• Journal of Life Science
• /
• v.12 no.3
• /
• pp.242-249
• /
• 2002

The optimal culture conditions in 500$m\ell$ flask suture, 5$\ell$ jar fermenter and 2,000 $\ell$ large stale culture were investigated to maximize the production of antibiotic on Rhizoctonia solani AC4, the causal agent of vegetables damping-off, by the strain Bacillus ehimensis YJ-37. Starch (1.5%) as a carbon source, peptone (0.6%) as a nitrogen source and MgC1$_2$(0.15%) as a metal ion in the medium containing N $a_2$HP $O_4$(0.3%) showed the highest production of the antibiotic(s) in a rotary shake (200 rpm). Optimal initial pH of the culture medium, culture temperature and culture time for the antibiotic(s) production were pH 8.0, 32$^{\circ}C$ and 54hrs, respertively. Under the optimal renditions in flask culture, cell growth and antifungal activity (clear zone size) were 1.42 $\times$ 10$^{8}$ cfu/$m\ell$ (21g-DCW/ $\ell$) and 13.9 mm, respectively. In 5 $\ell$ jar fermenter (medium 3 $\ell$), optimal air flow, agitation speed and culture time for the antibiotic(s) production were 2 vvm, 200 rpm and 48hrs, respectively. Under the optimal conditions in 5 $\ell$ jar fermenter, tell growth and antifungal activity were 2.06 $\times$ 10$^{8}$ cfu/$m\ell$ (30g-DCW/ $\ell$) and 13.4 mm, respectively. Under the culture conditions of air flow (30 vvm) and agitation speed (200 rpm) at 32$^{\circ}C$ for 10 days in 2,000 $\ell$ large scale culture (medium 1,800 $\ell$, pH 8.0), cell growth and antifungal activity were 0.81$\times$10$^{8}$ cfu/$m\ell$ (12g-DCW/ $\ell$) and 8.6 mm, respectively.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.378-381
• /
• 2001

In the batch culture experiments, the addition of $MnSO_4$, was examined in flask culture, and then the optimal amounts of $MnSO_4$ was investigated in 2.5- L jar-fermenter. As a results, 0.005% $MnSO_4$ was found to enhance the ARA yield of 1.14- fold. Also the addition of $KH_2PO_4_4$ was investigated in 2.5-L jar-fermenter and the ARA yield was enhanced 1.20-fold. Fed batch culture study shown a relatively high productivity of cell mass (62.1 g/L) and ARA content (12.0 g/L) when 14% ammonia solution were alternatively used to control the pH and nitrogen source in the cultivation period.

• KSBB Journal
• /
• v.14 no.2
• /
• pp.241-247
• /
• 1999

Experimental studies were carried out to optimize the culture conditions of Corynebacterium diphtheriae for the production of diphtheria toxin. A new media which does not contain any meat digest products was selected. The main ingredient of new medium was enzymatic digests of casein known as NZ-Case. In fermenter experiments, the toxin production was increased with the increase of cell growth. The optimum initial pH of media, air flow rate and agitation speed were 7.0, 0.22, vvm and 400 rpm, respectively. The contents of iron and calcium-phosphate precipitate were important for maximal cell growth and toxin production. The optimum concentration of iron was 0.3 mg/L and calcium-phosphate precipitate could serve in gradual supply of iron to maintain the optimal culture condition which is required for enhanced yield of toxin production. In potency test, the potency of toxoid from fermentor culture was higher than that from static culture. When diphtheria toxin is produced by fermentor culture, it is possible to produce higher levels of toxin and better toxoid quality in terms of safety, yield, productivity and immunity.

