• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.371-374
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• 2002

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)［P(3HB/V)］, by fed-batch culture of recombinant Escherichia coli harboring a plasmid containing the Alcaligenes latus polyhy-droxyalkanoate (PHA) biosynthesis genes, was examined in two pilot-scale fermentors with air supply only, In a 30 L fermentor having a XLa value of 0.11 S­$^1$, the final P(3HB/V) concentration and the P(3HB/V) content obtained were 29.6 g/L and 70.1 wt%, respectively giving a productivity of 1.37 g P(3HB/V)/L-h. In a 300 L fermentor having a XLa of 0.03 S­$^1$, the P(3HB/V) concentration and the P(3HB/V) content were 20.4 g/L and 69 wt%, respectively giving a productivity of 1.06g P(3HB/V)/L-h. These results suggest that economical production of P(3HB/V) is possible by fed-batch culture of recombinant E. coli in a large-scale fermentor having low KLa value.

• Journal of Life Science
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• v.23 no.7
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• pp.886-892
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• 2013

We investigated ethanol production from glycerol after screening of the yeast Pachysolen tannophilus ATCC 32691. For yeast to produce ethanol form glycerol, it is important that aeration is finely controlled. Therefore, we attempted to produce ethanol using a surface-aerated fermentor. When 880 ml of YPG medium (1% yeast extract, 2% peptone, 2% glycerol) was used to produce ethanol, the optimal aeration conditions for ethanol production were a surface aeration rate and agitation speed of 500 ml/min and 300 rpm, respectively. In a fed-batch culture, the maximum ethanol production and the maximum ethanol yield from glycerol (Ye/g) was 5.74 g/l and 0.166, respectively, after 90 hr using the surface-aerated fermentor.

• KSBB Journal
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• v.13 no.2
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• pp.187-194
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• 1998

For mass production of polyhydroxyalkanoates (PHA), high cell density cultures of Alcaligenes eutrophus by fed-batch culture under phosphate-limitation condition has been investigated. PHA accumulation by the regulation by the regulation of initial phosphate concentration could be automatically induced, and high density cell culture above 200 g/L also could be successfully produced. The production of Poly-$\beta$-hydroxybutyrate (PHB) and dry cell weight increased with increasing the initial phosphate concentration. When the initial concentrations of phosphate were in the ranges of 1.5~4.5 g-PO$_4$/L, PHB and dry cell weight obtained were 83~266 g/L and 61~216 g/L, respectively, and PHB productivity was in the ranges of 1.35~3.10 g/L.h. When a mixture of glucose and propionic acid is used as carbon sources, poly(3-hydroxybutyrate-co-poly-3-hydroxyvalerate), P(3HB-co-3HV), could be also successfully produced under phosphate limitation condition. When the mole ratio of propionic acid to glucose in the feeding solution is 0.22, a final dry cell weight of 150 g/L and a P(3-HB-co-3HV) of 90 g/L were produced. Morphological changes and size distribution of PHB granules synthesized in A. eutrophus under phosphate-limitation condition are determined by TEM during the course of fed-batch. Mean granule diameters of PHB produced are in the range of 0.36~0.39 $\mu$m, and mean cell size was elongated from 0.54~0.59 $\mu$m$\times$ 1.3~1.5 $\mu$m to 0.83~0.89 $\mu$m $\times$2.0~2.3 $\mu$m. Phosphate concentration in media did not affect size distribution of PHB granule and cell.

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.343-347
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• 1997

