• Title/Summary/Keyword: K11 lysozyme

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Fouling Study with Binary Protein Mixtures in Microfilration (이성분계 단백질 혼합물의 미세막 분리공정에서 막오염에 관한 연구)

  • Ahn, Byung Hun;Moon, Dong Ju;Yoo, Kye Sang;Ho, Chia Chi
    • Korean Chemical Engineering Research
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    • v.43 no.2
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    • pp.236-242
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    • 2005
  • Membrane fouling by protein mixtures during microfiltration has been investigated for binary mixtures of bovine serum albumin (BSA), casein, lysozyme, pepsin, and ovalbumin. Filtration experiments were carried out using $0.2{\mu}m$ polycarbonate track-etched (PCTE) membrane in a stirred cell under constant transmembrane pressure (14 kPa) and concentration of hydrogen ion (pH=11) to study the effect of mixture composition on filtrate flux decline. Flux decline data were analyzed using a pore blockage-cake formation model developed recently. It was found that the model is in a good agreement with the experimental data. Fouling parameters such as the rate of pore blockage(${\alpha}$), the initial resistance of the protein deposit ($R_{po}$) and the increasing rate of the protein layer resistance(${\beta}$) were used to evaluate the rate of filtrate flow by membrane fouling in the binary mixture system. Generally, the trend of ${\alpha}$ is comparable with that of filtrate flux decline. It was also found that fast flux decreasing was observed over the binary mixture containing casein. The result is due to high value of the initial resistance of the protein deposit ($R_{po}$) over casein.

Characterization of Bacteriocin Production by Lactococcus lactis LAB3113 Isolated from Kimchi

  • Shin, Jong-Yeun;Cheol Ahn
    • Preventive Nutrition and Food Science
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    • v.2 no.2
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    • pp.101-108
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    • 1997
  • A lactic acid bacterium LAB3113, isolated from traditionally fermented Kimchi was found to produce bacteriocin whose activity was very specific toward lactobacilli and not effective against any other bacteria. Lactobacilli affected by the inhibitory substance included Lactobacillus delbrueckii-lactis, L. johnsonii, L. gsseri, and L. curvatus. Based upon biochemical and physiological characteristics, LAB3113 was classified as Lactococcus lactis, and its bacteriocin was named as lactococcin K3113. Lactococcus lactis. LAB3113 produced bacteriocin at th early stage of growth and the concentration of the bacteriocin did not decrease even after alt stationalry phase. Optimal temperature of bacteriocin production was $25^{\circ}C$ at the initial pH 7.0. Partially purified lactococcin K3113 was completely inactivated by protease, but not affected by lipase, lysozyme and RNase. The bacteriocin was very heat-stable even after autoclaving for 20 min. It was also stable in pH changes, an was not affected by th presence of solvents. lacotococcin K3113 appeared to act in bactericidal mode against L. delbrueckii-lactis ATCC4797. Molecular weight of lactococcin K3113 was calibrated as 10,500 dal by SDS-PAGE an activity staining. Lactococcus lactis LAB3113 had four residential plasmids of 3.7kb, 11.2kb, 15.5kb, and 48kh in molecular sizes. Plasmid profile analysis of mutant strain revealed that 15.5 kb plasmid was re-sponsible for the production of lactococcin K3113 and its immunity to the bacteriocin.

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Characterization of the Cell-Surface Barriers to Plasmid Transformation in Corynebacterium glutamicum

  • Jang, Ki-Hyo;Paul J. Chambers;Chun, Uck-Han;Margare L.Britz
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.294-301
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    • 2001
  • The effects of including glycine and isonicotinic acid hydrazide (INH) in the growth medium (Luria broth, LBG) on the subsequent lysozyme-imduced protoplast formation and transformation efficiency of Corynebacterium glutamicum were studied. The transformation efficiency of C. glutamicum AS019 increased up to 100-fold as the ocncentrationof glycine in the media increased from 0% to 5% (w/v), relative to cells grown in the absence of glycine. The presence of 5 mg/ml INH in the growth medium led to a further 10-fold increase in transformation efficiency. In addition, this transformation protocol was successfully applied to other strains of C. glutamicum. Both chemicals affected the mycolic acid attachment to the cell surface of C. glutamicum, when INH, the relative percentage of fatty acids of AS019 to the total lipids (mycolic acid plus fatty acids) decreased from 76.9% (in LBG) to 72.9% (in LBG-2% glycine) and 66.4% (in LBG-8 mg InG/ml), thereby suggeting that these chemicals also inhibit fatty acid synthesis.

