• Title/Summary/Keyword: optimal medium constituent

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Identification of characterization and statistical optimization of medium constituent for Bacillus subtilis SCJ4 isolated from Korean traditional fermented food (전통 장류 유래 Bacillus subtilis SCJ4의 특성확인 및 통계학적 방법을 이용한 배양조건 최적화)

  • Jeong, Su-Ji;Yang, Hee-Jong;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.48-60
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    • 2015
  • 612 strains isolated from Korean traditional fermented food in Sunchang and their investigated biochemical characterization and ability of biogenic amines non-producing. We selected the SCJ4 having various activity by measurement of extracellular enzyme, antioxidant and antimicrobial activities. Selected strain SCJ4 by 16S rRNA sequencing and biochemical characterization was named Bacillus subtilis SCJ4. And then, we investigated cell growth of SCJ4, and optimized of culture medium constituents using response surface methodology as statistically method. Response surface methodology used Plackett-Burman experimental design for screening of medium constituent. Tryptone, peptone and $MgSO_4$ as medium constituent improving cell growth selected. In order to find out optimal concentration on each constituent, we carried out central composite design. Consequently, optimized concentrations of tryptone, peptone and $MgSO_4$ were predicted to be 15.35 g/L, 12.235 g/L, and 3.5 g/L respectively. Through the model verification, we confirmed about 1.28-fold improvement of the dried cell weight from 0.8767 g/L to 1.1222 g/L when compared to basal medium.

Effect of Medium Composition on Protopectinase Production from Bacillus subtilis EK11 (Bacillus subtilis EK11로부터 Protopectinase 생산을 위한 배지성분의 영향)

  • 이대희;박은경;문철환;하정욱;이승철;황용일
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.378-384
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    • 1999
  • Protopectinase (PPases) are heterologous group of enzymes that degrade pectin from the insoluble protopection which is constituent of the middle lamella and primary cell wall of higher plants by restricted depolymerization. From the previous report[6], enzymatically separated plant cells, which are produced from plant tissues by PPases treatment, showed well-conserved cellular components with their rigid cell wall and this characteristic is applicable to preparation of novel food material. The purpose of this study is to investigate the effect of medium composition of PPase production from Bacillus subtilis EK11 which was selected as a PPase producer. Various carbon sources and concentrations on PPase production were studied and corn starch at 0.7% was the most effective for production of PPase. Among the nitrogen sources, yeast extract was the most effective for PPase production and the effect of (NH4)2SO4 was notable as inotganic nitrogen source. Inorganic compounds such as KH2PO4, K2HPO4, Na3-citrate.2H2O and MgSO4 were optimized for PPase production. PPase activity was inhibited by the adition of Ba2+ or Zn2+. The optimal medium for PPase production was devised: 0.7% corn starch, 0.3% yeast extract, 1.4% KH2PO4, 0.6% K2HPO4, 0.1% Na3-citrate.2H2O and 0.02% MgSO4. PPase production by using the optimum medium was carried out with shaking cultivation at 37$^{\circ}C$. The maximum PPase activity of 256unit/ml could be obtained after the cultivation for 48hrs. The activity was increased about 2.2timesthan the activity, 112 unit/ml, in basal medium.

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Statistical Optimization of Production Medium for Enhanced Production of Succinic Acid Produced by Anaerobic Fermentations of Actinobacillus succinogenes (Actinobacillus succinogenes의 혐기성배양에 의해 생합성 되는 숙신산의 생산성 향상을 위한 통계적 생산배지 최적화)

