• Title/Summary/Keyword: Substrate culture

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Induction of Arabidopsis thaliana Chitinase by Ethylene and Elicitor Treatment (에틸렌 및 Elicitor처리에 의한 아기장대풀의 키틴 가수분해 효소 유도)

  • Kyung Hee PAEK;Seok Yoon KWON;Hye Sun CHO;Jin Sam YOU
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.6
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    • pp.357-362
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    • 1994
  • Chitinases and $\beta$-1,3-glucanases are believed to be important in defending plane against pathogens. Here, we investigated the expression of chitinase(s) in Arabidopsis thaliana cell suspension culture system in response to ethephon (2-chloroethyl phosphonic acid) which produces ethylene or a microbial elicitor, a bacterial pectin-degrading enzyme, $\beta$-1, 4-endopolygalactronic acid Iyase (PGA Iyase), treatment. Chitinase activity was measured either by radio chemical assay using $^3$H-labeled regenerated chitin as substrate or western blot analysis using antibody raised against tobacro chitinase(S). With 1 mg/mL of ethephon or 100 m units/mL of elicitor treatment, maximum levels of activity were reached after 48h. We also investigated distribution of chitinase activity in seedlings, leaves, and root of A. thaliana and found that root have the highest chitinase activity.

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Production of casein phosphopeptides using Streptococcus faecalis var. liquefaciens cell immobilization (Streptococcus faecalis var. liquefaciens 전세포 고정화법을 이용한 Casein Phosphopeptides 생산)

  • Lee, Ki-Sung;Shin, Jae-Yoon;Jang, Yi-Hyun;Kweon, Dae-Hyuk;Park, Ki-Moon;Jin, Yong-Su
    • KSBB Journal
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    • v.23 no.1
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    • pp.59-64
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    • 2008
  • Optimum conditions for production of casein phosphopeptides (CPP) from sodium casenate by immobilized cell culture of Streptococcus faecalis var. liquefaciens were investigated. Immobilized cells were made by mixing 60% sodium alginate solution with an equal volume of culture broth at the end of exponential phase and subsequently dropping the mixture into $CaCl_{2}$ solution. Optimum conditions for CPP production by the immobilized cells were the same as those ($50^{\circ}C$, pH 7.0, and 10% substrate concentration) by the crude enzyme solution from the supernatant of culture broth. Optimum loading volume of the immobilized cells into a batch reactor was 30% (w/v). Using a continuous reactor loaded by the immobilized cells under the identified optimal conditions, we were able to produce CPP continuously up to 30 days with a maximum CPP conversion efficiency of 20%.

Overexpression of the spr D Gene Encoding Strptomyces griseus Protease D Stimulates Actinorhodin Production in Streptomyces lividans

  • Choi, Si-Sun;Chi, Won-Jae;Lee, Jae-Hag;Kang, Sang-Soon;Jeong, Byeong-Chul;Hong, Soon-Kwang
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.305-313
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    • 2001
  • The spr D gene encoding Strptomyces griseus protease D(SGPD); a chymotrypsin-like proteae, was cloned from Strptomyces griseus IFO13350 and sequence. Most of the amino-acid sequence deduced from the nucleotide sequence is idential to that Strptomyces griseus IMRU3499 except that one amino acid has been deleted and Trp 369 has been substituted into Cys369 in the SGPD from S. griseus IFO13350 without affecting the protease activity. The spr D gene was overexpressed in Streptomyce liv-idans TK24 as a heterologous host. Various media with different compositions were also used to max-imize the productivity of SGPD inthe heterologous host. The SGPD productivity was best when the transformant S. lividans TK24 was cultivated in R2YE medium. The relative chymotrypsin activity of the culture broth measured with an artificial chromogenic substrate, N-scuccinyl-ala-ala-pro-phe-p-nitroanilide, was 16 units/ml. A high level of SGPD was also produced in YEME and SAAM medial but it was relatively lower that in R2YE medium and negligible amounts of SGPD were produced in GYE, GAE and Benedict media. The growth of S. lividans reacted the maximum level of cell mass at days 3 and 4 of the culture, but SGPD production started in the stationary phase of cell growth and kept increase in till the 10$^{th}$ day of culture in R2YE and YEME medium, but in GYE media the productivity reached maximum level at 8days of cultivation. The introduction of the spr D gene into S. lividans TK24 triggered biosyntheis of the pigmented antibiotic , actinorhodin, which implies some protease may paly a very improtant role in secondary-metabolite formation in sStreptomyces.