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Effects of Dietary Probiotics Supplementation on Juvenile Korean Rockfish Sebastes schlegeli (치어기 조피볼락 Sebastes schlegeli 사료내 생균제 첨가효과)

  • Lee, Seung-Hyung;Yoo, Gwang-Yeol;Choi, Se-Min;Kim, Kang-Woong;Kang, Yong-Jin;Bai, Sung-Chul C.
    • Journal of Aquaculture
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    • v.21 no.2
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    • pp.82-88
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    • 2008
  • A 12-week feeding trial was conducted to investigate the effects of dietary supplementation of probiotics as a feed additive for juvenile Korean rockfish Sebastes schlegeli. Four experimental diets supplemented with no probiotic(Control), Bacillus polyfermenticus(BP), Bacillus licheniformis(BL) or Bacillus polyfermenticus plus Saccharomyces cerevisiae(BP+SC) at $1.0{\times}10^7$ CFU/kg diet as a dry-mater(DM) basis were prepared by mixing with a basal diet. After 12 weeks of the feeding trial, fish fed BP+SC diet showed significantly higher weight gain(WG), feed efficiency(FE), specific growth rate(SGR) and protein efficiency ratio(PER) than those of fish fed control diet(P<0.05), however there were no significant differences in WG, FE, SGR and PER among fish fed the BP, BL and BP+SC diets. Fish fed BP and BP+SC diets showed significantly higher condition factor(CF) than that of fish fed control and BL diets. Fish fed BP, BL, BP+SC diets showed significantly higher hepatosomatic index(HSI) than that of fish fed control diet, however there was no significant difference in HSI among fish fed BP, BL and BP+SC diets. Fish fed BP+SC diet showed significantly lower serum glucose than that of fish fed control diet, however there was no significant difference in serum glucose among fish fed BP, BL and BP+SC diets. Fish fed BP+SC diet showed significantly higher respiratory burst activity(NBT assay) than that of the fish fed control and BL diets, however there was no significant difference in NBT assay between fish fed BP and BP+SC diets. Fish fed BP and BL diets showed significantly higher lysozyme activity than that of the fish fed control diet, however there was no significant difference in lysozyme activity among fish fed BP, BL and BP+SC diets. Fish fed BP and BP+SC diets showed significantly lower cumulative mortality than that of the fish fed control diet, however there was no significant difference in cumulative mortality among fish fed BP, BL and BP+SC diets after the challenge test. From these results, dietary B. polyfermenticus, B. licheniformis and B. polyfermenticus plus S. cerevisiae supplementation in juvenile Korean rockfish diet could enhance growth performances, non-speicific immunities and a higher resistance against the specific pathogen.

Dietary inclusion of mealworm (Tenebrio molitor) meal as an alternative protein source in practical diets for rainbow trout (Oncorhynchus mykiss) fry

  • Jeong, Seong-Mok;Khosravi, Sanaz;Mauliasari, Intan Rizki;Lee, Sang-Min
    • Fisheries and Aquatic Sciences
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    • v.23 no.5
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    • pp.12.1-12.8
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    • 2020
  • Background: An 8-week feeding trial was designed to evaluate the potential of yellow mealworm (MW; Tenebrio molitor) as a locally available nutrient-rich feedstuff for rainbow trout fry (Oncorhynchus mykiss). Methods: Triplicate groups of fish (mean ± SE; 1.11 ± 0.01 g) were assigned to each of the five isonitrogenous and isocaloric practical diets containing graded level of a full fat MW (0, 7, 14, 21, and 28%) at the expense of fish meal (designated as MW0, MW7, MW14, MW21, and MW28, respectively). Results: Fish growth performance in terms of weight gain and specific growth rate significantly increased with increasing dietary MW level up to 14% and then declined when dietary MW levels further increased to 28%. Significantly higher protein efficiency ratio and lower feed conversion ratio were found in fish fed with diets containing MW compared to fish fed the control MW0. Myeloperoxidase activity was significantly higher in fish fed MW7 diet compared to fish fed the MW0 diet. Fish fed the MW14 and MW28 diets had significantly higher lysozyme activities than those fed the MW0 diet. Conclusions: Overall, the efficacy of MW as promising alternative to fish meal in practical diets for rainbow trout fry has been proved not only in relation to growth rates and feed utilization, but also from the viewpoint of immunopotentiation effects.

Transformation Conditions of Bacillus subtilis by Streptomyces rimosus Plasmid DNA (Streptomyces rimosus Plasmid DNA에 의한 Bacillus subtilis의 형질전환 조건)

  • Hong, Yong-Ki;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.75-79
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    • 1983
  • To exploit a suitable vector and recipient strain for molecular cloning in Bacillus subtilis, oxytetracycline-resistant plasmic DNA has been prepared from Streptomyces rimosus by phenol-buffer extraction of lysozyme-lysed cells and introduced into B. subtilis KPM 60 [St $r^{R}$-mutant of RM 125 (leu A8, arg 15, hsm $M^{-10}$ , hsr $M^{-10}$ )] by transformation. Oxitetracycline-resistant plasmid was well transferred into B. subtilis KPM 60 with average frequency of 10$^{-4}$ per $\mu\textrm{g}$ of DNA. The highest frequency of plasmid transformation was obtained after 3 hours incubation of recipient cells in the growth medium and 30 to 60 minutes incubation in the competence medium, and then 20 minutes contact of DNA and host cells. Optimum pH for competence was 7.5, and optimum temperature for transformation was 2$0^{\circ}C$.>.

