• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.197-203
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• 2009

An indole oxygenase originated from Rhodococcus sp. RHA1 was cloned into the expression vector, pTrc99A, in Escherichia coli, and designated pTCAN1. The pTCAN2 was constructed from pTCAN1 by the deletion of $lacI^q$ for the constitutive expression of indole oxygenase without adding IPTG in the medium. The complete open reading frame of indole oxygenase was 1,224 bp long, which encodes a protein of 407amino acids. Crude extracts of E. coli $DH5{\alpha}$/pTCAN1 and pTCAN2, respectively, were prepared and subjected to SDS-PAGE analysis. A band corresponding to molecular mass of about 43 kDa was appeared and this result correlated with the predicted molecular mass of cloned indole oxygenase. The E. coli harboring pTCAN1 and pTCAN2, respectively, showed blue color colony in LB plate. The pigment showing blue color was prepared from E. coli $DH5{\alpha}$/pTCAN2, and identified as indigo by experiments using spectrophotometer, HPLC, and TLC. The indigo-forming activity of indole oxygenases from the whole cell of E. coli $DH5{\alpha}/pTCAN1$ cultured at LB medium added 1mM of IPTG and that of E. coli/pTCAN2 showed about 1.75nmol/min/mg DCW (dry cell weight) and 3.85 nmol/min/mg DCW, respectively. Also, the E. coli $DH5{\alpha}$/pTCAN2 produced about $236{\mu}M$ of indigo after 48 hours incubation in TB medium supplemented with 2.5 mM of tryptophan.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.64-69
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• 1991

- The effects of sodium acetate on the production of $\gamma$-linolenic acid (GLA) and the secretion of the mycelial lipid into the culture medium with noninnic surfactants were studied with Mucor sp. KCTC 8405P. In the addition of 2.0%) sodium acetate to the basal medium, dry cell weight and total lipid content were increased from 7.8 g/l and 2.46 g/l to 16.0 g/1 and 4.77 g/l, but GLA content was decreased from 18.6% to 13.85%. The growth of Mucor sp. KCTC 8405P was greatly dependent on both the initial pH and the concentration of sodium acetate of culture medium, which was considered as the results of the formation of acetic acid because the fungal growth was completely inhibited at the concentration of acetic acid higher than 22 mM. With the decrease of the oxygen supply, the cell growth, total lipid, and GLA content were sharply decreased in the presence of 2.0% sodium acetate. For the secretion of mycelial lipid into the culture medium, the effects of the various nonionic surfactants were examined. In the addition of 0.5% Tween 80 or Span 80 to the basal medium, 194 mg/l or 263 mg/l of GLA was obtained in the culture medium.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.12
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• pp.1732-1737
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• 2003

An experiment was conducted to investigate the effect of feeding transgenic cottonseed (Bt.) vis-a-vis non-transgenic (non-Bt.) cottonseed on blood biochemical constituents in lactating Murrah buffaloes. Twenty Murrah buffaloes in mid-lactation were divided into 2 groups of 10 each. Animals of group I were fed with 39.5% non-transgenic cottonseed in concentrate mixture while the same percentage of transgenic (Bt.) cottonseed was included in the concentrate mixture fed to the animals of group II. Animals of both groups were fed with concentrate mixture to support their milk production requirements. Each buffalo was also offered 20 kg mixed green fodder (oats and berseem) and wheat straw ad libitum. The experimental feeding trial lasted for 35 days. There was no significant difference in the dry matter intake between the two groups of buffaloes. All the buffaloes gained body weight, however, the differences were non significant. Total erythrocyte count, hemoglobin content and packed cell volume were $9.27{\pm}0.70${\times}10^6/{\mu}l$, $13.01{\pm}0.60gdl$ and $34.87{\pm}1.47%$, respectively in group I with the corresponding figures of $8.88{\pm}0.33$, $12.99{\pm}0.52$ and $31.08{\pm}1.52$ in group II. The values of total erythrocyte count, haemoglobin content and packed cell volume did not differ significantly between the two groups of buffaloes. The concentration of plasma glucose, serum total proteins, albumin, globulin, triglycerides and high density lipoprotein were non significantly higher in buffaloes fed non-transgenic cottonseed than in buffaloes fed transgenic cottonseed. The cholesterol concentration was significantly (p<0.01) higher in buffaloes of group I ($136.84{\pm}8.40mg/dl$) than in buffaloes of group II ($105.20{\pm}1.85mg/dl$). The serum alkaline phosphotase, glutamic-oxaloacetate transaminase and glutamic-pyruate transaminase activities did not differ significantly between two groups of buffaloes. However, serum glutamic-pyruate transaminase activity was considerably high in buffaloes fed nontransgenic cottonseed as compared to buffaloes fed transgenic cottonseed. Bt. proteins in serum samples of animals of group II were not detected after 35 days of feeding trial. It was concluded that transgenic cottonseed and non-transgenic cottonseed have similar nutritional value without any adverse effects on health status of buffaloes as assessed from haematobiochemical constituents.

