• KSBB Journal
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• v.10 no.4
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• pp.374-385
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• 1995

Nutritional requirements and cultural conditions for the production of extracellular gelatin-degrading proteolytic enzyme by Bacillus subtilis B0021 were investigated. Optimum carbon source for proteolytic enzyme production was salicin, but it was substituted by glucose for economical reason. The fermentation medium giving a maximum proteolytic enzyme activity was consisted of 1.5%(w/v) glucose, 2.5%(w/v) yeast extract, and 0.001%(w/v) manganese sulfate and 0.002%(w/v) ferrous sulfate. Proteolytic enzyme activity of B. subtilis B0021 was completely inhibited by 0.5%(w/v) tannic acid. Initial pH was optimal at 7.0 and the enzyme activity in the flask culture usually reached a maximal level after 36 hours of fermentation at $30^{\circ}C$. In the $5\ell$ fermentor fermentation at $30^{\circ}C$, enzyme activity was maximum at 36 hour of cultivation but after this enzyme activity was decreased rapidly. Initial viscosity of 45%(w/v) gelatin(2,800mPas) was decreased rapidly to 96%(mPas) after hydrolysis for 4hr at $40^{\circ}C$ by crude enzyme of B. subtilis B0021.

• KSBB Journal
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• v.25 no.6
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• pp.520-526
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• 2010

In this work we have investigated the production of catalase from Bacillus sp. strains, which were screened and identified from soil. These strains were cultivated in shaking flasks with tryptic soy broth (TSB) at $30^{\circ}C$ and 200 rpm. Effects of the temperature and pH on the stability of the native catalase and whole cell viability were studied in the temperature range of $25-60^{\circ}C$ and the pH range of 7-13. Korean natural zeolite was added to culture medium and mixed with microorganisms for 24 hours. The native catalase maintained its activity over $50^{\circ}C$. The enzyme acitiviy of the catalase from Bacillus flexus BKBChE-3 was highest among the Bacillus sp. strains studied. Bacillus flexus BKBChE-3 and immobilized Bacillus cells have survived under extreme conditions of over $50^{\circ}C$ and pH 12. 60 mL of 10.5 mM $H_2O_2$ solution were entirely removed within 1 hour with catalase produced from Bacillus sp. on the flask. When Bacillus cells were immobilized on Korean natural zeolite, colony forming unit of Bacillus flexus BKBChE-3 was increased and high efficiency of hydrogen peroxide removal was observed.

• Journal of Korean Society of Forest Science
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• v.41 no.1
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• pp.1-6
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• 1979

1. The study was conducted on the optimum condition of the treated substrate with dilute sulphuric acid solution and cellulase for saccharification. The wood (saw dust) of Alnus hirsuta Rupr. (10~15 years) was treated with 0.3%, 0.6%, 0.9%, 1.2%, 1.5%, $H_2SO_4$ solution at $1.5kg/cm^2$ for 15min., 30min., 45min., and 60min., followed by thermal treatment at $190^{\circ}C$ for 30min., and screening with 60 mesh sieve, after which to 0.5 grams of each sample was added 0.5ml cellulase solution, and 50ml 0.1M acetic acid buffer solution (pH 5.0), after incubating at $40^{\circ}C$ for 96hr. 2. The crude cellulase of Trichoderma viride Perx. ex. Fr. SANK 16374 was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfite. 3. Reducing sugar was determined by the 3.5-dinitrosalicylic acid (DNS) method. 4. The reducing sugar was increased with increase of the sulphuric acid concentration and saw dust was treated with 1.5% $H_2SO_4$ solution at $1.5kg/cm^2$ for 45min. showed the best saccharification (16.0%). The reducing sugar formation did not show statistically significant in 5% levels by thermal treatment time 45min. and 60min. 5. The substrate for cellulase which was treated with 0.9% $H_2SO_4$ solution at $1.5kg/cm^2$ for 60min. showed the best reducing sugar formation (23.6%). And did not show significant difference in 5% levels at 0.9%, 1.2%, and 1.5% $H_2SO_4$ solution.

• Journal of Life Science
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• v.31 no.1
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• pp.90-99
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• 2021

In this study, to reduce the production cost of poly(3-hydroxyalkanoates) (PHA), optimal cell growth and PHA biosynthesis conditions of the isolated strain Pseudomonas sp. EML8 were established using waste frying oil (WFO) as the cheap carbon source. Gas chromatography (GC) and GC mass spectrometry analysis of the medium-chain-length PHA (mcl-PHAWFO) obtained by Pseudomonas sp. EML8 of WFO indicated that it was composed of 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate, and 16.58 mol% 3-hydroxvdodecanoate monomers. When Pseudomonas sp. EML8 were culture in flask, the maximum dry cell weight (DCW) and the mcl-PHAWFO yield (g/l) were showed under WFO (20 g/l), (NH4)2SO4 (0.5 g/l), pH 7, and 25℃ culture conditions. Based on this, the highest DCW, mcl-PHAWFO content, and mcl-PHAWFO yield from 3-l-jar fermentation was obtained after 48 hr. Similar results were obtained using 20 g/l of fresh frying oil (FFO) as a control carbon source. In this case, the DCW, the mcl-PHAFFO content, and the mcl-PHAFFO yields were 2.7 g/l, 62 wt%, and 1.6 g/l, respectively. Gel permeation chromatography analysis confirmed the average molecular weight of the mcl-PHAWFO and mcl-PHAFFO to be between 165-175 kDa. Thermogravimetric analysis showed decomposition temperature values of 260℃ and 274.7℃ for mcl-PHAWFO and mcl-PHAFFO, respectively. In conclusion, Pseudomonas sp. EML8 and WFO could be suggested as a new candidate and substrate for the industrial production of PHA.