Effect of culture conditions on the fermentation of wheat flour solution by mixed lactic acid bacteria of Lactobacillus brevis, L. fermentum and L. plantarum was investigated. The optimum temperature for the fermentation of wheat flour solution was $35^{\circ}C$ because pH decreased the lowest value and TTA (total titrable acidity) increased the highest value at this temperature. In aerobic condition, fermentor was purged with air at 1.0 vvm and was purged with nitrogen gas at 1.0 vvm in anaerobic condition. The decrease of pH and the increase of TTA in aerobic condition were higher than those in anaerobic condition. In aerobic condition, the optimum condition of oxygen supply was found to be oxygen transfer rate coefficient of $60\;hr^{-1}$ which corresponded to agitation speed of 250 rpm in a 5 L fermentor. Repeated fed-batch cultures were performed using pH-stat in order to increase the productivity of fermented wheat flour. With increasing the repeated fraction of culture volume, mean cycle time increased but maximum operation time decreased. However, the volume of produced broth per culture volume per time and total volume of produced broth per culture volume were maximum at the repeated fraction of culture volume of 20%. In a repeated fed-batch fermentation of wheat flour solution using mixed lactic acid bacteria, the culture condition was optimum at temerature of $35^{\circ}C$, aeration rate of 1.0 vvm, oxygen transfer rate coefficient of $60\;hr^{-1}$, and repeated fraction of culture volume of 20%.

• Microbiology and Biotechnology Letters
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• v.29 no.4
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• pp.206-211
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• 2001

In order to overexpress constitutively the Kluyveromyces marxianus exoinulinase gene (INUI) in Saccharomyces cerevisiae, four episomal expression systems employing GAPDH, ADHI, PGK and ENOI promoters were constructed as p YIGP aADHI -INU, pPGK-INU, and pENOI- INU plasmids respectively, When S cereviais transformants harboring each plasmid were batchwisely cultivated in the fermentor containing 5% glucose medium no significant differences in the cell growth are observed How- ever the experession level of exoinulinase and plasmid stability showed a strong dependency on the promoter employed. The expression levels of exoinulinase were about 1.70 unit/ml for GAPDH promoter 1.67 unit/ml for PGK promoter, 1.29 unit /ml for ADH1 promoter, and 0.80 unit/ml for ENOl promoter. The plasmid stabilites were maintaines above 80% in all experession systems. except the GAPDH promoter system of 55%, Based on the plas- mid stability and expression level of exoinulinase the ADHl and PGK promoter system were selected for the fed - batch culture to overproduce exoinulinase By the intermittent feeding of yeast extract and glucose, both promoter systems gave the cell concentration of about 30 g-dry cell weight/1 byt the maximal exoinulinase activity of 3.70 unit/ml and plasmid stability of 96% in the ADH1 promoter were higher than those (2.70 unit/ml, 80%) of PGK sys- tem Taking into account the plasmid stability and extended culture time the ADH1 promoter systems would be the most feasible expression systems for the constitutive overproduction of exoinulinase through high cell-density fed- batch cultures using non-selective rich medium.

• Microbiology and Biotechnology Letters
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• v.35 no.3
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• pp.226-230
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• 2007

In order to achieve high-level expression of interferon-${\alpha}1$ (IFN-${\alpha}1$) during fed-batch fermentation of recombinant E. coli, effects of oxygen supply and induction temperature on the expression of recombinant proteins were evaluated. Supplementation of oxygen and its transfer into cells is one of the most important parameters involved in the design and operation of mixing-sparging equipment for bioreactors. Generally, higher oxygen supply stimulates cell growth of aerobic microorganism and consequently the amount of products is increased. In this study, the optimum aeration strategy for the higher production of IFN-${\alpha}1$ during fed-batch fermentation of recombinant E. coli was surveyed. The growth of the cells was also monitored with four different concentrations of dissolved oxygen (DO; limiting, 20%, 35%, 50%) conditions. The DO was controlled by varying aeration rates of air and pure oxygen. Oxygen uptake rate (OUR) and specific oxygen uptake rate (SOUR) were evaluated and compared for the enhanced growth and induction of the cells and IFN-${\alpha}1$, respectively. We confirmed that increased DO by additional oxygen supply, up to 35%, can improve the production of IFN-${\alpha}1$ during the fermentation.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.263-268
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• 1989

