• Title/Summary/Keyword: Substrate culture

Search Result 656, Processing Time 0.031 seconds

Simple Formation of Poly(sodium 4-styrenesulfonate) Pattern on the Hydrophobic Substrate for the Control of Cell Adhesion via a Selective Ion Irradiation

  • Kim, Soo-Jung;Hwang, In-Tae;Jung, Jin-Mook;Jung, Chan-Hee
    • Journal of Radiation Industry
    • /
    • v.7 no.2_3
    • /
    • pp.149-154
    • /
    • 2013
  • In this study, the simple preparation of poly(sodium 4-styrenesulfonate) (PSS)-patterned substrate via a selective ion irradiation was investigated to manipulate cell adhesion. PSS thin films spin-coated onto the hydrophobic polystyrene (PS) was patterned through masked 150 keV proton irradiation followed by developing with deionized water. The characteristics of the resulting PSS-patterned surfaces were investigated by using microscope, surface profiler, FT-IR, XPS, and contact angle analyzer. These analytical results revealed that the resolved $100{\mu}m$ PSS patterns were formed on the hydrophobic PS surface above the fluence of $1{\times}10^{15}ions\;cm^{-2}$ and the chemical structure, composition, and wettability of the PSS patterns were dependant on a fluence. Moreover, the results of the in-vitro cell culture and proliferation assay exhibited that H1299 cells preferentially adhered and proliferated onto the more hydrophilic PSS part of the PSS-patterned PS and the well-aligned cell patterns was formed on the PSS-patterned PS particularly at the fluence of $1{\times}10^{15}ions\;cm^{-2}$.

Characterization of Low-Temperature Enzymatic Reactions through Heterologous Expression and Functional Analysis of Two Beta-Glucosidases from the Termite Symbiotic Bacterium Elizabethkingia miricola Strain BM10

  • Dongmin LEE;Tae-Jong KIM
    • Journal of the Korean Wood Science and Technology
    • /
    • v.51 no.4
    • /
    • pp.270-282
    • /
    • 2023
  • Lower termites need symbiotic microbes for cellulose digestion. Elizabethkingia miricola strain BM10 has been proposed as a symbiotic microbe that assists in low-temperature digestion and metabolism of Reticulitermes speratus KMT1, a termite on Bukhan Mountain, Seoul, Korea. In E. miricola strain BM10, β-glucosidase genes expressed at 10℃ were identified, and the psychrophilic enzymatic characteristic was confirmed by heterogeneously expressed proteins. Crude β-glucosidase in the culture broth of E. miricola strain BM10 showed specific enzymatic properties, and its substrate affinity was 4.69 times higher than that of Cellic CTec2. Among the genes proposed as β-glucosidase, two genes, bglB_1 and bglA_2, whose gene expression was more than doubled at 10℃ than at 30℃, were identified. They were heterogeneously expressed in Escherichia coli and identified as psychrophilic enzymes with an optimal reaction temperature of about 20℃-25℃. In this study, E. miricola strain BM10, a symbiotic bacterium of lower termites, produced psychrophilic β-glucosidases that contribute to the spread of the low-temperature habitat of a lower termite, R. speratus KMT1.

Biodegradation Kinetics of Phenol and pcresol by Micrococcus sp. M1 (Micrococcus sp. M1에 의한 Phenol과 p-Creso의 생분해 Kinetics)

  • Son, Hong-Joo;Jang, Woong-Seok;Lee, Geon;Lee, Sang-Joon
    • Journal of Environmental Science International
    • /
    • v.6 no.2
    • /
    • pp.153-163
    • /
    • 1997
  • In order to fad the most fitted biodegradation model, biodegradation kinetics model to the initial phenol and p-cresot concentrations were investigated and had been fitted by the linear regression. Bacteria capable of degrading p-cresol were isolated from soil by enrichment culture technique. Among them, strain Ml capable of degradillg p.rcresol has also degraded phenal and was identified as the genus Micrococcus from the results from of taxonomical studies. The optimal tonditlons for the biodegradation of phenal and p-cresol by Micrococcus sp. Ml were $NH_4NO_3$ 0.05%, pH 7.0, 3$0^{\circ}C$, respectively, and medium volume 100m1/250m1 shaking flask. iwicrococcus sp. Ml was able to grow on phenal concentration up to 14mM and p-cresol concelltration up to 0.8mM. With increasing substrate concentraction, the lag period increased, but the maximum specific growth rates decreased. The yield coefficient decreased with increasing substrate concentation. The biodegradation kinetics of phenol and p-cresol were best described by Monod with growth model for every experimented concentration. In cultivation of mixed substrate, p-cresol was degraded first and phenol was second. This result implies that p-cresol and phenol was not degraded simultaneously.