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Comparison of Feed Efficiency Between Rotifers Enriched Lipid-contents to Enrichment and Enhanced Digestive Enzymes Activity to Starch (영양강화 Rotifer와 효소활성 향상 Rotifer의 먹이효율 비교)

  • Kwon, O-Nam;Park, Heum-Gi
    • Journal of Aquaculture
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    • v.22 no.1
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    • pp.105-111
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    • 2009
  • In this study, we carried out an experiment for estimation the larval digestibility in aspects which digestive enzymatic activities and nutrition of the rotifers, Brachionus rotundiformis. Thus we enhanced the digestive enzymatic activity through the addition of starch for the increase of digestibility of rotifer (starch-rotifer), and compared with the feed efficiency through rearing of the olive flounder, Paralichthys olivaceus used rotifer lipid-enriched with Algamac $2000^{(R)}$ (CE-rotifer). The digestive enzyme activities (except for TG-lipase), total protein contents, total essential amino acid, essential amino acids (methionin and phenylalanine) of starch-rotifer (the rotifer used a starch as additive, and enriched not) was assayed significantly higher than CE-rotifer (P<0.05). And total lipid, lipid classes (except for sterol) and fatty acids as DHA and EPA showed higher in CE-rotifer than starch-rotifer (P<0.05). But, sterol contents and ST/TG ratio were shown significantly higher in starch-rotifer (P<0.05). The flounder larvae supplied the two rotifers showed standard length and body weight that not significantly differed with ranges $3.72{\sim}3.79\;mm$ and $32.9{\sim}37.8\;mg$/larva on 6 days after hatching (DAH), respectively (P>0.05). However, these of 12 DAH showed the values of significantly higher to $5.94{\pm}0.249\;mm$, $144.0{\pm}23.86\;mg$/larva and $26.2{\pm}12.13%$ in standard length, body weight and survival in CE-flounder than that of starch-flounder (P<0.05). The hydrolytic enzymatic activities of flounder larvae severally supplied the two rotifers showed the significantly higher activities in acidic -amylase, neutral -amylase, TG-lipase, lysozyme and acidic phosphatase in starch-flounder on 5 DAH (P<0.05). But neutral $\alpha$-amylase, three proteases and two phosphatases of CE-flounder on 11 DAH showed the significantly higher activities than that of starch-flounder (P<0.05). Therefore, for the flounder, Paralichthys olivaceus larvae just depleted yolk was more beneficial to supply the feed, rotifer, enhanced the digestibility than to supply the feed lipid-enriched for aspect of larval digestibility up to 6 DAH, thereafter nutrition of absorption due to the development of digestive organs suggested that enrichment effect appeared with larval somatic growth. Consequently, investigation more detailed about the larval digestive physiological and nutritional requirement variations after 6 DAH will be necessary, thereafter.

Dietary Administration of Probiotics, Bacillus sp. IS-2, Enhance the Innate Immune Response and Disease Resistance of Paralichthys olivaceus against Streptococcus iniae (Probiotics를 양식넙치에 투여시 Streptococcus iniae에 대한 면역반응 및 병저항성)

  • Jang, Ik-Soo;Kim, Dong-Hwi;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.172-178
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    • 2013
  • The strains were added to the feed in the concentration of $10^3$, $10^5$, and $10^7$ CFU/kg and 2% of fishes were given the feed twice a day (8 AM and 5 PM) for 12 weeks. In result of the nonspecific immune response study to examine Respiratory burst activity, Lysozyme activity and Phagocytosis activity every two weeks until the end of the study, all test samples showed greater activities than control samples and improved immune activity with Bacillus sp. IS-2. The mortality test performed by artificial infection using Streptococcus iniae, a pathogenic bacterium, after the completion of this study also showed over 55% greater survival rate in all test samples. In result of performing PCR using the universal primer to verify that the probiotic stays in the intestines of the fishes, all test samples showed PCR product of 1,465 bp. Based on the above findings, it was concluded that Bacillus sp. IS-2 in the feed improved farmed flatfish's immune system and resistance against diseases as the probiotics. Also, the physiological indicators discovered by this study would be useful for identifying the mechanisms of probiotics.

