• 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.

• Microbiology and Biotechnology Letters
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• v.47 no.2
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• pp.220-233
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• 2019

Bacterial lysates have become a common ingredient for natural health care. Lactic acid bacteria (LAB) could serve as potential candidates for lysate production: the lactic acids produced by LAB have been utilized for their moisturizing, antimicrobial, and rejuvenating effects, while other substances provide topical benefits and health effects for the skin. Our study aimed to obtain lysate from a LAB S. macedonicus MBF 10-2 through an optimized fermentation using the Response Surface Methodology. Strain MBF10-2 was cultivated in a 2L fermenter tank in de Man Rogosa and Sharpe (MRS) medium and in plant-based peptone modified MRS, i.e. Soy-peptone and Vegitone. The duration and the medium composition (dextrose and soy peptone or proteose peptone) were adjusted to obtain an optimum production of cell lysate. Central Composite Design was employed for Design Expert 7.0.0 by adjusting 3 factors: dextrose (1%, 1.5%, 2%, 2.5%, 3%), soy or proteose peptone (0.5%, 0.75%, 1%, 1.25% and 1.5%), and duration of fermentation (8, 10, 12 14, and 16 h for MRS-Soy peptone and 15, 17, 19, 21, and 23 h for MRS Vegitone). Bacteriocin-Like Inhibitor Substance activity of lysate and pH were used as indicators. The optimum condition for lysate production using MRS Soy Peptone and Vegitone are as follows: dextrose concentration 2.5%, plant-based peptone 1.25%, while optimum fermentation duration were 11.18 h (MRS Soy Peptone) and 17 h (MRS Vegitone) with a starter concentration of 10% at $OD_{600nm}$ $0.2{\pm}0.05$. However, the standard MRS medium produced better quality lysate compared to MRS plant-based peptones.

• The Korean Journal of Mycology
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• v.47 no.4
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• pp.359-371
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• 2019

To optimize the expression and secretion of ferritin protein associated with ion storage in the mushroom, Pleurotus eryngii, a recombinant secretion vector, harboring the ferritin gene, was constructed using a pPEVPR1b vector under the control of the CaMV 35S promoter and signal sequence of pathogen related protein (PR1b). The ferritin gene was isolated from the T-Fer vector following digestion with EcoRI and HindIII. The gene was then introduced into the pPEVPR1b secretion vector, and it was then named pPEVPR1b-Fer. The recombinant vector was transferred into P. eryngii via Agrobacterium tumefaciens-mediated transformation. The transformants were selected on MCM medium supplemented with kanamycin and its expression was confirmed by SDS-PAGE and western blotting. Expression of ferritin protein was optimized by modifying the culture conditions such as incubation time and temperature in batch and 20 L airlift type fermenter. The optimal conditions for ferritin production were achieved at 25℃ and after incubating for 8 days on MCM medium. The amount of ferritin protein was 2.4 mg/g mycelia, as measured by a quantitative protein assay. However, the signal sequence of PR1b (32 amino acids) seems to be correctly processed by peptidase and ferritin protein may be targeted in the apoplast region of mycelia, and it might not be secreted in the culture medium. The iron binding activity was confirmed by Perls' staining in a 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in P. eryngii mycelia. Mycelium powder containing ferritin was tested as a feed additive in broilers. The addition of ferritin powder stimulated the growth of young broilers and improved their feed efficiency and production index.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.3
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• pp.376-380
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• 2006

An experiment was conducted to study the effect of temperature and pH on in vitro nutrient degradability, volatile fatty acid profile and methane production. The fermenter used was the semi-continuous system, known as the rumen simulation technique (RUSITEC). Sixteen cylinders were used at one time with a volume of 800 ml, the dilution rate was set at 3.5%/hour, the infused buffer being McDougall's artificial saliva. Basal diet (9.6 g DM) used in RUSITEC consisted of (DM) 6.40 g Timothy hay, 1.86 g crushed corn and 1.34 g soybean meal. The food for the fermentation vessel was provided in nylon bags, which were gently agitated in the liquid phase. The experiment lasted for 17 d with all the samples taken during the last 5 d. Treatments were allocated at random to four vessels each and were (1) two temperature levels of $39^{\circ}C$ and $41^{\circ}C$ (2) two pH levels of 6.0 and 7.0. The total diet contained ($g\;kg^{-1}$ DM) 957 OM, 115 CP and $167MJ\;kg^{-1}$ (DM) GE. Although increase in temperature from $39^{\circ}C$ to $41^{\circ}C$ reduced degradation of major nutrients in vitro, it was non-significant. Interaction effect of temperature with pH also reflected a similar trend. However, pH showed a significant (p<0.05) negative effect on the degradability of all the nutrients in vitro. Altering the in vitro pH from 7 to 6 caused marked reduction in DMD from 60.2 to 41.8, CPD from 76.3 to 55.3 and GED from 55.3 to 35.1, respectively. Low pH (6) depressed total VFA production (61.9 vs. 34.9 mM) as well as acetate to propionate ratio in vitro (from 2.0 to 1.5) when compared to pH 7. Compared to pH 7, total gas production decreased from 1,841 ml to 1,148 ml at pH 6, $CO_2$ and $CH_4$ production also reduced from 639 to 260 ml and 138 to 45 ml, respectively. This study supported the premise that pH is one of the principal factors affecting the microbial production of volatile fatty acids and gas. Regulating the ruminal pH to increase bacterial activity may be one of the methods to optimize VFA production, reduce methane and, possibly, improve animal performance.

