Corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. Recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. In this study, we conducted the optimization of medium components for improved succinate production from C. glutamicum under an oxygen deprivation condition by Plackett-Burman design and applied a response surface methodology. A Plackett-Burman design for ten factors such as glucose, ammonium sulfate, magnesium sulfate, potassium phosphate ($K_2HPO_4$ and $KH_2PO_4$), iron sulfate, manganese sulfate, biotin, thiamine, and sodium bicarbonate was applied to evaluate the effects on succinate production. Glucose, ammonium sulfate, magnesium sulfate, and dipotassium phosphate were found to have significant influence on succinate production, and the optimal concentrations of these four factors were sequentially investigated by the response surface methodology using a Box-Behnken design. The optimal medium components obtained for achieving maximum concentration of succinic acid were as follows: glucose 10 g/l, magnesium sulfate 0.5 g/l, dipotassium phosphate ($K_2HPO_4$) 0.75 g/l, potassium dihydrogen phosphate ($KH_2PO_4$) 0.5 g/l, iron sulfate 6 mg/l, manganese sulfate 4.2 mg/l, biotin 0.2 mg/l, thiamine 0.2 mg/l, and sodium bicarbonate 100 mM. The parameters that differed from a normal BT medium were glucose changed from 40 g/l to 10 g/l, dipotassium phosphate ($K_2HPO_4$) 0.5 g/l changed to 0.75 g/l, and ammonium sulfate ($(NH_4)_2SO_4$) 7 g/l changed to 0 g/l. Under these conditions, the final succinic acid concentration was 16.3 mM, which is about 1.46 fold higher than the original medium (11.1 mM) at 24 h. This work showed the improvement of succinate production by a simple change of media components deduced from sequential optimization.
The protease produced by a newly isolated Aspergillus wentti from Korean traditional Meju was purified and characterized. The optimal medium composition and culture conditions for maximum protease production were ; bran :1% glucose solution =1 : 1, pH 9.0, $30^{\circ}C$, and 4 days of fermentation. Protease was purified by QAE-Sephadex, SP-Sephadex ion exchange chromatography and Sephadex G-100 chromatography. The specific activity and the purification fold of the purified enzyme were 213 unit/mg protein and 27.3, respectively. The molecular weight of purified protease was found to be 32 kDa by SDS-PAGE. Km and Vmax value's for hammastein milk casein were $3.049{\times}10^{-4}\;M\;and\;151.1\;{\mu}g/min$, respectively. Kinetic parameters showed that the enzyme has higher affinity to casein than isolated soybean protein, hemoglobin and bovine serum albumin. Optimal pH and temperature for reaction of the purified enzyme were 9.0 and $50^{\circ}C$, respectively. The enzyme was stable at pH 4.0-11.0, below $40^{\circ}C$, and the activity was not stimulated by metal ions. 1mM phenylmethylsulfonyl fluoride inhibited the enzyme activity by 98.5%. It means that the enzyme is one of serine protease.
A vector harboring double cassettes; a heterologous gene expression cassette of pHCE-InaN-antigen and a ghost formation cassette of pAPR-cI-E lysis 37 SDM was constructed and introduced to E. coli DH5a. For the production of a bacterial ghost vaccine, bacterial ghosts from E. coli / Streptococcus iniae with four different types of antigens - enolase, GAPDH, sagA and piaA - were produced by the optimization of fermentation parameters such as a glucose concentration of 1 g/l, agitation of 300 rpm and aeration of 1 vvm. Efficiency of ghost bacteria formation was evaluated with cultures of OD$_{600}$=1.0, 2.0 and 3.0. The efficiency of the ghost bacteria formation was 99.54, 99.67, 99.99 and 99.99% with inductions at OD$_{600}$=3.0, 1.0, 2.0 and 1.0 for E. coli/S. iniae antigens enolase, piaA, GAPDH and sagA, respectively. Ghost bacteria as a vaccine was harvested by centrifugation. The antigen protein expressions were analyzed by SDS-PAGE and western blot analysis, and the molecular weights of the enolase, piaA, GAPDH and sagA were 78, 26, 67 and 26 kDa, respectively. The molecular weights of the expressed antigens were consistent with theoretical sizes obtained from the amino acid sequences.
