• Title/Summary/Keyword: Batch culture

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Degradation of Salicylic Acid by Free and Immobilized Cells of Pseudomonas sp. Strain NGK1

  • Patil, Neelakanteshwar-K.;Sharanagouda, U.;Niazi, Javed-H.;Kim, Chi-Kyung;Karegoudar, Timmanagouda-B.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.29-34
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    • 2003
  • A Pseudomonas sp. strain NGK1 (NCIM 5120) capable of utilizing salicylate was immobilized in alginate and polyurethane foam (PUF). The degradation rate of salicylate by freely suspended cells was compared with the degradation rate by immobilized cells. In an initial 20 and 40 mM salicylate, free cells ($2{\times}10^{11}\;cfu\;ml^{-1}$) degraded to 16 and 14 mM, alginate-entrapped cells degraded to 18 and 26 mM, and PUF-entrapped cells degraded to 20 and 32 mM salicylate, respectively, in batch cultures. The alginate-and PUF-entrapped cells were used in repeated batch and continuous culture systems. The efficiency of both the immobilized systems f3r the degradation of salicylate was compared. It has been observed that the PUF-entrapped cells could be reused for more than 20 cycles whereas alginate-entrapped cells could be reused for a maximum of only 12 cycles, after which a decrease in degradation rat was observed with the initial 20 and 40 mM salicylate. The continuous degradation of sallcylate by freely suspended cells showed a negligible degradation rate of salicylate when compared with immobilized cells. With the immobilized cells in both alginate and polyurethane foam, the degradation rate increased with an increase in the dilution rate up to $2\;h^{-1}$ for 20 mM, and $1.5\;h^{-1}$ for 40 mM salicylate. The results revealed that PUF-entrapped cells were more efficient for the degradation of salicylate than alginate-entrapped cells and freely suspended cells.

Production of Single-Cell Protein from Methanol (Part 2). Batch Fermentation and Continuous Fermentation by Methylomonas methanolica (Methanol을 이용한 단세포단백질 생산에 관한 연구 (제 2 보), Methylomons methanolica YUFE 101의 회분발효와 연속발효)

  • 유주현;정건섭;변유량
    • Microbiology and Biotechnology Letters
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    • v.7 no.2
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    • pp.71-74
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    • 1979
  • The growth characteristics of Methylomonas methanolica YUFE 101, isolated from sewage samples, have been studied. conclusions of the study were; (1) Optimum cultivation pH and temperature are 6.3 and 32.5$^{\circ}C$ respectively (2) The specific oxygen uptake rate was 332 $\mu$ι/mg-dry weight/hr. (3) The maximum specific growth rate was 0.19 h $r^{-1}$ and celluar yield was 0.43 g-dry cell/g-methanol in batch culture, (4) The maximum biomass productivity achieved was 0.21 g-dry cel1/ι/hr at a dillution rate of 0.1 h $r^{-1}$ during continuous cultivation. (5) The contents of crude protein and total nucleic acid in the dry cell were 73 % and 12 % respectively.

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Treatment of Aquacultural Recirculating Water by Foam Separation - I. Characteristics of Protein Separation- (포말 분리법을 이용한 양어장 순환수 처리 - I Protein 분리특성 -)

  • SUH Kuen-Hack;LEE Min-Gyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.5
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    • pp.599-606
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    • 1995
  • The feasibility of foam separation to remove protein produced from fish culture water was investigated, By assuming foam separation column as a single well-mixed pool, a simplified model for foam separator conditions was alse studied under the batch operation. The model indicated that the protein removal could be described as a first-order reaction whose rate increases with both superficial air velocity and protein concentration in the bulk solution. from ,the results of an experimental study on the effects of superficial air velocity, the protein concentration, temperature, and pH on protein removal, the superficial air velocity and initial protein concentration in bulk solution were found to be important operational factors. Other experimental results important that foam separator operated under batch conditions would be considered as a completely mixed reactor. The protein removal rate by foam separation process was increased proportionally with the superficial air velocity, and the adsorptive removal rate of protein was found to obey Langmuir adsorption formula. The preposed simplified model was verified with the experimental data obtained from this study. Under the experimental range used, both temperature and pH did not affect the protein removal.