  • PDF

Surface Characteristics and Biocompatibility of Titanium Coated with Dentin-derived Hydroxyapatite

  • Kim, Hae-Jin;Son, Mee-Kyung;Lee, Kyung-Ku;Lee, Bo-Ah;Kim, Young-Joon
    • International Journal of Oral Biology
    • /
    • v.37 no.1
    • /
    • pp.9-16
    • /
    • 2012
  • The aim of this study was to evaluate surface characteristics and biological properties of the dentin -derived hydroxyapatite (HA) coating on titanium substrate. Dentinderived HA was obtained from extracted human teeth using a calcination method at $850^{\circ}C$. The commercially pure titanium (cp-Ti, ASTM Grade II) was used as a metallic substrate and a radio frequency magnetron sputtering method was employed as a coating method. Scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX) were utilized to investigate the coating aspects and composition. Atomic forced microscopy (AFM) and a surface profiler were used to assess the surface morphology and roughness. Corrosion tests were performed in phosphate-buffered saline at a $36.5{\pm}1^{\circ}C$ in order to determine the corrosion behavior of the uncoated and coated specimens. The biocompatibility of dentin-derived HA coated specimens with fetal rat calvarial cells and human gingival fibroblasts was assessed by SEM and cell proliferation analysis. The results showed that the dentin-derived HA coatings appeared to cover thinly and homogeneously the surfaces without changing of the titanium substrate. The EDX analysis of this the coating surface indicated the presence of Ca and P elements. The mean surface roughness of cp-Ti and dentin-derived coating specimens was $0.27{\mu}m$ and, $1.7{\mu}m$, respectively. Corrosion tests indicated a stable passive film of the dentin-derived HA coating specimens. SEM observations of fetal rat calvarial cells and human fibroblast cells on coated surfaces showed that the cells proliferated and developed a network of dense interconnections. The cells on all specimens proliferated actively within the culture period, showing good cell viability. At day 1 and 3, dentin-derived coating specimens showed 89% and 93% cell viability, respectively, when normalized to cp-Ti specimens. These results suggest that dentin-derived HA coating using the RF magnetron sputtering method has good surface characteristics and biocompatibility.

Visible and Fast Assay System for Tobacco Transformant Introduced with Adenosine Deaminase Marker Gene (Adenosine Deaminase 표지유전자로 형질전환된 연초의 신속한 Assay 방법)

  • 양덕춘;김용환;임학태;방극수;배창휴
    • Korean Journal of Plant Tissue Culture
    • /
    • v.28 no.3
    • /
    • pp.165-171
    • /
    • 2001
  • New visible and fast assay system have been developed for tobacco transformant introduced with adenosine deaminase (ADA) marker gene, which converts cytotoxic adenosine analogues to non-toxic inosine analogues and ammonia. Ammonia was changed to blue color in the solution of phenol-nitoprusside and alkaline-hypochlorite. It was possible to detect activity of ADA visibly on the holes of 96 well plate using tiny explant of transgenic tobacco leaves within 1 hour incubation time. As substrates of ADA enzyme from transgenic plant on the plate, a number of adenosine analogues such as 9-D-arabinofuranosyl adenine, cordycepin, 2'-deoxyadenosine, adenosine and xylofuranosyl adenine were possible for detection of ADA activity. Optimal condition of substrate for ADA enzyme was each 10 mM and pH 7.5 in adenosine solution. Especially, transgenic plant did not convert adenosine to inosine and ammonia in the presence of ADA inhibitor deoxycoformycin, which means that ammonia produced from transgenic plant is due to expression of ADA gene. Now, we show that this detection system can be easily, sensitively, fast and cheaply as well as visibly assayed in vitro as GUS gene system with very small size of transformant explant.