  • Park, Sang-Min;Chun, Gie-Taek
    • KSBB Journal
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    • v.29 no.3
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    • pp.165-178
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    • 2014
  • Statistical medium optimization has been carried out for the production of succinic acid in anaerobic fermentations of Actinobacillus succinogenes. Succinic acid utilized as a precursor of many industrially important chemicals is a fourcarbon dicarboxylic acid, biosynthesized as one of the fermentation products of anaerobic metabolism by A. succinogenes. Through OFAT (one factor at a time) experiments, corn steep liquor (CSL), a very cheap agricultural byproduct, was found to have significant effects on enhanced production of succinic acid, when supplemented along with yeast extract. Hence, using these factors including glucose as a carbon/energy source, interactive effects were investigated through $2^n$ full factorial design (FFD) experiments, showing that the concentration of each component (i.e., glucose, yeast extract and CSL) should be higher. Further statistical experiments were conducted along the steepest ascent path, followed by response surface method (RSM) in order to find out optimal concentrations of each constituent. Consequently, optimized concentrations of glucose, yeast extract and CSL were observed to be 180 g/L, 15.08 g/L and 20.75 g/L respectively (10 g/L of $NaHCO_3$ and 100 g/L of $MgCO_3$ to be supplemented as bicarbonate suppliers), with the estimated production level of succinic acid to be 92.9 g/L (about 3.5 fold higher productivity as compared to the initial medium). Notably, the RSM-estimated production level was almost similar to the amount of succinic acid (92.9 g/L vs. 89.1 g/L) produced through the actual fermentation process performed using the statistically optimized production medium.

Optimal Condition for Mycelial Growth of Beauveria bassiana and Its Extracellular Enzyme Activity (백강균(Beauveria bassiana)의 균사체 최적 배양조건 및 효소활성)

  • 민응기;한영환
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.50-53
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    • 2002
  • The optimum temperature and pH for mycelial growth of B. bassiana DGUM 34001 were $24^{\circ}C$ and pH 7.0, respectively. Among the complex media used, mushroom complex medium (MCM) was the most favorable for mycelial growth. When Czapek-Dox medium was used as a minimal medium, glucose was an excellent source for carbon and energy. Soytone and sodium phosphate were favorable constituent for culture medium as a source of organic nitrogen and phosphorus, respectively. When the fungus was grown in MCM broth, the specific activity of extracellular enzyme of ${\alpha}$-amylase, lipase, chitinase, CMCase and pretease were 297.0, 0.058, 0.33, 0.21 and 22.8 units/mg protein, respectively. When various sources of organic nitrogen and chitin were supplemented to determine the production of enzymes, casein and soluble chitosan enhanced the production of extracellular protease and chitinase.

Statistical Optimization of Production Medium for Enhanced Production of Itaconic Acid Biosynthesized by Fungal Cells of Aspergillus terreus (Aspergillus terreus에 의해 생합성되는 이타콘산의 생산성 증가를 위한 통계적 생산배지 최적화)

  • Jang, Yong-Man;Shin, Woo-Shik;Lee, Do-Hoon;Kim, Sang-Yong;Park, Chul-Hwan;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.24 no.1
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    • pp.30-40
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    • 2009
  • Statistical optimization of the production medium was carried out in order to find an optimal medium composition in itaconic acid fermentation process. Itaconic acid utilized in the manufacture of various synthetic resins is a dicarboxylic acid biosynthesized by fungal cells of Aspergillus terreus in a branch of the TCA cycle via decarboxylation of cis-aconitate. Through OFAT (one factor at a time) experiments, six components (glucose, fructose, sucrose, soluble starch, soybean meal and cottonseed flour) were found to have significant effects on itaconic production among various carbon- and nitrogen-sources. Hence, using these six factors, interactive effects were investigated via fractional factorial design, showing that the initial concentrations of sucrose and cottonseed flour should be high for enhanced production of itaconic acid. Furthermore, through full factorial design (FFD) experiments, negative effects of $KH_2PO_4$ and $MgSO_4$ on itaconic acid biosynthesis were demonstrated, when excess amounts of the each component were initially added. Based on the FFD analysis, further statistical experiments were conducted along the steepest ascent path, followed by response surface method (RSM) in order to obtain optimal concentrations of the constituent nutrients. As a result, optimized concentrations of sucrose and cottonseed flour were found to be 90.4g/L and 53.8g/L respectively, with the corresponding production level of itaconic acid to be 4.36 g/L (about 7 fold higher productivity as compared to the previous production medium). From these experimental results, it was assumed that optimum ratio of the constituent carbon (sucrose) and nitrogen (cottonseed flour) sources was one of the most important factors for the enhanced production of itaconic acid.