• Journal of Life Science
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• v.18 no.7
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• pp.980-985
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• 2008

This study was carried out to develope an alcoholic drink by fermentation of onion extract using Saccharomyces cerevisiae. The optimal conditions for ethanol production were obtained by standing culture at $25^{\circ}C$ for 5 days with 5% inoculum volume. At the results by flask culture, the growth curve of used S. cerevisiae reached to the stantionary phase at 48 hr and the death phase at 90 hr, whereas ethanol production reached maximum at 114 hr. Under the above conditions, a large scale production was carried out. A standing culture in 5 l fermenter showed the similar results to its flask culture, but progressed 24 hr rapidly more than that of the flask culture. A fed-batch culture was performed by addition of the onionic medium supplemented with 10% (v/v) sucrose after 72 hr from the fermenting start. The fed-batch culture could prevent S. cerevisiae from entering into the death phase and maintain constant level of alcohol production. A continuous culture was able to carry out by adding per every 24 hr the onionic medium supplemented with 10% (v/v) sucrose after 72 hr from the fermenting start. Although S. cerevisiae used showed a little decreased growth, alcohol production maintained roughly the constant level at the maximum yield. To enhance the quality of this alcoholic drink, $2-O-{\alpha}-D-glucopyranosyl$ L-ascorbic acid (AA-2G) was supplemented into the onion extract of the substrate for fermentation. As resulted at this study, this alcoholic drink containing AA-2G should be used as a functional fermented alcohol drink strengthened with vitamin C.

• Journal of Life Science
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• v.32 no.7
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• pp.510-522
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• 2022

In this study, to biosynthesize PHA with properties more similar to polypropylene, a Bacillus sp. EMK-5020 strain that biosynthesized poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was isolated from soil. Bacillus sp. EMK-5020 strain biosynthesized PHBV containing 1.3% 3-hydroxyvalerate (3HV) using reducing sugar contained in Makgeolli lees enzymatic hydrolysate (MLEH) as a single carbon source. As the amount of propionic acid, which was added as a second carbon source, increased, the content of 3HV also increased. PHBV containing up to 48.6% of 3HV was synthesized when 1.0 g/l of propionic acid was added. Based on these results, the strain was cultured for 72 hr in a 3 l fermenter using reducing sugar in MLEH (20 g/l) and propionic acid (1 g/l) as the main and secondary carbon sources, respectively. As a result, 6.4 g/l DCW and 50 wt% of PHBV (MLEH-PHBV) containing 8.9% 3HV were biosynthesized. Through gel permeation chromatography and thermogravimetric analysis, it was confirmed that the average molecular weight and the decomposition temperature of MLEH-PHBV were 152 kDa and 273℃, respectively. In conclusion, the Bacillus sp. EMK-5020 strain could biosynthesize PHBV containing various 3HV fractions when MLEH and propionic acid were used as carbon sources, and PHBV-MLEH containing 8.9% 3HV was confirmed to have higher thermal stability than standard PHBV (8% 3HV).

• The Korean Journal of Food And Nutrition
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• v.29 no.6
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• pp.962-970
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• 2016

Commercialized production of onion vinegar, which has biological activities formed through alcohol and acetic acid fermentation, requires standardization. The objective of this study was to determine optimal conditions of sugar contents ($11{\sim}15^{\circ}Brix$) and agitation rate (100~300 rpm) of fermenter in the alcohol-acetic fermentation for producing onion vinegar. The alcohol and total acidity contents increased, whereas contents of total sugars decreased during alcohol fermentation. Contents of alcohol of 13 and $15^{\circ}Brix$ reactants were about 8% in 36 hr and total acidities of all samples were below 0.2% in 60 hr. During acetic fermentation, total acidity increased with highest value at 9 days (3.2% in 100 rpm), 10 days (4.1% in 200 rpm) and 8 days (4.3% in 300 rpm), respectively. From these results, sugar contents ($13^{\circ}Brix$) were measured for alcohol fermentation and agitation rate (300 rpm) for fast fermentation method of vinegar. The contents of total phenols, flavonoids and quercetin in onion vinegar were 33.3 mg/100 g, 3.0 mg/100 g and 2.0 mg/100 g, respectively. Onion vinegar showed an antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium, Escherichia coli and Enterobacter aerogenes. Antioxidant effect of onion vinegar was 26.23% in DPPH radical inhibition and 58.58% in superoxide dismutase like activity, respectively. Fibrinolytic activity was 1.51 plasmin unit/mL in onion vinegar. In conclusion, onion vinegar processed by alcohol and acetic fermentation had nutritional values and potential biological activities.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.1
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• pp.85-92
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• 2005