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Manufactures of Functional Kimchi using Bifidobacterium Strain Producing Conjugated Linoleic Acid (CLA) as Starter (Conjugated linoleic acid(CLA) 생성균주를 starter로 이용한 기능성 김치의 제조)

  • Min, Sung-Gi;Kim, Jung-Hee;Kim, So-Mi;Shin, Hong-Sig;Hong, Gun-Hwa;Oh, Duk-Gun;Kim, Kyung-Nam
    • Korean Journal of Food Science and Technology
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    • v.35 no.1
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    • pp.111-114
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    • 2003
  • Conjugated linoleic acid (CLA), known to possess various beneficial effects such as anticarcinogenic, antioxidative, and cholesterol-depressing, has been used as a health supplementary food in Japan and the USA. Optimum condition for CLA production without causing changes in quality of kimchi was determined using Bifidobacterium sp., a CLA-producing microorganism, as a starter in culture broth, freeze-dried culture, and encapsulated culture. Results revealed encapsulation was most ideal for maintaining the ability of bacterium to produce CLA during kimchi fermentation. Exogenous linoleic acid (LA) which is a substrate for conversion to CLA was not added to kimchi since LA was already exists in red pepper. Changes in sensory properties of kimchi and production of CLA were measured after inoculation of the encapsulated starter. The optimum inoculation concentration of the encapsulated starter was 0.1% (w/w) for production of CLA without causing changes in kimchi taste.

Separation of Human Breast Cancer and Epithelial Cells by Adhesion Difference in a Microfluidic Channel

  • Kwon, Keon-Woo;Choi, Sung-Sik;Kim, Byung-Kyu;Lee, Se-Na;Lee, Sang-Ho;Park, Min-Cheol;Kim, Pil-Nam;Park, Suk-Ho;Kim, Young-Ho;Park, Jun-Gyul;Suh, Kahp-Y.
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.7 no.3
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    • pp.140-150
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    • 2007
  • A simple, label-free microfluidic cell purification method is presented for separation of cancer cells by exploiting difference in cell adhesion. To maximize the adhesion difference, three types of polymeric nanostructures (50nm pillars, 50nm perpendicular and 50nm parallel lines with respect to the direction of flow) were fabricated using UV-assisted capillary moulding and included inside a polydimethylsiloxane (PDMS) microfluidic channel bonded onto glass substrate. The adhesion force of human breast epithelial cells (MCF10A) and human breast carcinoma (MCF7) was measured independently by injecting each cell line into the microfluidic device followed by culture for a period of time (e.g., one, two, and three hours). Then, the cells bound to the floor of a microfluidic channel were detached by increasing the flow rate of medium in a stepwise fashion. It was found that the adhesion force of MCF10A was always higher than that of MCF cells regardless of culture time and surface nanotopography at all flow rates, resulting in a label-free detection and separation of cancer cells. For the cell types used in our study, the optimum separation was found for 2 hours culture on 50nm parallel line pattern followed by flow-induced detachment at a flow rate of $300{\mu}l/min$.