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Characteristics of Batch Cultures and Effects of Various Elicitors on Ginsenoside Production in Suspension Cultures of Panax Ginseng C.A. Meyer (고려인삼세포 현탁배양에서 회분배양 특성 및 Ginsenoside 생산에 대한 다양한 elicitors의 영향)

  • 유병삼;변상요
    • KSBB Journal
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    • v.16 no.6
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    • pp.620-625
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    • 2001
  • This study was examined to investigate the time course behaviors of cell growth and sucrose consumption, and effects of various elicitors on ginsenoside production in batch suspension cultures of Panax ginseng Meyer. Suspended cells reached to the stationary phase at 12 days after innoculation. The maximum cell concentration was 14.7 g-DCW/L at 17 days. The highest cell growth rate was 0.59 g-DCW/L d. The sucrose used as a sole carbon source was hydrolysed to glucose and fructose in 4 days, then quickly utilized until the middle-log phase and consumpted completely at 16 days. Various elicitors were app1ied at 8 days from inoculation which is the middle-log phase. Among the elicitors tested, jasmonic acid was the most efficient to increase the ginseneside production, which was 1.5 times higher than control.

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Optimization of Growth Medium and Fermentation Conditions for the Production of Laccase3 from Cryphonectria parasitica Using Recombinant Saccharomyces cerevisiae

  • Jeong, Yong-Seob;Sob, Kum-Kang;Lee, Ju-Hee;Kim, Jung-Mi;Chun, Gie-Taek;Chun, Jeesun;Kim, Dae-Hyuk
    • Mycobiology
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    • v.47 no.4
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    • pp.512-520
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    • 2019
  • Statistical experimental methods were used to optimize the medium for mass production of a novel laccase3 (Lac3) by recombinant Saccharomyces cerevisiae TYEGLAC3-1. The basic medium was composed of glucose, casamino acids, yeast nitrogen base without amino acids (YNB w/o AA), tryptophan, and adenine. A one-factor-at-a-time approach followed by the fractional factorial design identified galactose, glutamic acid, and ammonium sulfate, as significant carbon, nitrogen, and mineral sources, respectively. The steepest ascent method and response surface methodology (RSM) determined that the optimal medium was (g/L): galactose, 19.16; glutamic acid, 5.0; and YNB w/o AA, 10.46. In this medium, the Lac3 activity (277.04 mU/mL) was 13.5 times higher than that of the basic medium (20.50 mU/mL). The effect of temperature, pH, agitation (rpm), and aeration (vvm) was further examined in a batch fermenter. The best Lac3 activity was 1176.04 mU/mL at 25 ℃, pH 3.5, 100 rpm, and 1 vvm in batch culture.

Genome-wide Analysis and Control of Microbial Hosts for a High-level Production of Therapeutic Proteins

  • Kim, Sung-Geun;Park, Jung-Hwan;Lee, Tae-Hee;Kim, Myung-Dong;Seo, Jin-Ho;Lim, Hyung-Kwon
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2005.06a
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    • pp.230-232
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    • 2005
  • The formation of insoluble aggregation of the recombinant kringle fragment of human apolipoprotein(a), rhLK8, in endoplasmic reticulum was identified as the rate-limiting step in the rhLK8 secretion in Saccharomyces cerevisiae. To analyze the protein secretion pathway, some of yeast genes closely related to protein secretion was rationally selected and their oligomer DNA were arrayed on the chip. The expression profiling of these genes during the induction of rhLK8 in fermentor fed-batch cultures revealed that several foldases including pdi1 gene were up-regulated in the early induction phase, whereas protein transport-related genes were up-regulated in the late induction phase. The coexpression of pdi1 gene increased rhLK8-folding capacity. Hence, the secretion efficiency of rhLK8 in the strain overexpressing pdi1 gene increased by 2-fold comparing in its parental strain. The oligomer DNA chip arrayed with minimum number of the genes selected in this study could be generally applicable to the monitoring system for the heterologous protein secretion and expression in Saccharomyces cerevisiae. With the optimization of fed-batch culture conditions and the alteration of genetic background of host, we obtained extracellular rhLK8 at higher yields than with Pichia pastoris systems, which was a 25-fold increased secretion level of rhLK8 compared to the secretion level at the initiation of this study.