Isolation and Characterization of Pectinase-Producing Bacillus sp. BS-214 (Pectinase를 생산하는 Bacillus sp. BS-214의 분리 및 특성)

  • 전병삼;차재영;송재영;이강덕;김범규;이영춘
    • Journal of Life Science
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    • v.10 no.1
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    • pp.101-106
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    • 2000
  • A bacterial strain BS-214 producing extracellular pectinase was isolated from soil. The isolated bacterium was identified as a strain of Bacillus so. based on the morphological, biochemical, and physiological characteristics. Cell growth and pectinase activity of Bacillus sp. BS-214 were reached to a mixium in the culture condition of pH 8.5 at 4$0^{\circ}C$. Production of pectinase by the strain was the highest when polygalacturonic acid was added to culture medium as a carbon source, and its optimal concentration was 1%. Also, yeast extract was used as the best nitrogen source for the production of pectinase by the concentration of 0.25%. Decomposition of a constituent of Edzeworthia papyrifera by the strain was observed by scanning electron microscope.

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Screening of Non-Biogenic-Amine-Producing Bacillus subtilis and Medium Optimization for Improving Biomass by the Response Surface Methodology (바이오제닉 아민 비생성 Bacillus subtilis의 선별 및 반응표면 분석법에 의한 균체량 증가를 위한 배지 최적화)

  • Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Seong-Yeop;Heo, Ju-Hee;Choi, Nack-Shick;Jeong, Do-Youn
    • Journal of Life Science
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    • v.26 no.5
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    • pp.571-583
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    • 2016
  • Biogenic amines are produced primarily by microorganisms found in fermented foods and are often implicated in poisoning incidents in humans. In this study, 620 strains of microorganisms were isolated from traditional Korean fermented food in Sunchang in order to screen for non-biogenicamine-producing microorganisms present in these foods. One strain was identified and named Bacillus subtilis SCJ1, by using 16S rRNA sequencing and biochemical characterization. We investigated the cell growth of this organism in order to understand its potential for industrial application. To this end, we optimized the culture medium constituents by using the response surface methodology. The Plackett-Burman experimental design was used for screening of the medium constituents, such as molasses, yeast extract and peptone, for improving cell growth. In order to determine the optimal concentration of each constituent, we used a central composite design. Consequently, the optimized concentrations of molasses, yeast extract and peptone were predicted to be 27.5 g/l, 7.5 g/l and 17.5 g/l, respectively. By model verification, we confirmed that a 1.49-fold increase in dry cell weight compared to the basal medium-from 1.32 g/l, to 1.9722 g/l-was achieved.

Statistical Optimization of Culture Conditions of Probiotic Lactobacillus brevis SBB07 for Enhanced Cell Growth (프로바이오틱 Lactobacillus brevis SBB07의 균체량 증가를 위한 배양 조건 최적화)

  • Jeong, Su-Ji;Yang, Hee-Jong;Ryu, Myeong Seon;Seo, Ji Won;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Journal of Life Science
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    • v.28 no.5
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    • pp.577-586
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    • 2018
  • We recently reported the potential probiotic properties of Lactobacillus brevis SBB07 isolated from blueberries. The present study investigates the effect of culture conditions such as temperature, initial pH, culture time, and medium constituent for industrial application. The ingredients of the medium to improve cell growth were selected by Plackett-Burman design (PBD) and central composite design (CCD) within a desirable range. The PBD was applied with 19 factors: seven carbon sources, six nitrogen sources, and six microelements. Protease peptone, corn steep powder (CSP), and yeast extract were found to be significant factors for the growth of SBB07. The CCD was then applied with three variables found from the PBD at five levels, and the optimum values were decided for the three variables: protease peptone, CSP, and yeast extract. In the case of the growth of SBB07, the proposed optimal media contained 2.0% protease peptone, 2.5% CSP, and 2.0% yeast extract, and the maximum dried-cell weight was predicted to be 2.93963 g/l. By the model verification, it was confirmed that the predicted and actual results are similar. Finally, the study investigated the effects of incubation temperature and initial pH at the optimized medium. It was confirmed that the dried-cell weight increased from $2.2933{\pm}0.0601g/l$ to $3.85{\pm}0.0265g/l$ when compared to the basal medium at $37^{\circ}C$ and initial pH 8.0. Establishing the optimal culture condition for SBB07 provides good potential for applications in probiotics and can serve as the foundation for the industrialization of materials.