Biosynthesis of trifolin, a bioactive flavonoid by biotransformation (생물전환으로 생리활성물질인 trifolin의 생합성)

  • Noh, Hye-Ryeong;Kang, Ju-Yeong;Kim, Bong-Gyu
    • Journal of Applied Biological Chemistry
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    • v.64 no.3
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    • pp.309-316
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    • 2021
  • Kaempferol 3-O-galactoside (Trifolin), a member of the flavonol group, has been reported to have anticancer effects against promyelocytic leukemia, histocytic lymphoma, skin melanoma and lung cancer. Trifolin has been extracted and used from several plants, but the extraction process is complicated and the final yield is low. Biotransformation is an alternative tool to produce high value-added chemicals from inexpensive compounds. To synthesis trifolin from naringenin, three genes (PeFLS and OsUGE-PhUGT) were introduced into Escherichia coli, respectively. In order to synthesis trifolin from naringenin, a co-culture fermentation system was established by optimizing the cell concentration, biotransformation temperature and medium, isopropyl-β-D-thiogalactoside (IPTG) concentration, substrate supply concentration, and recombinant protein induction time. The established optimal conditions for trifolin production were a 3:1 ratio of BL-UGTE to BL-FLS, induction of recombinant protein at 25 ℃ for 4 h after addition of 2.0 mM IPTG, biotransformation at 30 ℃, and supply of 300 μM naringenin. Through the optimized co-culture fermentation system, trifolin was biosynthesized up to 67.3 mg/L.

Occurrence of Fusarium wilt and Twospotted Spider Mite under Plastic Mulched and Non-Plastic Mulched Bed in Hydroponic Culture of Strawberry (딸기 수경재배에서 베드 피복유무에 따른 시들음병과 점박이응애 발생 양상)

  • Nam, Myeong Hyeon;Kim, Hyun Sok;Kim, Tae Il;Oh, Sang-Keun
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.257-264
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    • 2018
  • Hydroponic strawberry culture system is increasing annually. Most of strawberry farmers use mulched bed in hydroponic culture and strawberry plants were transplanted in early September. After transplanting, Fusarium wilt caused by Fusarium oxysporum f. sp. fragariae and twospotted spider mite (TSSM), Tetranychus urticae, can increase their occurrence under high temperature condition. Therefore, we conducted for comparison occurrence of Fusarium wilt and TSSM on mulched with green polyethylene film and non-mulched bed. Occurrence of Fusarium wilt on mulched bed was started from early October and more increase than non-mulched bed. Damage rate of TSSM on mulched bed was shown higher than non-mulched bed. Temperature of substrate in mulched bed increased than non-mulched bed, but relative humidity near plants was decreased. As a result, use of non-mulched bed should be effective for reducing of Fusarium wilt and TSSM on strawberry plants.

The Biocompatibility Of Cultured Bone Marrow Cells And Gingival Fibroblasts On The Titanium Surfaces (티타늄 배양에 대한 배양골수와 치은 섬유아세포의 생체적합성)

  • Oh, Choong-Young;Park, Joon-Bong;Kwon, Young-Hyuk;Lee, Man-Sup
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.143-160
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    • 1996
  • The purpose of this study was to evaluate the response in aspect of attachment and growth rate of osteoblasts and growth rate of osteoblasts and human gingival fibroblasts to the commercially pure titanium(CP titanium)and titanium alloy(Ti-6AI-4V) that are used widely as implant materials, and to obtain the basic information to ideal implant materials. In the studly, commercially pure titanium in first test group, titanium alloy(Ti-6AI-4V) in second test group, cobalt-chrome-molybdenum alloy(Co-Cr-Mo alloy) in positive control group, and tissue culture polystyrene plate in negative control group were used. The results of this study were as follows. 1. Bone marrow cells cultured on CP titanium and Ti-6Al-4V showed significantly greater attachment and growth rate(p(0.05) compared to Co-Cr-Mo alloy in each time. 2. There were no significant differences(p>0.05) in attachment and growth rate of bone marrow cells cultured on CP titanium and Ti-6AI-4V or tissue culture plate. 3. Most bone marrow cells cultured on CP titanium, Ti-6Al-4V and tissue culture plate were attached well to each substratum in first 2days, and then, grew at higher growth rate. On the other hand, some cells cultured on Co-Cr-Mo alloy failed to attach in first 2 days, and then, attached cells grew at lower growth rate than other groups. 4. Attachment and growth rates of gingival fibroblasts cultured on CP titanium and Ti-6Al-4V showed no significant differences(p>0.05) compared to Co-Cr-Mo alloy in 2 days, but significantly greater increase(p<0.05) in 5 and 9 days. 5. There were no significantly differences(p>0.05) between growth rates on gingival fibroblasts cultured on CP titanium, Ti-6Al-4V and tissue culture plate in 2 and 5days, but a significant lower growth rate(p<0.05) on CP titanium and Ti-6Al-4V versus tissue culture plate. 6. Some gingival fibroblasts cultured on all specimen groups failed to attach, but attached cells grew well, especially on CP titanium, Ti-GAl-4V and tissue culture plate. 7. There were no significant differences(P>0.05) between growth rates of both bone marrow cells and gingival fibroblasts cultured on CP titanium and Ti-6AI-4V. As a result of this study, both commercially pure titanium and Ti-6AI-4V showed excellent biocompatibility and there was no significant difference in the cellular response to the both metals. Bone marrow cells cultured on each substratum showed significantly greater growth rate and responded sensitively to cytotoxic effects of metal surfaces compared to gingival fibroblasts. Considering cell response to the substrate, it was likely that the composition itself of titanium metals have no significant effect on the biocompatibility. Further study need to be done to evaluate the influence of surface characteristics on cellular responses.

