• Title/Summary/Keyword: shake flask

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Biosurfactant Production from Novel Air Isolate NITT6L: Screening, Characterization and Optimization of Media

  • Vanavil, B.;Perumalsamy, M.;Rao, A. Seshagiri
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1229-1243
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    • 2013
  • In this paper, an air isolate (NITT6L) has been screened based on hemolytic activity, emulsification activity, drop collapsing test, and oil displacement test, as well as lipase activity. It was found that strain NITT6L was able to reduce the surface tension of the medium from 61.5 to 39.83 mN/m and could form stable emulsions with tested vegetable oils. Morphological, biochemical, 16S rRNA sequencing analyses, and fatty acid methyl ester analysis using gas chromatography confirmed that the air isolate under study was Pseudomonas aeruginosa. Characterization of the biosurfactant using agar double diffusion assay revealed that the biosurfactant was anionic in nature, and CTAB-methylene blue assay and Molisch test revealed its glycolipid nature. The FT-IR spectrum confirmed that the crude biosurfactant was a rhamnolipid. Using unoptimized medium containing sucrose as the carbon source, the isolate was found to produce 0.3 mg/ml of rhamnolipid in batch cultivation (shake flask) at $37^{\circ}C$ and pH 7. Optimization of the medium components was carried out using design of experiments and the yield of rhamnolipid has been enhanced to 4.6 mg/ml in 72 h of fermentation.

Expression and Secretion of Human Serum Albumin in the Yeast Saccharomyces cerevisae

  • Kang, Hyun-Ah;Jung, Moon-Soo;Hong, Won-Kyoung;Sohn, Jung-Hoon;Choi, Eui-Sung;Rhee, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.42-48
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    • 1998
  • In order to maximize the secretory expression of human serum albumin (HSA) in the yeast Saccharomyces cerevisiae, a series of HSA expression vectors were constructed with a combination of different promoters, 5' untranslated regions (5'UTR), and secretion signal sequences. The expression vector composed of the galactose-inducible promoter GALl0, the natural 5'UTR, and the natural signal sequence of HSA directed the most efficient expression and secretion of HSA among the constructed vectors when introduced into several S. cerevisiae strains. Although the major form of HSA expressed and secreted in the yeast transformants was the mature form of 66 kDa, the truncated form of 45 kDa was also detected both in the cell extract and in the culture supernatant. The level of the intact HSA protein in the culture supernatant reached up to 30 mg/l at 24 h of cultivation in a shake-flask culture but began to decrease afterwards, indicating that the secreted HSA protein was unstable in a prolonged culture of yeast.

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Effect of Galactose and Dextrose on Human Lipocortin I Expression in Recombinant Saccharomyces cerevisiae Carrying Galactose-Regulated Expression System

  • Nam, Soo-Wan;Seo, Dong-Jin;Rhee, Sang-Ki;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.3 no.3
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    • pp.168-173
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    • 1993
  • The expression kinetics of human lipocortin I (LCI), a potential anti-inflammatory agent, was studied in the shake-flask and fermenter cultures of Saccharomyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LCI, and the plasmid stability were investigted under various galactose induction conditions. The expression of LCI was repressed by the presence of a very small amount of dextrose in the culture medium, but it was induced by galactose after dextrose became completely depleted. The optimal ratio of dextrose to galactose for lipocortin I production was found to be 1.0 (10 g/l dextrose and 10 g/l galactose). With optimal D/G ratio of 1.0 and the addition of galactose prior to dextrose depletion, LCI of about 100~130 mg/l was produced. LCI at a concentration of 174 mg/l was porduced in the fed-batch culture, which was nearly a twice as much of that produced in the batch culture. The plasmid stability was very high in all culture cases, and thus was considered to be not an important parameter in the expression of LCI.

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Effect of pH on the elaboration of pullulan and the production of high molecular weight pullulan by Aureobasidium pullulans.