• Asian-Australasian Journal of Animal Sciences
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• v.8 no.4
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• pp.393-402
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• 1995

Experiments were conducted to evaluate the nutritive values of supplemental L-lysine, liquid and powder type, and DL-methionine in weanling pigs. For feeding trial, 165 weanling pigs were treated in 2 controls; 18 and 16% CP, 6 supplementations of lysine alone to 16% CP diets; 0.1, 0.2 and 0.4% of liquid and powder type each, and 3 supplementations of lysine + methionine to 15% CP diets; 0.05 + 0.025, 0.1 + 0.05 and 0.2 + 0.1%. Pigs were fed for 5 week to investigate the protein sparing effect of supplemental amino acid, and the optimal supplemental level. A metabolic trial included the measurements of digestibilities of dry matter, crude protein, crude fat, crude fiber, energy, phosphorus and amino acids. The liver acinar cell culture was conducted for the protein synthesis activity of the pigs fed each experimental diet. Supplementation of both type of L-lysine in 16% CP diet showed improved daily weight gain and feed efficiency which were compatible with those of pigs fed 18% CP diet. Groups fed liquid lysine did not differ from those fed powder type in growth performance. Supplementation of lysine and methionine to 15% CP diet did not improve growth performance of pigs to the extent that 18% CP diet was fed. In nutrient digestibility, 16% CP control diet showed significantly (p < 0.05) lower crude protein digestibility than any other treatments. Digestibilities of 16% CP diets with lysine supplementation were equal to that of 18% CP control, while digestibilities of 15% CP diets with the supplementation of lysine + methionine was inferior to that of 18% CP control. Supplementation of lysine alone reduced the nitrogen excretion compared to the none supplemented control groups. However, addition of lysine + methionine excreted more nitrogen than controls. Pigs fed diet supplemented with lysine alone, or lysine + methionine excreted less fecal phosphorus than those fed none supplemetation. Retained protein from liver tissue of pigs fed 18% diet was significantly (p < 0.05) greater than those fed 16% CP diet. A significant difference (p < 0.05) was observed in physical type of lysine. Feeding of powder type showed less secreted protein and greater retained protein in the culture of liver acinar cell. It is concluded that supplementation of lysine at the level of 0.1 to 0.2% can spare 2% of dietary protein and reduce nitrogen excretion by 19.3%. Also, no difference in nutritional values was observed between liquid and powder lysine in weanling pigs.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.7
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• pp.998-1003
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• 2016