Optimization of Medium for the Carotenoid Production by Rhodobacter sphaeroides PS-24 Using Response Surface Methodology (반응 표면 분석법을 사용한 Rhodobacter sphaeroides PS-24 유래 carotenoid 생산 배지 최적화)

  • Bong, Ki-Moon;Kim, Kong-Min;Seo, Min-Kyoung;Han, Ji-Hee;Park, In-Chul;Lee, Chul-Won;Kim, Pyoung-Il
    • Korean Journal of Organic Agriculture
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    • v.25 no.1
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    • pp.135-148
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    • 2017
  • Response Surface Methodology (RSM), which is combining with Plackett-Burman design and Box-Behnken experimental design, was applied to optimize the ratios of the nutrient components for carotenoid production by Rhodobacter sphaeroides PS-24 in liquid state fermentation. Nine nutrient ingredients containing yeast extract, sodium acetate, NaCl, $K_2HPO_4$, $MgSO_4$, mono-sodium glutamate, $Na_2CO_3$, $NH_4Cl$ and $CaCl_2$ were finally selected for optimizing the medium composition based on their statistical significance and positive effects on carotenoid yield. Box-Behnken design was employed for further optimization of the selected nutrient components in order to increase carotenoid production. Based on the Box-Behnken assay data, the secondary order coefficient model was set up to investigate the relationship between the carotenoid productivity and nutrient ingredients. The important factors having influence on optimal medium constituents for carotenoid production by Rhodobacter sphaeroides PS-24 were determined as follows: yeast extract 1.23 g, sodium acetate 1 g, $NH_4Cl$ 1.75 g, NaCl 2.5 g, $K_2HPO_4$ 2 g, $MgSO_4$ 1.0 g, mono-sodium glutamate 7.5 g, $Na_2CO_3$ 3.71 g, $NH_4Cl$ 3.5g, $CaCl_2$ 0.01 g, per liter. Maximum carotenoid yield of 18.11 mg/L was measured by confirmatory experiment in liquid culture using 500 L fermenter.

An Ethanol Extract of the Brown Seaweed Hizikia fusiformis and Its Active Constituent, Fucosterol, Extend the Lifespan of the Nematode Caenorhabditis elegans (갈조류 톳(Hizikia fusiformis)의 에탄올추출물 및 이의 활성성분 fucosterol에 의한 예쁜꼬마 선충의 수명 연장)

  • Oktaviani, Diyah Fatimah;Bae, Young-Seuk;Meinita, Maria Dyah Nur;Moon, Il Soo;Hong, Yong-Ki
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1120-1125
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    • 2019
  • The short-lived nematode Caenorhabditis elegans has been used as a model organism for many studies, including lifespan extension. To screen common seaweeds for natural anti-aging agents, the lifespan of C. elegans (N2 wild-type strain) was measured by its hatch rate, growth rate, survival rate, chemotaxis, brood size, and egg-laying time after exposure to nematode growth medium (NGM) containing seaweed extracts. Approximately 30 animals synchronized at the first larval stage were incubated until they reached their adult stages before laying their eggs and were transferred to fresh NGM every 3 days. We also identified the major active compound from the seaweed by gas chromatography-mass spectrometry and tested its optimal dose for longevity. Of 13 common seaweed species, an ethanol extract of the brown seaweed Hizikia fusiformis showed the greatest effect on hatching, growth, and survival rates. The lifespan of C. elegans was significantly expanded 1.54-fold and 1.23-fold in the presence of the ethanol extract (0.05 mg/ml) and the main active component, fucosterol (0.05 mg/ml), respectively. Exposure to the ethanol extract also increased chemotaxis 1.13-fold, decreased brood size 0.74-fold, and shortened egg-laying time 0.96-fold. These results suggest that the aquaculturable H. fusiformis may be a promising source of a diet supplement to support health care.