• Title/Summary/Keyword: statistical medium optimization

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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.

Screening of Biogenic Amine Non-Producing Yeast and Optimization of Culture Conditions Using Statistical Method for Manufacturing Black Raspberry Wine (복분자 와인 제조를 위한 바이오제닉 아민 비생성 효모의 선별 및 통계학적 기법을 이용한 배양조건 최적화)

  • Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Seong-Yeop;Heo, Ju-Hee;Jeong, Do-Youn
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.4
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    • pp.592-601
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    • 2015
  • Rubus coreanus is known as Korean black raspberry, native to Korea, Japan, and China. Preliminary studies evaluating their potential for cancer treatment in mammalian test systems are ongoing. In recent years, interest has been renewed due to their high levels of anthocyanins. Anthocyanins in black raspberry are important due to their potential health benefits as dietary antioxidant, anti-inflammatory compound, and as a chemopreventive agent. In the present study, Saccharomyces cerevisiae BA29 was isolated from black raspberry fruit and fruit juice as a biogenic amine non-producing strain for manufacturing of black raspberry wine, after which we investigated its characteristics: biogenic amine-producing ability, cell growth ability, alcohol-fermentation ability, and resistance to alcohol, glucose, and sulfur dioxide. Based on preliminary experiments, we optimized culture medium compositions for improving dried cell weight of S. cerevisiae BA29 by response surface methodology (RSM) as a statistical method. Design for RSM used a central composite design, and molasses with the industrial applicability was used as a carbon source. Through statistical analysis, we obtained optimum values as follows: molasses 200 g/L, peptone 30 g/L, and yeast extract 40 g/L. For the model verification, we confirmed about 3-fold improvement of dried cell weight from 6.39 to 20.9167 g/L compared to basal yeast peptone dextrose medium. Finally, we manufactured black raspberry wine using S. cerevisiae BA29 and produced alcohol of 20.33%. In conclusion, S. cerevisiae isolated from black raspberry fruit and juices has a great potential in the fermentation of black raspberry wine.

Statistical Optimization for Improved Production of Cyclosporin A in Solid-State Fermentation

  • Survase, Shrikant A.;Annapure, Uday S.;Singhal, Rekha S.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1385-1392
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    • 2009
  • This work evaluates the effect of different amino acids on production of Cyclosporin (CyA) production in solid-state fermentation that was previously optimized for different fermentation parameters by one factor at-a-time for the maximum production of CyA by Tolypocladium inflatum MTCC557. Based on the Plackett-Burman design, glycerol, ammonium sulfate, $FeCl_3$, and inoculum size were selected for further optimization by response surface methodology (RSM). After identifying effective nutrients, RSM was used to develop mathematical model equations, study responses, and establish the optimum concentrations of the key nutrients for higher CyA production. It was observed that supplementation of medium containing (% w/w) glycerol, 1.53; ammonium sulfate, 0.95; $FeCl_3$, 0.18; and inoculum size 6.4 ml/5g yielded a maximum of 7,106 mg/kg as compared with 6,480 mg CyA/kg substrate using one factor at-a-time. In the second step, the effect of amino acids on the production of CyA was studied. Addition of $_L$-valine and $_L$-leucine in combination after 20 h of fermentation resulted in maximum production of 8,166 mg/kg.

Coproduction of Thermostable Amylase and ${\beta}$-Galactosidase Enzymes by Geobacillus stearothermophilus SAB-40: Application of Plackett-Burman Design to Evaluate Culture Requirements Affecting Enzyme Production

  • Soliman, Nadia A.
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.695-703
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    • 2008
  • A locally isolated thermophile, Geobacillus sp. SAB-40, producing thermostable extracellular amylase constitutively and an induced intracellular ${\beta}$-galactosidase was characterized and identified based on 16S rRNA sequencing. A phylogenetic analysis then revealed its closeness to Geobacillus stearothermophilus. To evaluate the effect of the culture conditions on the coproduction of both enzymes by G stearothermophilus SAB-40, a Plackett-Burman fractional factorial design was applied to determine the impact of twenty variables. Among the tested variables, $CaCI_2$, the incubation time, $MgSO_4{\cdot}7H_2O$, and tryptone were found to be the most significant for encouraging amylase production. Lactose was found to promote ${\beta}$-galactosidase production, whereas starch had a significantly negative effect on lactase production. Based on a statistical analysis, a preoptimized medium attained the maximum production of amylase and ${\beta}$-galactosidase at 23.29 U/ml/ min and 12,958 U/mg biomass, respectively, which was 3-and 2-fold higher than the yield of amylase and lactase obtained with the basal medium, respectively.