To maximize the performance of recombinant cell fermentation process through optimizing environmental conditions, the production of invertase from recombinant Saccharomyces cerebisiae Containing SUC2 gene was studied as a model. The recombinant cells showed biphasic growth on glucose. Since the promoter of the SUC2 is regulated by the concentration of glucose in the medium, expression of invertase by recombinant yeast began when the glucose concentration decreased in a range of 0.25-0.4 g/L during the batch culture. Plasmid segregation occured frequently during glucose fermentation, and infrequently during ethanol oxidation. A rapid appearance of invertase activity with glucose was observed under nonaerated condition, and the maximum specific invertase activity was about 1.5 times as high as under aerobic condition, In fed batch culture, when n low level of glucose was continuously supplied to the tormentor after the time of glucose depletion during growth phase, specific and total invertase activity increased about 1.7 and 2.9 fold, respectively, in a batch culture.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.399-405
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• 1986

Corn starch was saccharified without cooking in an agitated bead reaction system. Uncooked corn starch was effectively hydrolyzed even at the concentration as high as 39%(w/v). After 24 hours. the extent of saccharification reached at 92%, which corresponds glucose concentration of 425g/L. Fed-batch feeding of starch was more effective than batch feeding for saccharification of uncooked corn starch. The composition of hydrolysated of uncooked starch was analyzed. which was composed of 95% glucose, 0.7% of maltose, and 4.5% of high saccharide, similar with that of cooked starch. The hydrolysate can be successfully utilized for HFCS manufacture. The starch liquefying and saccharifying enzyme was relatively stable even be the physical impact of the attrition-milling media. The enzyme stabilizer, $Ca^{++}$, played an essential role in preventing the enzyme deactivation caused by the physical impact.

• KSBB Journal
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• v.18 no.2
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• pp.155-160
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• 2003

The effect of dissolved oxygen(DO) on microbial transglutaminase(mTG) production by Streptoverticillium morbaraense was studied in on-line computer controlled fermentation system. In order to control dissolved oxygen during fermentation, the agitation speed and aeration rate of 2.5 L fermenter ranged from 260 to 360 rpm and 0.3 to 3.9 L/min, respectively. The maximum microbial transglutaminase production was obtained at controlled 20% of dissolved oxygen among the various dissolved oxygen controlled batch cultures tested. The production of microbial transglutaminase at controlled 20% of dissolved oxygen was about 2.12 U/mL which was 1.1 times higher than that obtained in batch culture without control of dissolved oxygen. Also, the highest microbial transglutaminase production was obtained in fed-batch cultures in which dissolved oxygen was controlled at 20%, and it was improved almost 1.3 times in comparison with that without control of dissolved oxygen. Maximal dry cell weight and microbial transglutaminase production were 13.2 g/L and 2.6 U/mL, respectively. Finally, it was also found that fed-batch fermentation at controlled 20% of dissolved oxygen showed a good performance for the microbial transglutaminase production by on-line computer controlled fermentation system which may be generally applicable to other microbial cultures.

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1938-1944
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• 2008

The procpb gene encoding human procarboxypeptidase B (proCPB, GeneBank access code AJ224866) was cloned and its Pichia expression plasmid, $pPIC9{\alpha}$/hproCPB (9.2 kb), was constructed, in which procpb was under the control of the AOXl promoter and connected to the downstream of the mating factor ${\alpha}$-1 ($MF{\alpha}1$) signal sequence. The plasmid was linearized by digestion with Sacl, and integrated into the genome of P. pastoris strain GS115. By culturing of Pichia transformant on methanol medium, the human proCPB was successfully expressed and secreted into the culture supernatant. Moreover, Western blot analysis of the extracellular proteins showed proCPB bands clearly at a molecular mass of 45 kDa, confirming the expression of proCPB with its right size. The CPB activity reached about 3.5 U/ml and 12.7 U/ml in the flask and fermentor batch cultures of Pichia transformant, respectively. No CPB enzyme activity was found in the intracellular fraction. When the fed-batch cultivation was performed with methanol and glycerol mixture as a feeding medium, the extracellular CPB activity was increased to 42.0 U/ml, which corresponds to a 3.3-fold higher level of CPB activity than that of batch culture. The $K_m$ and $k_{cat}$ values of recombinant human CPB enzyme for hippuryl-$_L$-Arg as a substrate were estimated to be 0.16 mM and $11.93\;sec^{-1}$, respectively.