This study was carried out to estimate the effects of preservatives (alcohol, adipic acid) and heat treatment $(40^{\circ}C,\;60^{\circ}C)$ for the prolongation of shelf-life on low-salt Kimchi. Low-salt Kimchi was prepared with salt replacements (NaCl 50%, KCl 36%, $MgSO_4$ 10%, $CaSO_4$ 3%, glutamic acid 0.2%). Chemical characteristics and microbiological parameters were monitored during fermentation at $20^{\circ}C$. When three kinds of preservative which were alcohol 2.0%, adipic acid 0.1% and their mixture were added to low-salt Kimchi, shelf-life of them were prolonged. To extend the shelf-life of low-salt Kimchi, when the heat treatment at $40^{\circ}C$ and $60^{\circ}C$ were tried, heat treatment at $60^{\circ}C$ was superior than at $40^{\circ}C$. In sensory evaluation of low-salt Kimchi, the control was showed the best quality in the overall acceptability. And low-salt Kimchi treated at $40^{\circ}C$ showed the most similar characteristics to the control Kimchi.
Kim, Byung-Hyuk;Jeong, Su Jin;Jang, Jong-Ok;Lee, Jun-Hyeong;Park, YeEun;Kim, Jung-Gyu;Kwon, Gi-Seok;Hwang, Hak-Soo;Lee, Jung-Bok
Journal of Life Science
/
v.29
no.8
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pp.846-853
/
2019
Phytochemical compounds of Ligusticum striatum Makino are used as traditional medicinal herbs in Asia. These compounds are reported to have pain relief and antioxidant activities in gynecological and brain diseases. In this study, we investigated the antioxidant effects of Ligusticum fermented ethanol extract from Lactobacillus plantarum BHN-LAB 129 isolated from Kimchi, a Korean traditional food. The total polyphenol and total flavonoid contents increased by about 116.2% and 281.0% respectively, in the fermented Ligusticum extract as compared with those in the nonfermented Ligusticum ethanol extract. Superoxide dismutase-like (SOD), DPPH radical scavenging, ABTS radical scavenging, and reducing power activities increased by around 139.9%, 199.6%, 301.0%, and 137.1%, respectively, in the fermented Ligusticum ethanol extract as compared with these parameters in the nonfermented Ligusticum ethanol extract, respectively. In conclusion, the fermented Ligusticum ethanol extract with L. plantarum BHN-LAB 129 was effective in increasing the antioxidant effects. The bioconversion process in this study points to the potential of using Ligusticum to produce phytochemical-enriched natural antioxidant agents with high added value. The findings may prove useful in the development of improved foods and cosmetic materials.
The aim of this study was to investigate the effects of Korean rice wine residue (RWR) on the growth performance and blood profiles of Hanwoo steers in the fattening stage. In situ and in vivo experiments were conducted to analyze rumen fermentation characteristics and total tract digestibility, respectively. Three cannulated Hanwoo steers (mean body weight: 448 ± 30 kg) were used in both analyses. The growth performance of 27 experimental animals in the fattening stage (initial body weight: 353.58 ± 9.76 kg) was evaluated after 13 months of feeding. The animals were divided into three treatment groups (n = 9/group). The treatments comprised total mixed ration (TMR) only (CON), TMR + 10% RWR (10% RWR), and TMR + 15% RWR (15% RWR). The diets of equal proportions were fed daily at 08:00 and 18:00 h based on 2% of the body weight. The animals had free access to water and trace mineral salts throughout the experiment. Supplementation of 15% RWR significantly decreased (p < 0.05) the rumen fluid pH compared with the control treatment, but there was no significant difference in the total volatile fatty acid concentration. It also significantly increased (p < 0.05) dry matter digestibility compared with the other treatments. The total weight gain and average daily gain of the animals in the RWR-supplemented groups were significantly higher (p < 0.05) than those in the control group. Furthermore, the feed intake and feed efficiency of the RWR-supplemented groups were higher than those of the control group. Supplementation of RWR did not affect the alcohol, albumin, glucose, total cholesterol, triglyceride, and low-density lipoprotein concentrations, and aspartate aminotransferase and alanine transaminase activities in the blood; these parameters were within the normal range. The high-density lipoprotein and creatinine concentrations were significantly higher in the 15% RWR group, whereas the blood urea nitrogen concentration was significantly higher in the 10% RWR group than in the other groups. These results suggest that TMR with 15% RWR can serve as an alternate feed resource for ruminants.