  • PDF

Usage of Enzyme Substrate to Protect the Activities of Cellulase, Protease and α-Amylase in Simulations of Monogastric Animal and Avian Sequential Total Tract Digestion

  • Wang, H.T.;Hsu, J.T.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.19 no.8
    • /
    • pp.1164-1173
    • /
    • 2006
  • Cellulase from Aspergillus niger, (${\alpha}$-amylase from Bacillus sp. and protease from Bacillus globigii were used as enzyme sources in this study to examine how their respective substrates protect them in two kinds of simulated gastrointestinal tract digesting processes. Avian total digest tract simulation test showed that filter paper, Avicel and cellulose resulted in 7.7, 6.4 and 7.4 times more activity than of unprotected cellulose, respectively. Protease with addition of casein, gelatin or soybean protein showed no significant protection response. Starch protected amylase to be 2.5 times activity of the unprotected one. Monogastric animal total tract digestion simulation test showed that filter paper, Avicel and cellulose resulted in 5.9, 9.0 and 8.8 times activity of unprotected cellulase, respectively. Casein, gelatin and soybean protein resulted in 1.2, 1.3 and 2.0 times activity of unprotected protease, respectively. Starch did not protect amylase activity in monogastric animal total tract simulation. Protection of mixed enzymes by substrates in two animal total tract simulation tests showed that filter paper in combination with soybean protein resulted in 1.5 times activity of unprotected cellulose, but all substrates tested showed no significant protection effect to protease. Soybean protein and starch added at the same time protected the amylase activity to be two times of the unprotected one. Test of non-purified substrate protection in two animal total digest tract simulation showed that cellulase activity increased as BSA (bovine serum albumin) concentration increased, with the highest activity to be 1.3 times of unprotected enzyme. However, BSA showed no significant protection effect to protease. Amylase activity increased to 1.5 times as BSA added more than 1.5% (w/v). Cellulase activity increased to 1.5 times as soybean hull was added higher than 1.5%. Amylase had a significant protection response only when soybean hull added up to 2%. Protease activity was not protected by soybean hull to any significant extent.

Studies on the Fermentative Utilization of Cellulosic Wastes (Part 7) Culture of Cellulolytic Bacteria from Miscanthus (폐섬유자원의 발효공학적 이용에 관한 연구 (제7보) 억새풀을 기쇄로 한 섬유소 자화세균의 배양)

  • 성낙기;윤한대;심기환;이천수
    • Microbiology and Biotechnology Letters
    • /
    • v.5 no.3
    • /
    • pp.127-131
    • /
    • 1977
  • Various kinds of native herbage grasses like-Miscanthus sinensis, Arundinella hirta, Cymbopogen geirngii, Themeda japonica etc. are widely distributed in every Korean mountain. So we investigated the availability of native grass, Miscanthus sinensis as a substrate for the production of S. C. P. The results were obtained as follows. 1) At the alkali treatment, NaOH was the most effective with the exclusion of lignin, and pretreated Miscanthus with NaOH appeared to be a good substrate for the microbial growth. 2) When Miscanthus was treated with one to 10% NaOH, the microbial growth increased in proportion to the increased alkali concentration. Beyond 4% NaOH, a slight increase was observed. 3) Phosphoric acid, as a neutralizer, was the most effective in cell production after alkali treatment. 4) On the effect of incubation time, the productivity was best found at 60 hours, and the cell weight was 9.23 mg per 1 ml, and the microbial digestibility of substrate was 75.2%.