• Asian-Australasian Journal of Animal Sciences
• /
• v.13 no.10
• /
• pp.1399-1406
• /
• 2000

A continuous culture study was conducted to determine the impact of ruminal degradable soy protein (S-RDP) level and dilution rate (D) on growth of ruminal non-structural carbohydrate-fermenting microbes. Corn starch, urea and isolated soy protein (ISP) were used to formulate three diets with S-RDP levels of 0, 35 and 70% of total dietary CP. Two Ds were 0.03 and $0.06h^{-1}$ of the fermenter volume in a single-effluent continuous culture system. As S-RDP levels increased, digestibilities of dietary dry matter (DM), organic matter (OM) and crude protein (CP) linearly (p=0.001) decreased, whereas digestion of dietary starch linearly (p=0.001) increased. Increasing D from 0.03 to $0.06h^{-1}$ resulted in decreased digestibilities of dietary DM and OM, but had no effect on digestibilities of dietary starch (p=0.77) and CP (p=0.103). Fermenter pH, the concentration of volatile fatty acids (VFA) and daily VFA production were unaffected (p=0.159-0.517) by S-RDP levels. Molar percentages of acetate, propionate and butyrate were greatly affected by S-RDP levels (p=0.016-0.091), but unaffected by D (p=0.331-0.442). With increasing S-RDP levels and D, daily bacterial counts, daily microbial N production (DMNP) and microbial efficiency (MOEFF; grams of microbial N produced per kilogram of OM truly digested) were enhanced (p=0.001). The increased microbial efficiency with increasing S-RDP levels is probably the result of peptides or amino acids that served as a stimulus for optimal protein synthesis. The quantity of ruminal degradable protein from soy proteins required for optimum protein synthesis of non-structural carbohydrate-fermenting microbes appears to be equivalent to 9.5% of dietary fermented OM.

• KSBB Journal
• /
• v.19 no.1
• /
• pp.78-82
• /
• 2004

Experiments were carried out to select an organic nitrogen source and optimize the culture conditions for the production of arachidonic acid by Mortierella alpina DSA-12. Corn steep powder(CSP) was selected as an organic nitrogen source based on arachidonic acid production and raw material price. The optimum C/N ratio was in the range of 15 to 17 with the medium containing glucose as carbon source and CSP as nitrogen source. The optimum culture conditions for arachidonic acid production showed 500 rpm agitation and 25$^{\circ}C$ culture temperature at 0.5 vvm aeration. Under the optimum conditions, the concentration of cell, total lipid and arachidonic acid were 21.8 g/L, 10.2 g/L and 3.70 g/L, respectively, from 50 g/L glucose and 18 g/L CSP. In the 500 L fermenter with 0.5 vvm aeration and 200 rpm agitation, the concentration of cell, total lipid and arachidonic acid were 19.8 g/L, 9.1 g/L and 3.67 g/L, respectively, from 50 g/L glucose and 18 g/L CSP. This result showed that an arachidonic acid production could be possible with a bench-scale fermenter using corn steep powder as a nitrogen source.

• KSBB Journal
• /
• v.31 no.2
• /
• pp.105-112
• /
• 2016

Ferritin is known to regulate iron metabolism and maintain iron in a variety of the eukaryotic organisms. The region encoding the mature ferritin (0.47 kb, H-type) of Periserrula leucophryna was amplified using the designed primers including restriction enzyme site and termination codon and subcloned in frame to the pRBAS secretion vector containing the signal sequence, RBS, and promoter of amylase gene (E. coli-Bacillus shuttle vector), resulting in recombinant pRBAS-PLF vector. Recombinant ferritin (18 kDa) was correctly processed and secreted from Bacillus subtilis LKS strain harboring the pRBAS-PLF vector and quantitatively analyzed by SDS-PAGE and western blot, respectively. Secretion of the ferritin was optimized by culture conditions (host, medium, temperature, nitrogen source) in 3 L batch culture and 5 L jar fermenter. Finally. the ferritin was largely produced using 50 L fermenter as the following conditions; at $30^{\circ}C$, 150 rpm, 1 vvm in Bacillus subtilis LKS using PY medium. The secreted ferritin was maximally measured (approximately 177.6 ug/ml) when the cell density reached to 14.4 at $OD_{600}$ (20 h incubation). The iron binding activity was confirmed by Perls' staining in 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in the culture broth after secretion. Biologically, the culture broth and powder type containing ferritin were tested for possibility as feed additive in chicken broiler. As a result, the ferritin stimulated the growth of chick broil and improved feed efficiency and production index.