  • Kim, Jeong-Hwa;Zhu, Il-hui;Kim, Mi-Ryeong;Lee, Ji-Hyeon;Kim, Seong-Gu
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.380-383
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    • 2000
  • The effect of on the cell growth, the elaboration of pullulan, the morphology and were the effect of on the molecular weight of pullulan were investigated. A. pullulans showed maximum pullulan production when initial pH 6.5 was 11.98 g/l in shake-flask culture. In batch culture, the maximum pullulan production of 15.16 g/l was obtained at an aeration rate of 0.5 vvm. The mixture of yeast-like form and mycelial form of cells was found at the constant pH 4.5, at which condition, the elaboration of pullulan was high, about 13.31 g/l. However, pullulan with its higher molecular weight (>1,000,000) was produced at the constant pH 6.5.

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A Study on the Antimicrobial Finishing of Artificial Suede by Allylamine Copolymers (Allylamine계 항균제를 이용한 인조스웨드 직물의 항균코팅에 관한 연구)

  • 김윤정;이종우;윤남식
    • Textile Coloration and Finishing
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    • v.12 no.1
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    • pp.61-67
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    • 2000
  • This study was carried out to develope antimicrobial artificial suede by coating with water soluble polyurethane resin and the copolymer of N,N'-dialkyl-N,N'-dialkyl ammonium chloride (DADAAC) and acrylamide as a antimicrobial additve. The copolymer of DADAAC and acrylamide was synthesized by free radical initiation and intra-intermolecular propagation, and the prepared copolymers had sufficient compatibility with water soluble polyurethane resin. The MIC values of the prepared copolymers and antimicrobial characteristics of the artificial suede coated by polyurethane were evaluated. With the increase in the proportion of DADAAC, which is antimicrobially active part in the DADAAC/acrylamide copolymers, the MIC value becomes lower. The MIC value of DADAAC-AA (1 : 1) copolymer is below 30 ppm against S. aureus, and below 90 ppm against K pneumoniae. The artificial suede coated by water soluble polyurethane resin with 1.0% owl concentration of DADAAC/acrylamide copolymer has good antimicrobial fastness as to show colony reduction of above 90% and 80% against S. aureus and K. pneumoniae respectively in the shake flask test after 10 times of washing, and above 95% and 85% after 10 times of dry-cleaning. The elastic recovery of coated suede fabric is not affected up to 1.0% owf concentration of DADAAC-AA copolymer in the polyurethane coating.

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Strain Improvement for Enhanced Production of Streptokinase and Streptodornase in Streptococcus sp.

  • HYUN, HYUNG-HWAN;YOON-BUM LEE;KYUNG-HWA SONG;JI-YOUNG JEON;HYUNE-HWAN LEE
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.101-106
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    • 1997
  • Strain improvement for the enhanced production of streptokinase and streptodornase in Streptococcus sp. ATCC 12449 was performed. Strain UB111, a hyperproductive mutant which was isolated by use of nitrosoguanidine and selection of colonies with large clear zones on DNase test agar plates supplemented with $1{\%}$ glucose and $0.5{\%}$ ammonium chloride, produced about 3 fold more streptokinase and streptodornase than the wild type when tested in shake flask fermentations. The enhanced production of both streptokinase and streptodornase was achieved by cultivating the mutant in a pH-controlled fermentor containing fermentation medium enriched with yeast extract ($2.1{\%}$). Under these conditions, the mutant produced 7300 units/ml of streptokinase and 800 units/ml of streptodornase.

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Xylitol Production from D-Xylose by Candida mogii ATCC 18364 (Candida mogii ATCC 18364를 이용한 D-Xylose로부터 Xylitol 생산)

  • 백승철;권윤중
    • KSBB Journal
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    • v.19 no.3
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    • pp.226-230
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    • 2004
  • Fermentation characteristics of D-xylose into xylitol by Candida mogii ATCC 18364, a potential xylitol producer from rice straw hemicellulose hydrolyzates, were investigated. The influences of the most important operational variables on xylitol production were examined. The best results in xylitol production were obtained in shake-flask fermentations when 3.0 g/L initial cell concentration of 12 hr-old cells grown in D-glucose containing medium were used as inoculum. The oxygen availability is a critical factor in xylose fermentation, therefore, xylose conversion into xylitol was investigated in a 2-L fermenter at different stirring rates. Maximum xylitol production was obtained with an aeration rate of 1 vvm at a stirring rate of 200 rpm.