Effects of Sucrose, Phosphate, and Calcium Carbonate on the Production of Pikromycin from Streptomyces venezuelae

  • Yi, Jeong Sang;Kim, Minsuk;Kim, Sung-Jin;Kim, Byung-Gee
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.496-502
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    • 2015
  • Polyketide secondary metabolites share common precursor pools, acyl-CoA. Thus, the effects of engineering strategies for heterologous and native secondary metabolite production are often determined by the measurement of pikromycin in Streptomyces venezuelae. It is hard to compare the effectiveness of engineering targets among published data owing to the different pikromycin production media used from one study to the other. To determine the most important nutritional factor and establish optimal culture conditions, medium optimization of pikromycin from Streptomyces venezuelae ATCC 15439 was studied with a statistical method, Plackett-Burman design. Nine variables (glucose, sucrose, peptone, (NH4)2SO4, K2HPO4, KH2PO4, NaCl, MgSO4·7H2O, and CaCO3) were analyzed for their effects on a response, pikromycin. Glucose, K2HPO4, and CaCO3 were determined to be the most significant factors. The path of the steepest ascent and response surface methodology about the three selected components were performed to study interactions among the three factors, and the fine-tune concentrations for maximized product yields. The significant variables and optimal concentrations were 139 g/1 sucrose, 5.29 g/l K2HPO4, and 0.081 g/l CaCO3, with the maximal pikromycin yield of 35.5 mg/l. Increases of the antibiotics production by 1.45-fold, 1.3-fold, and 1.98-fold, compared with unoptimized medium and two other pikromycin production media SCM and SGGP, respectively, were achieved.

Application of Factorial Experimental Designs for Optimization of Cyclosporin A Production by Tolypocladium inflatum in Submerged Culture

  • Abdel-Fattah, Y.R.;Enshasy, H. El;Anwar, M.;Omar, H.;Abolmagd, E.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1930-1936
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    • 2007
  • A sequential optimization strategy based on statistical experimental designs was employed to enhance the production of cyclosporin A (CyA) by Tolypocladium inflatum DSMZ 915 in a submerged culture. A 2-level Plackett-Burman design was used to screen the bioprocess parameters significantly influencing CyA production. Among the 11 variables tested, sucrose, ammonium sulfate, and soluble starch were selected, owing to their significant positive effect on CyA production. A response surface methodology (RSM) involving a 3-level Box-Behnken design was adopted to acquire the best process conditions. Thus, a polynomial model was created to correlate the relationship between the three variables and the CyA yield, and the optimal combination of the major media constituents for cyclosporin A production, evaluated using the nonlinear optimization algorithm of EXCEL-Solver, was as follows (g/l): sucrose, 20; starch, 20; and ammonium sulfate, 10. The predicted optimum CyA yield was 113 mg/l, which was 2-fold the amount obtained with the basal medium. Experimental verification of the predicted model resulted in a CyA yield of 110 mg/l, representing 97% of the theoretically calculated yield.

Application of Response Surface Methodology in Medium Optimization to Improve Lactic Acid Production by Lactobacillus paracasei SRCM201474 (반응표면분석법을 이용한 Lactobacillus paracasei SRCM201474의 생산배지 최적화)

  • Ha, Gwangsu;Kim, JinWon;Im, Sua;Shin, Su-Jin;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.30 no.6
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    • pp.522-531
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    • 2020
  • The aim of this study was to establish the optimal medium composition for enhancing L(+)-lactic acid (LLA) production using response surface methodology (RSM). Lactobacillus paracasei SRCM201474 was selected as the LLA producer by productivity analysis from nine candidates isolated from kimchi and identified by 16S rRNA gene sequencing. Plackett-Burman design was used to assess the effect of eleven media components on LLA production, including carbon (glucose, sucrose, molasses), nitrogen (yeast extract, peptone, tryptone, beef extract), and mineral (NaCl, K2HPO4, MgSO4, MnSO4) materials. Glucose, sucrose, molasses, and peptone were subsequently chosen as promising media for further optimization studies, and a hybrid design experiment was used to establish their optimal concentrations as glucose 15.48 g/l, sucrose 16.73 g/l, molasses 39.09 g/l, and peptone 34.91 g/l. The coefficient of determination of the equation derived from RSM regression for LLA production was mathematically reliable at 0.9969. At optimum parameters, 33.38 g/l of maximum LLA increased by 193% when compared with MRS broth as unoptimized medium (17.66 g/l). Our statistical model was confirmed by subsequent validation experiments. Increasing the performance of LLA-producing microorganisms and establishing an effective LLA fermentation process can be of particular benefit for bioplastic technologies and industrial applications.