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Optimization for Lacticin SA72 Production by Lactococcus lactis SA72 Isolated from Jeot-gal. (젓갈에서 분리한 Lactococcus lactis SA72에 의한 Lacticin SA72의 생산 최적화)

  • 백현동;구경모;김진곤;이나경
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.46-50
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    • 2003
  • Lactococcus lactis SA72 from Jeot-gal (Korean traditional fermented fish foods) produces lacticin SA72. The influence of several parameters on the fermentative production of lacticin SA72 by Lactococcus lactis SA72 was studied. MRS medium among several media was selected for enhanced bacteriocin production. The mean growth rate and bacteriocin productivity of L. lactis SA72 increased as the initial pH of the media increases. The highest lacticin SA72 activity was detected 3,200 AU/ml at pH 6.0, $32^{\circ}C$, and 1% (inoculum size, v/v) in the jar fermenter. Enhanced production of lacticin SA72 was investigated by a fed-batch cultivation with the intermittent feeding of the concentrated glucose solution. Under the optimized conditions, lacticin SA72 activity finally reached to 6,400 AU/ml.

Enzymatic production of Fructo-oligosaccharides from Sucrose (자당으로부터 프럭토올리고당의 효소적 생산 연구)

  • 신형태;백순용;이수원;서동상;권석태;김종남;임유범;이재흥
    • KSBB Journal
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    • v.17 no.6
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    • pp.555-559
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    • 2002
  • Three different strains of Aureobasidium pullulans were grown in batch cultures to compare their abilities of enzyme production. It was found that specific enzyme activity was the highest with strain ATCC 9348 and the enzyme production was closely coupled to growth. Studies on morphology during the growth of A. pullulans revealed that mycelia cells were dominant at the initial stages of growth. However, yeast-like cells and chlamydospores were dominant in the latter stages of batch culture. The pattern of morphological changes during the growth period was not affected by pH. However, it appears that the ratio of intra- to extracellular enzyme activity tended to increase with fermentation time irrespective of the pH employed, suggesting that the secretion efficiency of intracellular enzyme to broth likely depends on cell morphology Using molasses as a cheap source of sucrose, enzymatic production of fructo-oligosaccharides as a feed additive with A. pullulans cells could be achieved successfully at 55$\^{C}$ and pH 5.5.

Overproduction of Sodium Gluconate Using the Recombinant Aspergillus niger (재조합 Aspergillus niger에 의한 글루콘산나트륨의 산업적 생산)

  • 이선희;이현철;김대혁;양문식;정봉우
    • KSBB Journal
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    • v.13 no.2
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    • pp.214-219
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    • 1998
  • Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400 $\mu$m ml$^-1$ of hyg.B while the minimal media inhibited mycelial growth completely at 200 $\mu$m ml$^-1$ of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.

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Optimization of Medium and Fermentation Conditions for Mass Production of Bacillus licheniformis SCD121067 by Statistical Experimental Design (Bacillus licheniformis SCD121067 균체 생산성 증가를 위한 통계적 생산배지 및 발효조건 최적화)

  • Jeong, Yoo-Min;Lee, Ju-Hee;Chung, Hea-Jong;Chun, Gie-Taek;Yun, Soon-Il;Jeong, Yong-Seob
    • KSBB Journal
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    • v.25 no.6
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    • pp.539-546
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    • 2010
  • In this work, mass production of Bacillus licheniformis SCD121067 through medium optimization by statistical experimental method was studied. First, galactose, yeast extract and potassium phosphate dibasic were selected as carbon, nitrogen and phosphate sources for mass production of B. licheniformis SCD121067 by using one factor at a time method. Second, according to the result of Plackett-Burman experimental design, key factors was yeast extract and $K_2HPO$. Finally, the response surface methodology was performed to obtain the optimum concentrations of two selected variables. The optimized medium composition consisted of 20 g/L galactose, 36 g/L yeast extract, 0.41 g/L $K_2HPO4$, 0.25 g/L $Na_2CO_3$, 0.4g/L $MgSO_4$ and 0.01g/L $CaCl_2$. Dry cell weight (15.4 g/L) by optimum production medium were increased 10 times, as compared to that determined with basic production medium (1.5 g/L). Fermentation conditions were examined for the mass production of B. licheniformis. The effect of temperature, agitation speed, pH and aeration rate on the mass production of B. licheniformis were also studied in a batch fermenter which was carried out in a 2.5 L bioreactor with a working volume of 1.5 L containing optimized production medium. As a result, dry cell weight of batch culture was 30.7 g/L at $42^{\circ}C$, 300 rpm, pH 8.0 and 2 vvm.