Breeding of L(+)-Lactic Acid Producing Strain by Low-Energy Ion Implantation

  • Ge, Chun-Mei;Gu, Shao-Bin;Zhou, Xiu-Hong;Yao, Jian-Ming;Pan, Ren-Rui;Yu, Zeng-Liang
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.363-366
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    • 2004
  • In order to obtain an industrial strain with higher L(+)-lactic acid yield, the wild type strain Rhizopus oryzae PW352 was mutated by means of Nitrogen ions implantation (l5 Kev, $7.8 \times 10^{14} - 2.08 \times 10^{15} ions/Cm^2$ and two mutants RE3303 and RF9052 were isolated. After 36 h shake-flask cultivation, the concentration of L(+)-lactic acid reached 131-136 g/l, the conversion rate of glucose was as high as 86%-90% and the productivity was 3.61 g/l.h. It was almost a 75% increase in lactic acid production compared with the wild type strain. Maximum fermentation temperature of RF9052 was increased to $45^{\circ}C$ from original $36^{\circ}C$. At the same time, the preferred range of fermentation temperature of RF9052 was broadened compared with PW352.

Optimization of Medium Composition and Cultivation Parameters for Fructosyltransferase Production by Penicillium aurantiogriseum AUMC 5605

  • Farid, Mohamed Abdel-Fattah Mohamed;Kamel, Zinat;Elsayed, Elsayed Ahmed;El-Deen, Azza Mohamed Noor
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.209-218
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    • 2015
  • Fructooligosaccharides have been mainly produced by microbial fructosyltransferases (FTase) enzymes. The present work focuses on the optimization of medium composition and cultivation parameters affecting FTase produced by Penicillium aurantiogriseum AUMC 5605 in shake flask cultivation. FTase production was optimized in two steps using DeMeo's fractional factorial design. A 1.46-fold increase in FTase production (105.4 U/mL) was achieved using the optimized culture medium consisting of (g/L): sucrose, 600; yeast extract, 10; $K_2HPO_4$, 5; $MgSO_4{\cdot}7H_2O$, 0.5; $(NH_4)_2SO_4$, 1.0 and KCl, 0.5. The obtained results showed that the maximum FTase enzyme activity was produced at initial cultivation pH values ranging from 6.0-6.5, at agitation speed of 200 rpm and using vegetative fungal cells as inoculum. Moreover, results showed that optimization of medium composition and some cultivation parameters resulted in an increase of about 93.7% in the enzyme activity than the nonoptimized cultivation conditions after 96 h of cultivation. Additionally, maximum production and specific production rates recorded 2340 U/L/h and 102 U/L/h/g cells, respectively.

Continuous Production of Citric Acid from Dairy Wastewater Using Immobilized Aspergillus niger ATCC 9142

  • Kim, Se-Kwon;Park, Pyo-Jam;Byun, Hee-Guk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.89-94
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    • 2002
  • The continuous production of citric acid from dairy wastewater was investigated using calcium-alginate immobilized Asrergillus niger ATCC 9142. The citric acid productivity and yield were strongly affected by the culture conditions. The optimal pH, temperature, and dilution rate were 3.0, 30$^{\circ}C$, and 0.025 h$\^$-1/, respectively. Under optimal culture conditions, the maximum productivity, concentration, and yield of citric acid produced by the calcium-alginate immobilized Aspergillus niger were 160 mg L$\^$-1/ h$\^$-1/, 4.5 g/L, and 70.3%, respectively, The culture was continuously perfored for 20 days without any apparent loss in citric acid productivity. Conversely, under the same conditions with a batch shake-flask culture, the maximum productivity, citric acid concentration, and yield were only 63.3 mg L$\^$-1/h$\^$-1/, 4.7 g/L and 51.4%, respectively, Therefore, the results suggest that the bioreactor used in this study could be potentially used for continuous citric acid production from dairy wastewater by applying calcium-alginate immobilized Aspergillus niger.