• KSBB Journal
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• v.23 no.3
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• pp.205-212
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• 2008

The efficient soluble expression of human epidermal growth factor (hEGF) was achieved by using functional fusion partners in cytoplasm and periplasm of Escherichia coli (E. coli). hEGF was over-expressed in inactive inclusion body form in cytoplasm of E. coli due to improper disulfide bond formation and hydrophobic interaction, yielding about 5.9 mg/L in flask culture. Six functional fusion partners were introduced by linking to N-terminal part of hEGF gene for the high-level expression of soluble and active hEGF in cytoplasm and peri plasm region. Three fusion partners for cytoplasmic expression such as acidic tail of synuclein (ATS), thioredoxin (Trx) and lipase, and three fusion partners for periplasmic expression such as periplasmic cystein oxidoreductases (DsbA and DsbC) and maltose binding protein (MBP) were investigated. hEGF fused with ATS and DsbA showed over 90% of solubility in cytoplasm and periplasm, respectively. Especially DsbA was found to be an efficient fusion partner for soluble and high-level expression of hEGF, yielding about 18.1 mg/L and three-fold higher level compared to that of insoluble non-fusion hEGF in cytoplasm. Thus, heterologous proteins containing complex disulfide bond and many hydrophobic amino acids can effectively be produced as an active form in E. coli by introducing a suitable peptide or protein.

• Journal of Life Science
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• v.25 no.9
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• pp.1027-1035
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• 2015

A strain GP32 which produces a highly viscous extracellular polysaccharide was conducted with soil samples and identified as Pseudomonas species. The culture flask conditions for the production of extracellular polysaccharide by Pseudomonas sp. GP32 were investigated. The most suitable carbon and nitrogen source for extracellular polysaccharide production were galactose and (NH₄)₂SO₄. The optimum carbon/nitrogen ratio for the production of extracellular polysaccharide was around 50. The optimum pH and temperature for extracellular polysaccharide production was 7.5 and 32℃, respectively. In batch fermentation using a jar fermentor, the highest extracellular polysaccharide content (15.7 g/l) was obtained after 70 hr of cultivation. The extracellular polysaccharide produced by Pseudomonas sp. GP32 (designated Biopol32) was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation, and gel permeation chromatography. Biopol32, which has an estimated molecular weight of over 3×10⁷ datons, is a novel polysaccharide derived from sugar components consisting of galactose, glucose, gulcouronic acid and galactouronic acid in an approximate molar ratio of 1.85 : 3.24 : 1.00 : 1.42. The solution of Biopol32 showed non-Newtonian characteristics. The viscosity of Biopol32 exhibited appeared to be higher at all concentration compared to that of zooglan from Zoogloea ramigera. An analysis of the flocculating efficiency of Biopol32 in industry wastewater (food, textile, and paper wastewater) revealed chemical oxygen demand (COD) reduction rates 58.4-67.3% and suspended solid (SS) removal rates 82.6-91.3%. Based on these results, Biopol32 is a possible candidate for industrial applications such as wastewater treatment.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.317-323
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• 2014

Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

• Journal of Life Science
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• v.29 no.8
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• pp.861-870
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• 2019

This study was undertaken to establish optimum medium compositions for cost-effective mannitol production by Leuconostoc mesenteroides SRCM201425 isolated from kimchi. L. mesenteroides SRCM21425 from kimchi was selected for efficient mannitol production based on fructose analysis and identified by its 16S rRNA gene sequence, as well as by carbohydrate fermentation pattern analysis. To enhance mannitol production by L. mesenteroides SRCM201425, the effects of carbon, nitrogen, and mineral sources on mannitol production were first determined using Plackett-Burman design (PBD). The effects of 11 variables on mannitol production were investigated of which three variables, fructose, sucrose, and peptone, were selected. In the second step, each concentration of fructose, sucrose, and peptone was optimized using a central composite design (CCD) and response surface analysis. The predicted concentrations of fructose, sucrose, and peptone were 38.68 g/l, 30 g/l, and 39.67 g/l, respectively. The mathematical response model was reliable, with a coefficient of determination of $R^2=0.9185$. Mannitol production increased 20-fold as compared with the MRS medium, corresponding to a mannitol yield 97.46% when compared to MRS supplemented with 100 g/l of fructose in flask system. Furthermore, the production in the optimized medium was cost-effective. The findings of this study can be expected to be useful in biological production for catalytic hydrogenation causing byproduct and additional production costs.