This study was conducted to investigate the effects of dietary supplementation of CS682, a fermentation product of Actinomycetae Nocardia sp. CS682, and DCS682$^{(R)}$, a commercial product, on the performance, blood parameters, small intestinal microflora, and immunoglobulin contents in broilers. In Exp. 1, a total of 240 ROSS$^{(R)}$ broiler chickens of 1d old were assigned to six dietary treatments: Control, Antibiotics (6 ppm avilamycin), CS682-0.25 (CS682 0.25%), CS682-0.50, CS682-0.75 and CS682-1.00. There were significant (p<0.05) differences among treatments in feed conversion. The CS682-0.25 treatment was significantly (p<0.05) lower than Antibiotics and other CS682 treatments in 0~2 wk feed conversion. The CS682 treatments influenced MCV (mean corpuscular volume) in blood. The cfu of Escherichia coli in small intestinal content was lowest in Antibiotics treatment followed by CS682 treatments and Control. In Exp. 2, a total of 1,000 ROSS$^{(R)}$ broiler chickens of 1 d old were assigned to five dietary treatments: Control, Antibiotics (6 ppm avilamycin), DCS682-0.05 (DCS682$^{(R)}$ 0.05%), DCS682-0.10 and DCS682-0.20. There were significant differences (p<0.05) among treatments in mortality. The DCS682-0.20 treatment was lower than DCS682-0.10 in 0~3 wk and lower than Control in 0~5 wk mortality. Antibiotics treatment was lowest in all microbial population in small intestinal content. The cfu of E. coli and Salmonella typhimurium of DCS682 treatments were higher than Antibiotics treatment but lower than the Control. The results of present broiler experiments indicated that supplementation of 0.20~0.25% CS682 and DCS682, improve feed conversion, mortality and control harmful intestinal microbes.
Ku, Hwa-Suk;Noh, Jeong-Sook;Yun, Ye-Rang;Kim, Hyun-Ju;Kwon, Myung-Ja;Cheigh, Hong-Sik;Song, Yeong-Ok
Journal of the Korean Society of Food Science and Nutrition
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v.36
no.9
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pp.1140-1147
/
2007
A recipe for diet kimchi containing 20% of sea tangle to Korean cabbage kimchi (SK) was developed for weight reduction purpose. The fermentation process of SK showed typical Lactobacillus sp. growth pattern like other kimchis. The effects of SK on weight reduction was studied in high fat diet (HFD) fed rats (n=40). Diet groups used for the animal study were HFD, HFD supplemented either with Korean cabbage kimchi used as experimental control (HCK), or with SK (HSK), or with J-kimchi (JK) which was purchased at market (FJK). The effect of kimchi on preventing obesity in rat fed HFD was found to be obvious by means of reducing visceral fat contents and improving serum lipid profiles through enhancing the lipid excretion in the feces (p<0.05). Leptin concentration of rat was significantly decreased by kimchi consumption (p<0.05). This result can be interpreted that adipocytes in these animals were fewer than that of HFD group. The plasma bililubin concentration was lower in kimchi group than HFD, meaning that returning bile from ileum to the liver was reduced. When the observe beneficial effects of kimchi on preventing obesity were compared among kimchi groups, SK only reduced the relative visceral fat contents significantly than other kimchi groups (p<0.05). Besides this, other parameters such as plasma lipid profiles, feces lipids, leptin, and bililubin concentration were not significantly different, even though the most beneficial effect on these parameters was observed from SK. In conclusion, long term consumption of SK seems to have a beneficial effect on the prevention of obesity through enhancing the excretion of lipids in the feces. The dietary fiber content of SK was increased by 7% compared to CK when 20% of sea tangle was added.