Although exogenous protease enzymes have been used in poultry diets quite extensively, this has not been the case for pig diets. In general, due to their better gut fermentative capacity and longer transit time, pigs have greater capacity to digest dietary proteins than poultry. However, in early-weaned piglets, the stress brought about by weaning adversely affects the digestion of dietary proteins. Therefore, a study was conducted to determine the effects of a commercial protease enzyme in weanling pigs. Indices of growth, nutrient digestibility, blood profiles, fecal microflora, fecal gas emission and fecal scores were measured during the study. A total of 50 weanling pigs ($6.42{\pm}0.12kg$) at 28 d of age were randomly assigned to receive 1 of 2 dietary treatments: i) control diet (corn-soy based) with no supplemental protease (CON), and ii) control diet+200 g/ton protease (PROT) for 42 d. A completely randomized design consisting of 2 treatments, 5 replicates, and 5 pigs in each replicate was used. Growth performance in terms of body weight ($27.04{\pm}0.38kg$ vs $25.75{\pm}0.39kg$; p<0.05) and average daily gain ($491{\pm}7.40g$ vs $460{\pm}7.46g$; p<0.05) in PROT fed pigs were increased significantly, but gain per feed ($0.700{\pm}0.01$ vs $0.678{\pm}0.01$; p>0.05) was similar between treatments at d 42. Relative to CON pigs, PROT fed pigs had increased (p<0.05) apparent total tract digestibility ($84.66%{\pm}0.65%$ vs $81.21%{\pm}1.13%$ dry matter and $84.02%{\pm}0.52%$ vs $80.47%{\pm}1.22%$ nitrogen) and decreased (p<0.05) $NH_3$ emission ($2.0{\pm}0.16ppm$ vs $1.2{\pm}0.12ppm$) in the feces at d 42. Except for a decreased (p<0.05) in blood creatinine level, no differences were observed in red blood cell, white blood cell, lymphocyte, urea nitrogen, and IgG concentrations between treatments. Fecal score and fecal microflora (Lactobacillus and E. coli) were also similar between CON and PROT groups. Overall, the supplementation of protease enzyme in weanling pigs resulted in improved growth rate and nutrient digestibility. Exogenous protease enzyme reduced fecal $NH_3$ emission, thus, potentially serving as a tool in lowering noxious gas contribution of livestock production in the environment.

• Korean Journal of Microbiology
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• v.37 no.1
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• pp.92-99
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• 2001

High-cell-density cultivation of Ralstonia eutopha KHB 8862 by fed-batch fermentation in a 200 l pilot plant was carried out for the mass production of poly(3-hydroxybutyrate) (PHB). After 80 h of cultivation, the dry cell weight (DCW), PHB concentration, and PHB yield from fructose syrup reached 168 g/l, 74%DCW, and 0.27 (w/w), respectively, resulting in a productivity of 1.6 g of PHB/L/h. Based on these results, the PHB production cost from bacterial fermentation was analyzed and economic evaluation was performed. In the case of new investment being implemented or not, the production cost of PHB was US$ 3.15/kg and US$ 2.41/kg, respectively. PHB productivity and PHB yield on a carbon substrate were both important factors to be optimized. The increase of PHB yield on a carbon sources significantly decreased the PHB production cost but the increase in productivity had a relatively slight effect on the decrease in PHB production cost because the cost of carbon sources (37%) for PHB was larger in proportion to total cost than the depreciation cost (17%). These results suggest that the increased PHB yield from carbon sources and the development of new cheaper substrates would be more effective in decreasing PHB production cost than the increase in productivity. It was demonstrated that PHB is not in competition with consumable plastics such as PET in present market. Therefore, it is essential to lower production cost to be used as a bulk product and desirable to develop new application fields for PHB such as biomedical and cosmeceuticals.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.556-560
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• 1997

The removal efficiency of COD and the production of ${\delta}-aminolevulinic$ acid (ALA) were concurrently investigated for both purifying the soybean curd wastewater of high BOD and utilizing the wastewater as a renewable substrate of ALA production using Rhodobacter capsulatus KK-10. Its wastewater was a favorable media for the growth of photosynthetic bacteria in terms of its environmental characteristics having COD/BOD rate of 0.98, ratio of BOD : N : P=100 : 6 : 4, BOD/N ratio of 17.2, lactic acid of 1,080 ppm. Its COD value wastewater was decreased to 94% and dry cell weight was approached to about 1.2 g/l after cultivation of the photosynthetic bacteria for 4 days. By the addition of 15 mM levulinic acid (LA) into the wastewater at the middle log phase of cell growth, the amount of ALA secreted was 55 mg/l. The ALA production was considerably increased to 114 mg/l under the cultural condition of 15 mM supplementations of glycine and succinate with LA at the same period. Furthermore the maximum ALA production of 120 mg/l and COD removal efficiency of 92% were accomplished in the soybean curd wastewater enriched with one addition of 15 mM LA and three serial additions 15 mM ALA precursors.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.337-343
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• 2005