• Applied Biological Chemistry
• /
• v.48 no.3
• /
• pp.207-211
• /
• 2005

The metabolites released from cultures of rhizosphere bacteria can inhibit plant growth. Bacillus subtilis IJ-31 inhibited plant growth by the production of hydrocinnamic acid (HCA). The production of HCA by plant-growth inhibiting rhizobacterium B. subtilis IJ-31 was optimized. $90.5\;{\mu}g/ml$ of HCA was obtained under the condition of 1% rice bran as carbon source, 0.5% tryptone as nitrogen source, 0.1% $ZnCl_2$ as metal source at $37^{\circ}C$ for 60 h (pH 7.0). The optimal condition for the HCA production by B. subtilis IJ-31 in the jar fermenter was established using response surface methodology (RSM) of statistical analysis system(SAS) program. The production of HCA by B. subtilis IJ-31 in the jar fermenter culture reached $102.99\;{\mu}g/ml$ when 2.24% soil extracts was added and agitation speed was 290 rpm under the same condition. And the experimental value of HCA production is $102.5\;{\mu}g/ml$ in the same culture condition. The production of HCA by B. subtilis IJ-31 is higher as 12% than that from the flask culture.

• KSBB Journal
• /
• v.24 no.3
• /
• pp.303-311
• /
• 2009

A scaling up study for the exopolysaccharide (EPS) production by submerged culture of Ganoderma lucidum was carried out in jar fermenter systems (2.6, 20 and 75 L) under bi-staged pH process. Profiles of dissolved oxygen (DO) and volumetric coefficient of oxygen transfer ($k_La$) as a function of operating variables (agitation speed and aeration rate) was investigated, and a correlation between $k_La$ and operating variables was analysed statistically. Under bi-staged pH process, no limitation of DO was observed at agitation speeds tested in the range of 200 and 600 rpm, and the highest EPS production was obtained at the level of DO of $40{\sim}80%$. From the regression analysis, the relation between $k_La$, gas velocity (Vs), stirrer speed (N) and impeller diameter (Di) could be expressed as : $$k_La=0.555{\times}Vs^{0.42}{\times}(N^3{\times}Di^2)^{0.33}\;(R^2=0.925,\;p<0.05)$$ It was found that under 2.6 L jar fermenter, the optimum agitation speed and aeration rate was 400 rpm and 1 vvm, respectively, obtaining the EPS production of 15.43 g/L. Under the submerged cultivation of G. lucidum in jar fermenters of $2.6{\sim}75\;L$, the similar EPS yields at each fermenter were achieved during scaling up based on $k_La$, and $k_La$ value for maximum EPS production was $85.4{\pm}26.70\;h^{-1}$.

• Journal of Korea Technical Association of The Pulp and Paper Industry
• /
• v.29 no.1
• /
• pp.43-51
• /
• 1997

The various culture conditions of Trichoderma viride(ATCC 3454) and Phanerochaete chrysosporium(ATCC 26921) with glucose-pepton medium, Mandels medium, YMG medium for wood degradable enzyme were examined. Mycellium of the two species grew profusely on glucose-pepton medium. Maximum fungal growth was observed about 10days. But CMCase, Fpase, laccase activity in the culture medium with glucose-pepton was not detected. When grown in fermenter culture using Mandels medium, Trichoderma viride produced CMCase and Fpase. Its CMCase activity was 0.15 lU/ml and Fpase activity was 0.3 IU/ml within about 4-6days. Phanerochaete chrysosporium grown in a YMG medium gave the best enzyme activity when they were grown under stationary culture with an atmosphere of 100% oxygen. Levels of laccase activity of 3.0 mull were achieved in stationary culture under 100% oxygen. The enzyme condensation by ultrafiltration method caused a 2-fold(cellulase) and 6-fold(laccase) as compared to control activity.

• 한국생물공학회:학술대회논문집
• /
• 2005.10a
• /
• pp.312-315
• /
• 2005

A process for efficient recycle fed-batch culture was carried out to increase cell mass and spore production by Lactic acid bacteria isolated from Kimchi. A large quantity of cell mass obtained by feeding concentration of sugar in recycle fed-batch culture. When the high density of salt was created that the cell mass was come-down. In this study, cultured in different feeding concentration of sugar conditions. Lactic acid bacteria by recycle fed-batch culture was investigated in 2L working volume of fermenter, obtained the maximum cell mass was 15.17g/L.