• Title/Summary/Keyword: dry cell weight

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Isolation and Characterization of Purple Non-Sulfur Bacteria, Afifella marina, Producing Large Amount of Carotenoids from Mangrove Microhabitats

  • Soon, Tan Kar;Al-Azad, Sujjat;Ransangan, Julian
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1034-1043
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    • 2014
  • This study determined the effect of light intensity and photoperiod on the dry cell weight and total amount of carotenoids in four isolates of purple non-sulfur bacteria obtained from shaded and exposed microhabitats of a mangrove ecosystem in Kota Kinabalu, Sabah, Malaysia. The initial isolation of the bacteria was carried out using synthetic 112 medium under anaerobic conditions (2.5 klx) at $30{\pm}2^{\circ}C$. On the basis of colony appearance, cell morphology, gram staining, motility test, and 16S rRNA gene sequencing analyses, all four bacteria were identified as Afifella marina. One of the bacterial isolates, designated as Af. marina strain ME, which was extracted from an exposed mud habitat within the mangrove ecosystem, showed the highest yield in dry cell weight ($4.32{\pm}0.03g/l$) as well as total carotenoids ($0.783{\pm}0.002mg/g$ dry cell weight). These values were significantly higher than those for dry cell weight ($3.77{\pm}0.02g/l$) and total carotenoid content ($0.706{\pm}0.008mg/g$) produced by the isolates from shaded habitats. Further analysis of the effect of 10 levels of light intensity on the growth characteristics of Af. marina strain ME showed that the optimum production of dry cell weight and total carotenoids was achieved at different light intensities and incubation periods. The bacterium produced the highest dry cell weight of 4.98 g/l at 3 klx in 72 h incubation, but the carotenoid production of 0.783 mg/g was achieved at 2.5 klx in 48 h incubation. Subsequent analysis of the effect of photoperiod on the production of dry cell weight and total carotenoids at optimum light intensities (3 and 2.5 klx, respectively) revealed that 18 and 24 h were the optimum photoperiods for the production of dry cell weight and total carotenoids, respectively. The unique growth characteristics of the Af. marina strain ME can be exploited for biotechnology applications.

Effect of Suspension Culture Conditions on Cell Activity of Wild Viola(Viola partrinii DC.) Callus (야생 흰 제비꽃(viola patrinii DC.)callus의 현탁배양 방법이 세포 활성에 미치는 영향)

  • Kim, Du-Hyun;Chung, Yong-Mo;Chung, Chung-Han;Yeeh, Yeehn;Kwon, Oh-Chang
    • Journal of Life Science
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    • v.6 no.2
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    • pp.94-103
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    • 1996
  • To understand effect of inoculum size, cell density, sucrose concentration and concentrations of MS basal on suspension culture and protoplast isolation of wild viola(Viola patrinii DC.) callus from petiole segments this experiment was conducted. In the lot of 30 mesh inoculum size, two observations were; One was that a considerable increase in the fresh and dry weight of callus was determined. Another was that the callus mass was relatively compact compared with others. A recommendable cell density was 0.4g for 20ml culture medium and the higher sucrose concentration, the higher fresh and dry weight were obtained. The dilution of MS basal salt was differently affected on fresh and dry weight; the highest fresh weight was found in 1x MS salt, while the higest dry weight was in 1/3x dilution.The addition of casein hydrolysate(3g/L) was more effective to increase of both fresh and dry weight. THe contents of protein was great in the inoculum lots with larger inoculum sizeand higher concentration of MS basal salts contenting 3g/L of casein hydrolysate and higher sucrose compared with others. The greatest protoplasts were isolated from the lot of 10 mesh size treated with 1%pectinase SE-150 and 2% cellulase YC. In general, for optimal protoplast isolation the followingconditions were recommended; 1) Use of smaller cell size cultured for 2-5 weeks, and 2) more than 5 hours incubation using the combined mixture of the enzymes with proper concentrations.

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Production of Gomisin J from Cell Suspension Cultures of Schisandra chinensis Baillon (오미자의 현탁배양세포로부터 Gomisin J의 생산)

  • Hwang, Sung-Jin;Pyo, Byoung-Sik;Lee, Hak-Ju;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.442-447
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    • 2004
  • Cell growth and gomisin J production by suspension cultures of Schisandra chinensis Baillon were investigated under various culture media, initial sucrose concentrations, shaking speeds, and inoculum sizes. Callus was induced from in vitro cultivated leaf segments on MS medium supplemented with $1\;mg/{\ell}$ NAA. The maximum dry cell weight of 2.23 g was obtained at inoculum size of 0.5 g fresh cell weight and in MB5 medium supplemented with $1\;mg/{\ell}$ NAA, 3% sucrose after 8 weeks. The production of gomisin J in suspension cell cultures was maximized in WPM medium containing 5% sucrose. The shaking speed for maintaining maximal cell dry weight was 100 rpm while the best shaking speed for gomisin J accumulation was 140 rpm.

