• Journal of Microbiology and Biotechnology
• /
• v.27 no.5
• /
• pp.947-955
• /
• 2017

Herbicidin A is a potent herbicide against dicotyledonous plants as well as an antibiotic against phytopathogens. In this study, fermentation parameters for herbicidin A production in submerged culture of Streptomyces scopuliridis M40 were investigated. The herbicidin A concentration varied with the C/N ratio. High C/N ratios (>4) resulted in a herbicidin A production of more than 900 mg/l, whereas maximally 600 mg/l was obtained at ratios between 1 and 3.5. In 5-L batch fermentation, there was a positive correlation between the oxygen uptake rate (OUR) and herbicidin A production. Once the OUR increased, the substrate consumption rate increased, leading to an increase in volumetric productivity. Mechanical shear force affected the hyphal morphology and OUR. When the medium value of hyphal size ranged from 150 to $180{\mu}m$, high volumetric production of herbicidin A was obtained with OUR values >137mg $O_2/l{\cdot}h$. The highest herbicidin A concentration of 956.6 mg/l was obtained at 500 rpm, and coincided with the highest relative abundance of hyphae of $100-200{\mu}m$ length and the highest OUR during cultivation. Based on a constant impeller tip speed, which affects hyphal morphology, herbicidin A production was successfully scaled up from a 5-L jar to a 500-L pilot vessel.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.7
• /
• pp.939-946
• /
• 2012

Antibiotic production with Streptomyces sindenensis MTCC 8122 was optimized under submerged fermentation conditions by artificial neural network (ANN) coupled with genetic algorithm (GA) and Nelder-Mead downhill simplex (NMDS). Feed forward back-propagation ANN was trained to establish the mathematical relationship among the medium components and length of incubation period for achieving maximum antibiotic yield. The optimization strategy involved growing the culture with varying concentrations of various medium components for different incubation periods. Under non-optimized condition, antibiotic production was found to be $95{\mu}g/ml$, which nearly doubled ($176{\mu}g/ml$) with the ANN-GA optimization. ANN-NMDS optimization was found to be more efficacious, and maximum antibiotic production ($197{\mu}g/ml$) was obtained by cultivating the cells with (g/l) fructose 2.7602, $MgSO_4$ 1.2369, $(NH_4)_2PO_4$ 0.2742, DL-threonine 3.069%, and soyabean meal 1.952%, for 9.8531 days of incubation, which was roughly 12% higher than the yield obtained by ANN coupled with GA under the same conditions.

• Microbiology and Biotechnology Letters
• /
• v.43 no.2
• /
• pp.164-168
• /
• 2015

Citric acid is produced via submerged fermentation using yeasts. Among eight different strains of yeast, Candida zeylanoides was chosen as the strain for producing citric acid and optimized for various C/N ratios and effects of phosphate or Fe²⁺ ions in a clean carbon source medium (glucose: fructose, 1:1). The yield of citric acid was maximized at a C/N ratio of 40/1, a phosphate addition of 1.0 g/l, and an Fe²⁺ ion concentration of less than 50 mg/l, yielding up to 91 g/L in the broth with 18.5 g/l of isocitric acid in a six-day fermentation period using a pre-treated molasses medium. The yield of batch culture was 0.51 (Y_p/s, g/g) in a 5 L-Jar fermenter.

• KSBB Journal
• /
• v.20 no.1 s.90
• /
• pp.60-65
• /
• 2005

The objective of this work was to evaluate a solid-state fermentation process for the practical production of arachidonic acid(AA) by Mortierella alpina ATCC 32222. In the present investigation, batch culture kinetics for both submerged- and solid-state fermentations was carried out at $25^{\circ}C$ to identify the relationship between growth and arachidonic acid (AA) production. Glucose and yeast extract were used in submerged fermentations by using flasks, while rice bran was used as a sole raw material in the other type of fermentations by using a series of Petri dishes. It was evident that a mixed-growth associated pattern existed between the two variables, irrespective of modes of fermentations. The effect of carbon to nitrogen (CfN) ratio on AA production in solid-state fermentation was studied in the range of 6.5 - 20. As a result, an optimum condition was found to be 6.5. Supplementary carbon source was not necessary to meet the optimum C/N ratio. Unlike the Previous results obtained by other researchers, a supplement of sodium glutamate up to $4\%$ (w/w) to the rice bran medium did not have a positive effect on the AA productivity. However, an increase in AA productivity was obtained with the rice bran medium supplemented with sesame oil.

• KSBB Journal
• /
• v.11 no.1
• /
• pp.65-70
• /
• 1996

Sodium gluconate was produced by neutralization of gluconic acid formed during the submerged culture fermentation of glucose with Aspergillus niger ACM 7. The fermentation characteristics of Aspergillus niger ACM 7 were investigated quantitatively according to the change of the initial glucose concentrations and the initial pHs of fermentation broth. The maximum specific growth rate was estimated to be $0.20hr^{-1}$ at 95g/$\ell$ of initial glucose concentration. The maximum fermentability of sodium gluconate was 95% at the initial glucose concentration of 26g/$\ell$. However, the maximum sodium gluconate productivity was 1.18g/$\ell$/hr when the initial glucose concentration was 110g/$\ell$. The optimum pH was found to be 5.5 for both the cell growth and the sodium gluconate production. With optimized culture conditions, the productivity of sodium gluconate in a fed-batch culture(production fermentor, 16,000$\ell$) increased up to 7.1g/$\ell$/hr.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.325-328
• /
• 2001