The performance of a novel fermentation process, adopting a sludge blanket type configuration for higher hydrolysis/acidogenesis of the municipal primary sludge, was investigated under batch and semi-continuous conditions with various pH and temperature conditions. This acid elutriation slurry reactor provided higher system performance with a short HRT (5 days) and higher acidogenic effluent quality under pH 9 and thermophilic ($55^{\circ}C$) conditions. The hydrolysis of the sludge was revealed to be significantly dependent on seasonal effects for sludge characteristics but with little impact on acidogenesis. Based on the rainy season at the optimum conditions, VFA production and recovery fraction ($VFA_{COD}/COD$) were $0.18\;g\;VFA_{COD}\;g^{-1}\;VSS_{COD}$ and 63%. As byproducts, nitrogen and phosphorus releasing were $0.006\;g\;N\;g^{-1}\;VSS_{COD}$ and $0.003\;g\;P\;g^{-1}\;VSS_{COD}$, respectively. For the mass balance in a full-scale plant($Q=158,880\;m^3\;day^{-1}$) based on the rainy season, the VFA and non-VFA(as COD) production were $3,110\;kg\;VFA_{COD}\;day^{-1}$ and $1,800\;kg\;COD\;day^{-1}$, resulting in an increase of organics of $31\;mg\;COD\;L^{-1}$ and $20\;mg\;VFA_{COD}\;L^{-1}$ and nutrients of $0.7\;mg\;N\;L^{-1}$ and $0.3\;mg\;P\;L^{-1}$ in the influent sewage. The economical benefit from this process application was estimated to be about $67 per $1,000m^3$ of sewage except for energy requirements and also, better benefits can be expected during the dry season. Also, the results revealed that the process has various additional advantages such as pathogen-free stabilized solids production, excellent solids control and economical benefits.

• Food Science of Animal Resources
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• v.29 no.4
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• pp.437-444
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• 2009

${\gamma}-Aminobutyric$ acid (GABA) producing lactic acid bacteria, Lactobacillus acidophilus RMK567 was cultivated in 50 L of sterilized MRS broth using a fermenter at $40^{\circ}C$ for 24 h. The cell number was increased to $10.04{\pm}0.13$ Log CFU/mL with a growth rate constant (k) of 0.454 generation/h and a generation time (g) of 2.303 h after a lapse of a lag phase (L) of 5.16 h. A total of 487 g of cell paste with 40.5% moisture was harvested with viable cell number of 12.48 Log CFU/g cell paste. The cell pastes after preparation with glycerol, glucose, and polydextrose as cryo-protectants were lyophilized under a vacuum of 84 m torr. A total of 408 g of freeze dried (FD) cell powders were mixed with a commercial strain of Streptococcus thermophilus to prepare of three types FD starter cultures with the viable cell numbers of 12.42 (FDA-GY), 12.60 (FDBGG) and 12.91 (FDC-GP) Log CFU/g. During preservation the FD cultures at -$18^{\circ}C$, the cell viability of the FD starter cultures were rapidly dropped to below 3.24% of the day of storage. No significant difference was found in the cell viabilities among three types of FD starters cultures, but significant difference (p<0.01) was found in storage periods. Yoghurts fermented through FD starter culture of L. acidophilus RMK567 were determined to contain $155.16{\pm}8.53$ ppm, $243.82{\pm}4.27$ ppm, and $198.64{\pm}23.46$ ppm of GABA, respectively. This study shows that GABA production activity of L. acidophilus RMK567 is not affected during the freeze drying process and would be available for commercial production of yoghurt containing high GABA content.

• Korean Journal of Food Science and Technology
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• v.31 no.4
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• pp.1035-1043
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• 1999

Bacteriocins are natural antimicrobial compounds produced by many microorganisms associated with foods, so that there is currently much interest in their use as food biopreservatives. Goal of this study was to partially evaluate lacticin NK24 as a food biopreservative by showing antimicrobial activity of L. lactis NK24 and lacticin NK24 against food-borne spoilage and pathogenic bacteria, respectively. Lactic acid bacteria NK24 isolated from jeot-gal, Korean fermented fish foods, was tentatively identified as Lactococcus lactis and showed broad spectrum of activity against all of spoilage and pathogenic bacteria tested by deferred method. Bacteriocin production in jar fermenter was detected at the mid-log growth phase, and reached the maximum at the early stationary phase, but decreased after the stationary phase. Lacticin NK24 was partially purified by 75% ammonium sulfate precipitation followed by subsequent dialysis. This partially purified lacticin NK24 showed antimicrobial activity against Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Clostridium perfringens, some bacilli, Listeria monocytogenes, Listeria ivanovii, Sphin-gomonas pausimobilis, Escherichia coli and Pseudomonas aeruginosa. Thus, lacticin NK24 examined in this study show promise as a biopreservative be-cause of their broad spectrum of activity.