Estimation of Cell Concentration by Light Transmitter During the Culture of Methylotrophic Yeast Pichia pastoris

  • Choi, Du-Bok;Park, Enoch Y.S.;Lee, Yong-Bo;Na, Young-Hee;Lim, Chae-Kyu
    • Mycobiology
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    • v.31 no.4
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    • pp.226-228
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    • 2003
  • The multiple correlation coefficient between the values determined by dry weight and those determined by fluorometer was observed with r=0.96 and the standard error of calibration was 0.034. Using the best calibration data, in order to reconfirm the reliability of the fluorometer results in comparison with those obtained by dry weight on the cell concentration, fedbatch cultures were carried out. The results obtained by fluorometer measurements were in good agreement with those obtained by dry weight. The on-line monitoring of cell concentration by the fermentor system linked to a computer equipped with fluorometer was successfully carried out.

Heterologous Expression of Human Ferritin H-chain and L-chain Genes in Saccharomyces cerevisiae (재조합 효모를 이용한 사람 H-Chain 교 L-Chain Ferritin의 생산)

  • 서향임;전은순;정윤조;김경숙
    • KSBB Journal
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    • v.17 no.2
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    • pp.162-168
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    • 2002
  • Human ferritin H- and L-chain genes(hfH and hfL) were cloned into the yeast shuttle vector YEp352 with various promoters, and the vectors constructed were used to transform Saccharomyces cerevisiae 2805. Three different promoters fused to hfH and hfL were used: galactokinase 1 (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase(GPD) promoter and alcohol dehydrogenase 1(ADH1 ) promoter. SDS-polyacrylamide gel electrophoresis and Western blotting analyses displayed expression of the introduced hfH and hfL. In the production of both ferritin H and L subunits GAL1 promoter was more effective than GPD promoter or ADH1 promoter. Ferritin H and L subunits produced in S. cerevisiae were spontaneously assembled into its holoproteins as proven on native polyacrylamide gels. Both recombinant H and L-chain ferritins were catalytically active in forming iron core. When the cells were cultured in the medium containing 10 mM ferric citrate, the cell-associated concentration of iron was 174.9 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L and 148.8 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L but was 49.4 $\mu\textrm{g}$ Per gram(dry cell weight) in the wild type, indicating that the iron contents of yeast is improved by heterologous expression of human ferritin H-chain or L-chain genes.

PHB Accumulation Stimulated by Ammonium Ions in Potassium-limited Cultures of Methylobacterium organophilum

  • Kim, Seon-Won;Kim, Pil;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.301-304
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    • 1998
  • Methylobacterium organophilum can use nitrogen in the form of ammonium ions ($($NH_4$)_2SO_4\;and\;NH_4Cl) and from nonammonium sources such as glycine, alanine, peptone, and yeast extract. When potassium was limited, significantly more PHB was produced when the ammonium ion was the nitrogen source rather than a nonammonium form. With ammonium, the amount of PHB produced was 0.50-0.53 g PHB/l or $52.0~53.2\%$ of the dry cell weight. If nitrogen was from a nonammonium source, the respective values were 0.04~0.06 g PHB/1 or $8.1~11.3\%$ of dry cell weight. When ammonium sulfate was the sole source of nitrogen under potassium-limited conditions, cell growth and PHB accumulation increased as the pH increased from 6.0 to 7.5. Cell growth and PHB amount at pH 7.5 were 2.50 g dry cell weight/1 and 1.40 g PHB/1, respectively.

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Effect of Various Carbon Sources on the Production and Stabilization of hGM-CSF in Transgenic Plant Suspension Culture (형질전환된 식물세포에서 hGM-CSF 생산과 안정성에 대한 다양한 탄소원의 효과)

  • Lee Jae-Hwa
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.313-319
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    • 2005
  • The effects of various carbon sources on the secretion of hGM-CSF, total protein and protease into the medium were investigated in transgenic tobacco cells. The dry cell weight (11.2 g/L) and wet cell weight (310.8 g/L) were highest at 30 g/L glucose after 5-day culture but, the dry cell weight (13.4 g/L) and wet cell weight (480 g/L) were highest at 30 g/L sucrose after 10-day culture. The total protein (110.3 mg/L), protease activity (3950 U/L) and total secreted hGM-CSF (56 mg/L) were highest at 30 g/L sucrose after 10-day culture. Stabilization of the total secreted protein and hGM-CSF in various carbon source concentrations was determined. Total secreted protein was most stabilized in the medium containing sucrose. However, the loss of the total protein was increased with the concentrations of high level in medium containing sorbitol, mannitol, fructose, and glucose. hGM-CSF was more stabilized in the medium containing sucrose than in the medium containing sorbitol, mannitol, fructose, glucose.