Teicoplanin is a glycopeptide antibiotic produced by Actinoplanes teichomyceticus novo sp. A TCC 31121. It is active against Gram-positive bacteria and it is under evaluation for use in man. Teicoplanin mixture in fermentation broth contains major amounts of teicoplanin $A_1$ and $A_2$ and a minor amount tcicoplanin of $A_3$. Commercial teicoplanin product is composed of five major components of very similar polarity, designated T-$A_2$-l, 2, 3, 4 and 5, and the more polor component, designated T -$A_3$. The culture conditions were studied in order that hydrophilic teicoplanin $A_2$ components are more produced but hydrophobic teicoplanin $A_1$ with lower bioactivity are less produced in submerged culture. Effects of culture pH and nutrients on the biosynthes ratio of teicoplanin $A_1$ and $A_2$ were confirmed in flask culture using MOPS buffer system through TLC, bioautography and bioassay. It was elucidated that optimal pH is 7.4 for teicoplanin $A_2$ biosynthesis but is 5.2 for teicoplanin $A_1$ biosynthesis, and that trace elements stimulate $A_2$ production but malt extract stimulate $A_1$production.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.2
• /
• pp.245-253
• /
• 2014

A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDS-PAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the ${\alpha}$ and ${\beta}$ chains of fibrinogen followed by the ${\gamma}$ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at $45^{\circ}C$ and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.4
• /
• pp.587-595
• /
• 2019

Pharmacological research on (CHA), a marine-derived quinazolinone alkaloid with significant cytotoxic activity, is restricted by low yields and is a problem that needs to be settled urgently. In this work, the selection of additional nitrogen sources and the optimization of additional concentrations and longer fermentation times using ammonium acetate, were investigated. CHA production was optimized to 62.1 mg/l with the addition of 50 mM ammonium acetate at 120 h of the fermentation in the shaker flask. This feeding strategy significantly increased 3-deoxy-arabino-heptulosonate-7-phosphate synthase activity and transcript levels of critical genes (laeA, dahp, and trpC) in the shikimate pathway compared with the non-treatment group. In addition, the selection of the feeding rate (0.01 and $0.03g/l{\cdot}h$) was investigated in a 5-L bioreactor. As a result, CHA production was increased by 57.9 mg/l with a $0.01g/l{\cdot}h$ ammonium acetate feeding rate. This work shows that the strategy of ammonium acetate supplementation had an effective role in improving CHA production by Aspergillus fumigatus CY018. It also shows that this strategy could serve as an important example of large-scale fermentation of a marine fungus in submerged culture.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.11
• /
• pp.1819-1826
• /
• 2015

Poly(ε-ʟ-lysine) (ε-PL) is a novel bioactive polymer secreted by filamentous bacteria. Owing to lack of a genetic system for most ε-PL-producing strains, very little research on enhancing ε-PL biosynthesis by genetic manipulation has been reported. In this study, an effective genetic system was established via intergeneric conjugal transfer for Streptomyces albulus PD-1, a famous ε-PL-producing strain. Using the established genetic system, the Vitreoscilla hemoglobin (VHb) gene was integrated into the chromosome of S. albulus PD-1 to alleviate oxygen limitation and to enhance the biosynthesis of ε-PL in submerged fermentation. Ultimately, the production of ε-PL increased from 22.7 g/l to 34.2 g/l after fed-batch culture in a 5 L bioreactor. Determination of the oxygen uptake rate, transcriptional level of ε-PL synthetase gene, and ATP level unveiled that the expression of VHb in S. albulus PD-1 enhanced ε-PL biosynthesis by improving respiration and ATP supply. To the best of our knowledge, this is the first report on enhancing ε-PL production by chromosomal integration of the VHb gene in an ε-PL-producing strain, and it will open a new avenue for ε-PL production.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.1
• /
• pp.57-65
• /
• 2015

β-Glucosidase production by the white rot fungus Flammulina velutipes CFK 3111 was evaluated using different carbon and nitrogen sources under submerged fermentation. Maximal extracellular enzyme production was 1.6 U/ml, corresponding to a culture grown in sucrose 40 g/land asparagine 10 g/l. High production yield was also obtained with glucose 10 g/land asparagine 4 g/l medium (0.5 U/ml). Parameters affecting the enzyme activity were studied using p-nitrophenyl-β-_D-glucopyranoside as the substrate. Optimal activity was found at 50℃ and pHs 5.0 to 6.0. Under these conditions, β-glucosidase retained 25% of its initial activity after 12 h of incubation and exhibited a half-life of 5 h. The addition of MgCl₂, urea, and ethanol enhanced the β-glucosidase activity up to 47%, whereas FeCl₂, CuSO₄, Cd(NO₃)₂, and cetyltrimethylammonium bromide inflicted a strong inhibitory effect. Glucose and cellobiose also showed an inhibitory effect on the β-glucosidase activity in a concentration-dependent manner. The enzyme had an estimated molecular mass of 75 kDa. To the best of our knowledge, F. velutipes CFK 3111 β-glucosidase production is amongst the highest reported to date, in a basidiomycetous fungus.