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Ethanol Production from Tapioca Hydrolysate by Batch and Continuous Cell Retention Cultures (회분 및 연속세포유지 배양에 의한 타피오카당화액으로부터의 에탄올생산)

  • 이용석;이우기
    • KSBB Journal
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    • v.10 no.5
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    • pp.598-603
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    • 1995
  • Batch and continuous cell retention cultures were carried out using tapioca hydrolysate. In batch culture, reducing sugar of about 180g/$\ell$ was almost consumed in about 36 hours, and the concentration of ethanol produced was about 84g/$\ell$ making the ethanol yield 0.48 g-ethanol/g-(reducing sugar). The final yeast concentration was 8.5${\times}$107 cells/ml(about 2.1g/$\ell$). In a total cell retention culture operated with a dilution rate of 0.18h-1, the yeast concentration, the residual reducing sugar concentration, the ethanol concentration, and the volumetric ethanol productivity were about 40g/$\ell$, about 15g/$\ell$, 81.4g/$\ell$, and 14.7g/$\ell$-h, respectively. In another cell retention culture operated with a dilution rate and a bleed ratio of 0.2h-1 and 0.14, respectively, the yeast concentration increased to 22g/$\ell$ and the ethanol concentration oscillated around 68g/$\ell$. The volumetric ethanol productivity was about 13.6g/$\ell$-h and the residual reducing sugar concentration about 12g/$\ell$ containing glucose of about 4.5g/$\ell$. According to the results of batch fermentation using the solid residue from hydrolysate filtration as the substrate, it seemed to have a certain value. Thus, development of an effective reactor system to produce ethanol from this solid residue is in need.

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Investigation on Favorable Substrate Formulation for Bag Cultivation of Grifola frondosa (잎새버섯 봉지재배에 적합한 배지조성 연구)

  • Kim, Jeong-Han;Choi, Jong-In;Chi, Jeong-Hyun;Won, Sun-Yee;Seo, Geon-Sik;Ju, Young-Cheoul
    • The Korean Journal of Mycology
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    • v.36 no.1
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    • pp.26-30
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    • 2008
  • This study was carried out to develop artificial culture method of Grifola frondosa in polypropylene bag. To find out the favorable substrate formulation of G. frondosa, physicochemical conditions were investigated. The T2 formulation (55 : 25 : 12 : 8) mixing ratio of oak sawdust, oak chip, dried bean-curd refuse and wheat bran showed the shortest time to complete the crop cycle and the highest yield (weight of fresh mushrooms harvested at maturity). Those physicochemical conditions were pH 4.3, 2.4% crude fat contents, 54.1% total carbon, 1.42% total nitrogen, 38.1 C/N ratio, 75.5% porosity and $0.21\;g/cm^3$ bulk density. Among the physicochemical factors, pH, crude fat and total nitrogen may affect mushroom yield. Therefore, development of favorable substrate would be benefit to increase production efficiency of G. frondosa, mushrooms and be commercial potential.