Statistical Optimization of the Growth Factors for Chaetoceros neogracile Using Fractional Factorial Design and Central Composite Design

  • Jeong, Sung-Eun;Park, Jae-Kweon;Kim, Jeong-Dong;Chang, In-Jeong;Hong, Seong-Joo;Kang, Sung-Ho;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1919-1926
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    • 2008
  • Statistical experimental designs; involving (i) a fractional factorial design (FFD) and (ii) a central composite design (CCD) were applied to optimize the culture medium constituents for production of a unique antifreeze protein by the Antartic micro algae Chaetoceros neogracile. The results of the FFD suggested that NaCl, KCl, $MgCl_2$, and ${Na}_{2}{SiO}_{3}$ were significant variables that highly influenced the growth rate and biomass production. The optimum culture medium for the production of an antifreeze protein from C. neogracile was found to be Kalle's artificial seawater, pH of $7.0{\pm}0.5$, consisting of 28.566 g/l of NaCl, 3.887 g/l of $MgCl_2$, 1.787 g/l of $MgSO_4$, 1.308 g/l of $CaSO_4$, 0.832 g/l of ${K_2}{SO_4}$, 0.124 g/l of $CaCO_3$, 0.103 g/l of KBr, 0.0288 g/l of $SrSO_4$, and 0.0282 g/l of ${H_3}{BO_3}$. The antifreeze activity significantly increased after cells were treated with cold shock (at $-5^{\circ}C$) for 14 h. To the best of our knowledge, this is the first report demonstrating an antifreeze-like protein of C. neogracile.

Statistical Optimization for Monacolin K and Yellow Pigment Production and Citrinin Reduction by Monascus purpureus in Solid-State Fermentation

  • Jirasatid, Sani;Nopharatana, Montira;Kitsubun, Panit;Vichitsoonthonkul, Taweerat;Tongta, Anan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.364-374
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    • 2013
  • Monacolin K and yellow pigment, produced by Monascus sp., have each been proven to be beneficial compounds as antihypercholesterolemic and anti-inflammation agents, respectively. However, citrinin, a human toxic substance, was also synthesized in this fungus. In this research, solidstate fermentation of M. purpureus TISTR 3541 was optimized by statistical methodology to obtain a high production of monacolin K and yellow pigment along with a low level of citrinin. Fractional factorial design was applied in this study to identify the significant factors. Among the 13 variables, five parameters (i.e., glycerol, methionine, sodium nitrate, cultivation time, and temperature) influencing monacolin K, yellow pigment, and citrinin production were identified. A central composite design was further employed to investigate the optimum level of these five factors. The maximum production of monacolin K and yellow pigment of 5,900 mg/kg and 1,700 units/g, respectively, and the minimum citrinin concentration of 0.26 mg/kg were achieved in the medium containing 2% glycerol, 0.14% methionine, and 0.01% sodium nitrate at $25^{\circ}C$ for 16 days of cultivation. The yields of monacolin K and yellow pigment were about 3 and 1.5 times higher than the basal medium, respectively, whereas citrinin was dramatically reduced by 36 times.

Statistical optimization of culture media contained soy proteins and hypocotyl for the growth of Bifidobacterium lactis BL 740 and production of soy isoflavone aglycones (대두 단백질 및 배아를 이용한 Bifidobacterium lactis BL740의 균체성장 및 이소플라본 비배당체 생산를 위한 통계적 배지 최적화)

  • Lee, Choong-Young;Lee, Yoon-Bok;Lee, Keun-Ha;Park, Myeong-Soo;Hwang, Seock-Yeon;Hong, Seung-Bok;Yoo, Yung-Choon;Yu, Byung-Yeon;Kim, Chung-Ho
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.126-131
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    • 2010
  • In order to maximize the growth of Bifidobacterium lactis BL 740 and soy isoflavone agycones production, we investigated the optimization of a culture medium containing soy hypocotyls, which are the byproducts of the soy manufacturing process, and soy proteins. The ingredients of the medium containing soy materials (S-medium) were selected by fractional factorial design (FFD) and central composite design (CCD) within a desirable range. The FFD was applied by six factors: glucose, cellobiose, fructooligosaccharide, soy peptone, soy protein, and soy hypocotyl. Soy protein, soy peptone, and soy hypocotyl were found to be significant factors from the result of FFD for both the growth of B. lactis BL 740 and aglycone production. The CCD was then applied with three variables found from FFD at five levels each and the optimum values were determined for the three variables: soy peptone, soy protein, and soy hypocotyl. In the case of the growth of B. lactics BL740, the proposed optimal media contained 12.73 g/L of soy protein, 29.55 g/L of soy peptone, and 130.67 g/L of soy hypocotyl. To produce isoflavone aglycones, optimized media was composed of 2.06 g/L, soy protein, 1.25 g/L of soy peptone, and 60.02 g/L of soy hypocotyl.