This study was conducted to investigate the effects of dietary supplementation of CS682, a fermentation product of Actinomycetae(Nocardia sp. CS682), and its commercial product DSC682$^{(R)}$ on the performance, blood parameters, intestinal microflora, and immune response in laying hens. Hy-Line Brown$^{(R)}$ laying hens were housed in two bird cages. Feeding trial lasted 5 wk under 16.5 h:7.5 h(L:D) lighting regimen. In Exp.1, a total of 480 birds of 86 wk old were assigned to four dietary treatments: Control, Antibiotics (6 ppm avilamycin), CS682-0.1 (CS682 0.1%) and CS682-1.0 (CS682 1.0% supplementation). Each treatment was replicated five times with 24 birds (or 12 cages) per replication. In Exp. 2, a total of 1,000 birds of 26 wk old were assigned to five dietary treatments: Control, Antibiotics (6 ppm avilamycin), DCS682-0.05 (DCS682 0.05%), DCS682-0.1 (DCS682 0.1%), DCS682-0.2 (DCS682 0.2% supplementation). Each treatment was replicated five times with 40 birds (or 20 cages) per replication. In Exp. 1, there were no significant differences among treatments in egg production, egg weight, broken & soft egg production, feed intake, and feed conversion ratio. Also, there were no significant differences among treatments in eggshell thickness, eggshell color and Haugh unit. However, eggshell strength was significantly (p<0.05) greater in CS682 and Antibiotics treatments than Control, and egg yolk color was significantly (p<0.05) higher in CS682-1.0 than Control. In Exp. 2, feed intake was significantly (p<0.05) lower in DSC682-0.05 than Control. Lightness(L) of Hunter Lab color of eggshell of DCS and Antibiotics treatments was significantly (p<0.05) lower than Control. Egg yolk color of DCS 0.1 and 0.2 treatments was significantly (p<0.05) higher than Control. Haugh unit increased significantly (p<0.05) in Antibiotics and DCS682-0.1 treatments. The immunoglobulin levels of plasma (IgG and IgA) and eggyolk (IgY) were not significantly affected by treatments. Antibiotics and CS682 or DCS682 treatments significantly (p<0.05 or 0.01) influenced some of the erythrocytes and leukocytes parameters in blood. In Exp.1, mean corpuscular volume (MCV) decreased by CS682 treatments and mean corpuscular hemoglobin (MCH) was highest in Antibiotics treatments. In Exp.2, the level of monocyte (MO) decreased in DCS682-0.10 and 0.20 treatments. The cfu of C. perfringens and S. typhimurium in small intestinal content were highest in Control and lowest in Antibiotics in both experiments. In Exp. 2, DSC682-0.05 and -0.1 treatments were highest and Antibiotic treatment was lowest in Lactobacilli spp. The results of the present layer experiments indicated that supplementation of 0.1~0.2% CS682 or DCS682 may increase eggshell strength, color of eggshell and eggyolk, Haugh unit, and control harmful intestinal microbes.
Kim, Y.I.;Jun, S.H.;Yang, S.Y.;Huh, J.W;Kwak, W.S.
Journal of Animal Science and Technology
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v.49
no.5
/
pp.667-676
/
2007
This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.
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