Studies on the Ploidy of Saccharomyces cerevisiae (Saccharomyces cerevisiae의 배수성에 관한 연구)

  • 조상호;심상국;정동효
    • Microbiology and Biotechnology Letters
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    • v.14 no.4
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    • pp.299-304
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    • 1986
  • The cell volume, cell surface, cell concentration, dry cell weight, frequence of respiratory deficient mutation, resistance against ultraviolet irradiation, fermentation power, DNA contents of haploid diploid, triploid and tetraploid of Saccharomyces cerevisiae strain were investigated. Respiratory deficient mutants by spontaneous mutation were absolved more frequently in the haploid than in the diploid, triploid and tetraploid. And cell volume, cell surface, cell concentration, dry cell weight, resistance against ultraviolet irradiation, fermentation power, and DNA contents were significantly increased as the ploidy increased.

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Effect of Aeration-Agitation on Coenzyme Q10 Production Using Rhodobacter sphaeroides

  • Jeong, Soo-Kyoung;Kim, Joong-Kyun
    • Fisheries and Aquatic Sciences
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    • v.11 no.4
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    • pp.224-228
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    • 2008
  • With the aim of increasing the $CoQ_{10}$ production in mass culture, the effect of aeration-agitation on the $CoQ_{10}$ production using Rhodobactor sphaeroides was investigated in a l-L bioreactor. The maximum $CoQ_{10}$ production was 1.69 mg/g of dry cell weight under conditions of 50 Lux, $30^{\circ}C$, 300 rpm, and 5-vvm aeration. The $CoQ_{10}$ production was improved to produce 2.91 mg/g of dry cell weight under reduced conditions of agitation speed (200 rpm) and aeration rate (0.2 vvm). When R. sphaeroides was cultivated under more reduced DO levels during the exponential phase of the cell, the $CoQ_{10}$ production yield of 3.88-mg/g dry cell weight was the maximum obtained. Therefore, poorer conditions of aeration-agitation resulted in higher production of $CoQ_{10}$, and thus DO content was a crucial factor in the production of $CoQ_{10}$. Accordingly, it was necessary to control the DO concentration in order to enhance the $CoQ_{10}$ biosynthesis within a large-scale production.

Study of Methylglyoxal and Phosphorus Stress on Algae (조류의 Methylglyoxal과 인 Stress 연구)

  • 이기태
    • Environmental Analysis Health and Toxicology
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    • v.13 no.3_4
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    • pp.133-142
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    • 1998
  • Effects of phosphorous (P) and methylglyoxal (MG) on the cell number, dry weight, chlorophyll content, photosynthetic and respiratory rate, phosphate uptake and protein content of green algae (Scenedesrnus obliquus) were studied. The algal cell number from the medium treated with 0.5-1.0 mM of MG at 1/2 P or 1/4 P concentration was significantly lower than those of algae treated :with full strength of phosphrous in medium. The inhibitory effect of MG on algal cell division was enhenced at low concentration of phosphorous in medium. At the beginning of logrithmic phase of algal growth, the mean dry weight of algae from the medium without MG-treatment in 1/2 P media was significantly higher than that of algae treated with MG. After logrithmic phase of growth cycle, the mean dry weight of algae from the medium with 1.0 mM of MG-treatment in 1/4 P media was significantly lower than that of algae treated with or without MG. At logrithmic phase of algal growth, there were significant differences in the chlorophyll content among all groups of tested algae with various concentrations of P and MG. At 15 days after inoculation, the mean chlorophyll content per algal cell from the media without MG-treatment in 1/2P was significantly higher than that of other cells from MG-treated media. The adverse effect of MG at concentration of 0.5-1.0mM in 1/2 and 1/4 P media on photosynthetic rate was observed. The mean photosynthetic rate of algal cell without P and MG treatment at 15 days after inoculation was significantly higher than that of MGtreated algae. After logarithmic phase, the algal cell treated with 0.5mM of MG with full strength of phosphorous showed significantly high respiratory rate than that of other cell groups. There were significant differences in mean phosphate uptake rate among all groups of Scenedesmus obliquus at logarithmic phase. At 12 days after inoculation, phosphate uptake rate per each algal cell from the basic media without MG and P treatment was rapidly reduced which shows early introduction to stationary phase.

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