  • PDF

Characteristics and Antimicrobial Resistance Patterns of Staphylococcus aureus Isolated from Horse (국내 말에서 분리된 Staphylococcus aureus의 특성 및 약제 내성 양상)

  • Choi, Seong-Kyoon;Cho, Gil-Jae
    • Journal of Life Science
    • /
    • v.18 no.1
    • /
    • pp.69-74
    • /
    • 2008
  • This study carried out to investigate the antimicrobial resistance and biochemical characteristics of Staphylococcus aureus (S. aureus) isolated from reproductive and respiratory tract in Thoroughbred horse. The specimens were collected from equine vaginal mucosa and upper respiratory tract from March to December 2006 using a culture swab in Korea. S. aureus suspected colonies on blood agar plates were selected and identified as standard biochemical tests and PCR (Applied Biosystems, USA). Antimicrobial resistance test of S. aureus isolates was performed with 30 antimicrobial agents (BBL, USA) by using the agar disk diffusion method. S. aureus isolates were isolated 58 (39.2%) strains of 148 samples: wound 64.7% (11/17), genital discharge 37.0% (37/100) and nasal discharge 32.2% (10/31). Almost isolates showed high resistance to spectinomycin, sulfonamides, erythromycin, tetracyelin, ciprofloxacin and penicillin. These results may provide the basic information to establish strategies for treatment and prevention of reproductive and respiratory disease in Thoroughbred horses in Korea.

Cultural characteristics of Bacillus velezensis HKB-1 in the water extract of the composted spent mushroom substrate of Lentinula edodes and biological control of Phytophthora blight disease of pepper (표고버섯 수확후배지 퇴비 추출물에서 Bacillus velezensis HKB-1의 배양적 특징 및 고추역병의 생물학적 방제)

  • Kim, Ja-Yoon;Seo, Hyun-Ji;Kang, Dae-Sun;Kang, Hee-Wan
    • Journal of Mushroom
    • /
    • v.19 no.4
    • /
    • pp.272-278
    • /
    • 2021
  • Bacillus velezensis HKB-1 was isolated from the composted spent mushroom substrate of Lentinula edodes (LeCSMS) and inhibited mycelial growth of phytopathogenic fungal species, Phythhopthora capsici, Collectotrichum coccodes and Fusarium oxysporium by more than 70%. B. velezensis HKB-1 showed bacterial growth rate 10 to 100 times higher than that of other commercial bacterial media in water extract of LeCSMS supplemented with 1% molasses. The LeCSMS medium was effective in promoting the growth of pepper and controlling Phytophthora blight disease of pepper.

Comparison of the Influence of Carbon Substrates on the Fibrolytic Activities of Neocallimastix sp. NLRI-3 (탄소원의 종류가 반추위 혐기 곰팡이 Neocallimastix sp. NLRI-3의 섬유소 분해효소 활력에 미치는 영향 비교)

  • 손호진;송재용;최낙진;하종규;장종수
    • Journal of Animal Science and Technology
    • /
    • v.48 no.3
    • /
    • pp.415-424
    • /
    • 2006
  • The purpose of this study was to investigate the fungal growth and enzyme production under different carbohydrate substrate conditions. The anaerobic fungus Neocallimastix sp. NLRI-3 isolated from the rumen of Korean native goat was incubated with different carbohydrate media containing 0.2% of glucose, starch, rice straw, filter paper, carboxymethyl cellulose(CMC), Sigmacell cellulose, xylan or xylose, respectively. The culture head gas production was the highest in the culture of filter paper medium, and the lowest in the culture of CMC medium at 96h incubation (P<0.05). The fungal zoospore production reached peak at 72h incubation, and its number was the highest in rice straw medium among the treatments (P<0.05). At 96h incubation, carboxymethyl cellulase(CMCase) activity was the highest in the culture of filter paper medium and the lowest in the culture of starch medium (P<0.05). While xylanase activity was the highest in the culture of rice straw medium and the lowest in the culture of xylose medium(P<0.05) at 72h incubation. There were no differences in culture supernatant protein expression among the treatments. However, the patterns of enzyme expression were different among the treatments with zymogram analysis. Six CMCases and 4 xylanase were detected from the results of zymogram analysis. Therefore the present study indicating that the fungal enzyme expression could be stimulated with insoluble substrates in the culture medium.