Immunocytochemical Characteristics of the Short-term Cultured Mesothelial Cells (단기배양한 중피세포의 면역세포화학적 연구)

  • Jeon, Ho-Jong;Lee, Mi-Ja;Lee, Mi-Sook;Jeong, Yu-Kyung;Lee, Young-Mi;Choi, Hyung-Ho
    • The Korean Journal of Cytopathology
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    • v.6 no.2
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    • pp.106-115
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    • 1995
  • Reactive humsn mesothelial cells were examined by immunocytochemical stain with intermediate filaments (cytokeratin [CK1, CK7, CK8, CK18, CD19), vimentin, desmin, actin), epithelial membrane antigen, carcinoembryonic antigen (CEA), MHC class II antigen (HLA-DR), LeuM-1 (CD15), $\alpha1-antitrypsin$(ACT), $\alpha1-antichymotrypsin$ (ACHT), CD68(KP-1) and FcyRIII(CD16). The mesothelial cells were isolated from patients with liver cirrhosis and pleural effusion, and short-term cultured in RPMI 1640 media containing 10% heat inactivated fetal calf serum and 1% identical supernatant fluid of the patients' transudates. The results obtained are as follows 1. The cultured-reactive mesothelial cells were positive for the protein of cytoskeleton such as cytokeratin and vimentin, but negative for desmin and actin. The resting mesothelial cells showed positive reactions for cylokeratin, but negative for vimentin, desmin and actin. 2. The primary antibodies to the cytokeratin were strongly reactive for CK1, CK8 and CK18 but negative for CK7 and CK19 in both reactive and resting mesothelial cells. 3. Resting mesothelial cells showed negative reactions for CEA, but strong positive reactions in cultured-reactive mesothelial cells. 4. The markers for the monocytes/histiocytes(CD11b, CD14, CD16, CD68, Iysozyme and $\alpha1-antitrypsin$ and $\alpha1-antichymotrypsin$) were nonreactive in resting mesothelial cells, but lysozyme and $\alpha1-antitrypsin$ were weakly reactive in reactive and proliferative mesothelial cells. 5. MHC Class II molecule(HLA-DR antigen) was negative in both resting and reactive mesothelial cells. These results suggest that the short-term cultured, reactive mesothelial cells show a newly aberrant expression of the vimentin and calcine-embryonic antigen. The reason of the aberrant expression of the intermediate filament and oncofetal antigen in reactive and proliferative mesothelial cells should be further evaluated.

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Patterns of Protein Leaching to Dispersion Medium during W/O/W Double Emulsion-Based Microencapsulation Processes (이중유제법에 근거한 미립자 제조 공정 중 단백질의 분산매로의 전이 양상)

  • Cho, Mi-Hyun;Choi, Soo-Kyoung;Sah, Hong-Kee
    • Journal of Pharmaceutical Investigation
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    • v.34 no.5
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    • pp.369-377
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    • 2004
  • The objective of this study was to investigate the patterns of protein leaching to an external phase during an ethyl acetate-based, double emulsion microencapsulation process. An aqueous protein solution (lactoglobulin, lysozyme, or ribonuclease; $W_1$) was emulsified in ethyl acetate containing poly-d,l-lactide-co-glycolide 75:25. The $W_1/O$ emulsion was transferred to a 0.5% polyvinyl alcohol solution saturated with ethyl acetate $(W_2)$. After the double emulsion was stirred for 5, 15, 30, or 45 min, additional 0.5% polyvinyl alcohol $(W_3)$ was quickly added into the emulsion. This so-called quenching step helped convert emulsion microdroplets into microspheres. After 2-hr stirring, microspheres were collected and dried. The degree of protein leaching to $W_2$ and/or $W_3$ phase was monitored during the microencapsulation process. In a separate, comparative experiment, the profile of protein leaching to an external phase was investigated during the conventional methylene chloride-based microencapsulation process. When ethyl acetate was used as a dispersed solvent, proteins continued diffusing to the $W_2$ phase, as stirring went on. Therefore, the timing of ethyl acetate quenching played an important role in determining the degree of protein microencapsulation efficiency. For example, when quenching was peformed after 5-min stirring of the primary $W_1/O$ emulsion, the encapsulation efficiencies of lactoglobulin and ribonuclease were $55.1{\pm}4.2\;and\;45.3{\pm}7.6%$, respectively. In contrast, when quenching was carried out in 45 min, their respective encapsulation efficiencies were $39.6{\pm}3.2\;and\;29.9{\pm}11.2%$. By sharp contrast, different results were attained with the methylene-chloride based process: up to 2 hr-stirring of the primary and double emulsions, less than 5% of a protein appeared in $W_2$. Afterwards, it started to partition from $W_1\;to\;W_2/W_3$, and such a tendency was affected by the amount of PLGA75:25 used to make microspheres. Different solvent properties (e.g., water miscibility) and their effect on microsphere hardening were to be held answerable for such marked differences observed with the two microencapsulation processes.