Tapioca alcoholic distillery waste was utilized as dual purposes to produce citric acid and to reduce the amount of waste to be treated. Primarily an attempt was made to optimize the process conditions by Aspergillus niger in shake bath. The effects of pH, temperature, nitrogen and phosphorus sources on citric acid production were investigated. Maximum concentration of citric acid was made at temperature of $30^{\circ}C$ and pH of 4.3, while maximum cell dry weight was obtained at $35^{\circ}C$. The addition of methanol or ethanol to culture medium promoted citric acid production remarkably, but the addition of $NH_4NO_3,\;KH_2PO_4$ and Manganese as mineral source decreased the acid production.

• Restorative Dentistry and Endodontics
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• v.28 no.5
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• pp.385-391
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• 2003

The purpose of this study is to examine the viability of PDL cells in rat molars by using MTT assay and to verify the MTT assay through the histologic observation. Thirty of Sprague-Dawley white female rats of 4-weeks old with a body weight of about 100 grams were used. Groupings are as follows : Immediate Group : Positive control group(n=10)-after extraction immediately. Dried Group : Negative control group(n=10)-after drying for an hour under warm dry. $ViaSpan^{\circledR}$ Group : 1hour $ViaSpan^{\circledR}$ group(n=10)-after storing in $ViaSpan^{\circledR}{\;}at{\;}4^{\circ}C$ for 1hour. Ten teeth of each group were treated as same as above and replanted to the original socket of experimental animals. After two weeks of replantation. all the experimental animals were sacrificed. And after fixation, extracted maxillary jaw was dimineralized. After it was embedded in paraffin. serial section by $5\mu\textrm{m}$ was carried out and for construction of specimen, hematoxylin-eosin dye was used. The mean MTT measurement of immediate group(positive control) is 2.81 and the mean measurement of dried group(negative control) is 0.98 which is significantt differnt(P<0.05), The mean measurement of $ViaSpan^{\circledR}$ group is 2.65 and there is significant difference between dried group and $ViaSpan^{\circledR}$ group(P<0.05), However, there is no difference between immediate group and $ViaSpan^{\circledR}$ group. The average resorption points of immediate group is 3.03 points. In the dried group, average 6.44 points resorption and 2.68 points showed resorption in the $ViaSpan^{\circledR}$ group. Unlike with MTT assay, there was no significant difference between the immediate group and $ViaSpan^{\circledR}$ group. The usage of MTT assay as a viable cell marker may give us a better indication of the maintenance of periodontal ligament cell vitality.

• Restorative Dentistry and Endodontics
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• v.31 no.5
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• pp.398-408
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• 2006

The purpose of this study was to verify the usefulness of MTT analysis as a tool of measurement of the periodontal ligament cell viability from the extracted rat molar. A total of 80 Sprague-Dawley white female rat of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted under Ketamine anesthesia. Twenty-four teeth of each group (divided as five groups depending upon the time-lapse after extraction such as Immediate, 10, 20, 40 and 60 minutes) were immersed in $200{\mu}l$ of MTT solution (0.5 mg/ml) and processed for optical density measurements. Another 10 teeth of each group were treated as same as above and sectioned at $10{\mu}m$ for microscopic examination. All measurements values were divided by the value of hematoxylin-eosin staining which represented the volume of each corresponding samples. Immediate and 10 minute groups showed highest MTT values followed by 20, 40, and 60 minutes consecutively. Statistical significance (p<0.05) existed between all groups except in immediate versus 10 minute groups and 40 versus 60 minutes. Histological findings also showed similar findings with MTT results in crystal shape and crystal numbers between the experimental groups. These data indicate that in vivo MTT analysis nay be of value for evaluation of the periodontal ligament cell viability without time- consuming cell culturing processes.