amount of polymer was increased and the yield of oligosaccharides was decreased. By the addition of sucrose into mixed-enzyme reactor following the prehydrolysis of starch with ${\alpha}$-amylase, the yield was increased up to 12% compared with that of mixed-enzyme reactor without the addition of starch hydrolyzate. New structured-oligosaccharides showed heat resistance up to 140$^{\circ}C$ and was stable in acidic condition at pH 3~6.

• BMB Reports
• /
• v.31 no.1
• /
• pp.53-57
• /
• 1998

The CMCax gene from Acetobacter xylinum ATCC 23769 was cloned and expressed in E. coli. With this gene, three gene products - mature CMCax, CMCax containing signal peptide(pre-CMCax), and a glutathione-S-transferase(GST)-CMCax fusion enzyme - were expressed. CMCax and pre-CMCax are aggregated to multimeric forms which showed high CMC hydrolysis activity, whereas GST-CMCax was less aggregated and showed lower activity, indicating that oligomerization of CMCax controbutes to the cellulose hydrolysis activity to achieve greater efficiency. The enzyme was identified to be an $\beta$-1,4-endoglucanase, which catalyzes the cleavage of internal $\beta$-1,4-glycosidic bonds of cellulose. The reaction products, cellobiose and cellotriose, from cellopentaose as a substrate, were identified by HPLC. Substrate specificity of cellotetraose by this enzyme was poor, and the reaction products consisted of glucose, cellobiose, and cellotriose in a very low yield. Theses results suggested that cellopentaose might be the oligosaccharide substrate consisting of the lowest number of glucose. The optimum pH of CMCax and pre CMCax was about 4.5, whereas that of GST-CMCas was rather broad at pH 4.5-8. The physiological significance of cellulose-hydrolyzing enzyme, CMCax, having such low $\beta$-1,4-endoglucanase activity and low optimum pH in cellulose-producing A. xylinum is not clearly known yet, but it seems to be closely related to the production of cellulose.