• Title/Summary/Keyword: fermentation optimization

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Optimization of γ-Aminobutyric Acid (GABA) Production Using Immobilized Lactobacillus plantarum K154 in Submerged Culture of Ceriporia lacerata (Ceriporia lacerata 배양액과 고정화 Lactobacillus plantarum K154를 이용한 감마아미노뷰티르산 생산 최적화)

  • Lee, Eun-Ji;Lee, Sam-Pin
    • Korean Journal of Food Science and Technology
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    • v.47 no.4
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    • pp.438-445
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    • 2015
  • The production of GABA was optimized by co-cultivation of immobilized Lactobacillus plantarum K154 (ILK) with Ceriporia lacerata cultures. The mycelial culture of C. lacerata was performed in a defined medium containing 3% glucose, 3% soybean flour, and 0.15% $MgSO_4$ in a submerged condition for 7 days at $25^{\circ}C$, resulting in the production of 29.7 g/L mycelia, 3.1 g/L exopolysaccharides, 2% (w/w) ${\beta}$-glucan, 68.96 unit/mL protease, and 10.37 unit/mL ${\alpha}$-amylase. ILK in C. lacerata culture showed viable cell counts of $3.13{\time}10^9CFU/mL$ for immobilized cells and $1.48{\time}10^8CFU/mL$ for free cells after 1 day. GABA production in the free and immobilized cells was 9.96 mg/mL and 6.30 mg/mL, respectively, after 7 days. A recycling test of ILK in the co-fermentation was consequently performed five times at $30^{\circ}C$ for 15 days, resulting in the highest production of GABA. GABA could also be efficiently overproduced by co-cultivation with the produced polysaccharides, ${\beta}$-glucan, peptides, and probiotics.

Optimization of Ammonia Percolation Process for Ethanol Production from Miscanthus Sinensis (억새를 이용한 바이오 에탄올 생산을 위한 암모니아 침출 공정 최적화)

  • Kim, Kyoung-Seob;Kim, Jun Seok
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.704-711
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    • 2010
  • Lignocellulose ($2^{nd}$ generation) is difficult to hydrolyze due to the presence of lignin and the technology developed for cellulose fermentation to ethanol is not yet economically viable. However, recent advances in the extremely new field of biotechnology for the ethanol production are making it possible to use of agriculture residuals and nonedible crops biomass, e.q., rice straw and miscanthus sinensis, because of their several superior aspects as agriculture residual and nonedible crops biomass; low lignin, high contents of carbohydrates. In this article, as the basic study of AP(Ammonia Percolation), the properties and the optium conditions of process were established, and then the overall efficiency of AP was investigated. The important independent variables for AP process were selected as ammonia concentration, reaction temperature, and reaction time. The percolation condition for maximizing the content of cellulose, the enzymatic digestibility, and the lignin removal was optimized using RSM(Response Surface Methodology). The determined optimum condition is ammonia concentration; 11.27%, reaction temperature; $157.75^{\circ}C$, and reaction time; 10.01 min. The satisfying results were obtained under this optimized condition, that is, the results are as follows: cellulose content(relative); 39.98%, lignin content(relative); 8.01%, and enzymatic digestibility; 85.89%.

Optimal Production of Xylooligosaccharide by Using Recombinant Endoxylanase from Bacillus subtilis (Bacillus subtilis 유래 재조합 endoxylanase를 이용한 xylooligosaccharide의 최적 생산)

  • Kim, Yeon-Hee;Heo, Sun-Yeon;Kim, Mi-Jin;Lee, Jae-Hyung;Kim, Young-Man;Nam, Soo-Wan
    • Journal of Life Science
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    • v.18 no.1
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    • pp.52-57
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    • 2008
  • Xylan is a major hemicellulose component of the cell walls of monocots and hardwood, representing up to 30% of the dry weight of these plants. To efficiently hydrolyze xylan, the endoxylanase gene from Bacillus sp. was expressed in B. subtilis DB431 by introducing the plasmid pJHKJ4. The total activity of the recombinant endoxylanase reached about 857 unit/ml by batch fermentation of B. subtilis DB431/pJHKJ4 in LB maltose medium. The majority (>92%) of endoxylanase was efficiently secreted into the culture medium. The recombinant endoxylanase hydrolyzed more the birchwood xylan efficiently than the other xylans. When 4 % concentration of xylan was used, the highest production of xylooligosaccharide was observed, and xylobiose and xylotriose were the major products. Optimal amount of enzyme and reaction time for producing xylooligosaccharide were found to be 10 unit and 1 hr, respectively. In addition, the temperature of $40^{\circ}C{\sim}50^{\circ}C$ gave the highest production of xylooligosaccharide. Consequently, the optimized conditions for the production of xylooligosaccharide through the hydrolysis of xylan were determined as follows: 10 unit endoxylanase, $50^{\circ}C$, 4% birchwood xylan, 1 hr reaction.

Evaluation of Optimal Condition for Recombinant Bacterial Ghost Vaccine Production with Four Different Antigens of Streptococcus iniae-enolase, GAPDH, sagA, piaA (연쇄구균증 항원-enolase, GAPDH, sagA, piaA에 대한 재조합 고스트 박테리아 백신의 생산 최적화)

  • Ra, Chae-Hun;Kim, Yeong-Jin;Son, Chang-Woo;Jung, Dae-Young;Kim, Sung-Koo
    • Journal of Life Science
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    • v.19 no.7
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    • pp.845-851
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    • 2009
  • A vector harboring double cassettes; a heterologous gene expression cassette of pHCE-InaN-antigen and a ghost formation cassette of pAPR-cI-E lysis 37 SDM was constructed and introduced to E. coli DH5a. For the production of a bacterial ghost vaccine, bacterial ghosts from E. coli / Streptococcus iniae with four different types of antigens - enolase, GAPDH, sagA and piaA - were produced by the optimization of fermentation parameters such as a glucose concentration of 1 g/l, agitation of 300 rpm and aeration of 1 vvm. Efficiency of ghost bacteria formation was evaluated with cultures of OD$_{600}$=1.0, 2.0 and 3.0. The efficiency of the ghost bacteria formation was 99.54, 99.67, 99.99 and 99.99% with inductions at OD$_{600}$=3.0, 1.0, 2.0 and 1.0 for E. coli/S. iniae antigens enolase, piaA, GAPDH and sagA, respectively. Ghost bacteria as a vaccine was harvested by centrifugation. The antigen protein expressions were analyzed by SDS-PAGE and western blot analysis, and the molecular weights of the enolase, piaA, GAPDH and sagA were 78, 26, 67 and 26 kDa, respectively. The molecular weights of the expressed antigens were consistent with theoretical sizes obtained from the amino acid sequences.

Screening of Lactic Acid Bacteria for Strong Folate Synthesis and Optimization of Fermentation (고엽산 생산능의 유산균 탐색 및 발효 조건 최적화)

  • Du, Kyung Min;Park, Se Jin;Park, Myung Soo;Ji, Geun Eog
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.328-333
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    • 2014
  • Folate is a water-soluble vitamin B that is required for the synthesis of amino acids and nucleic acids. It plays an important role in cell division and cell growth in several living organisms. The purpose of this study was to screen strong folate-synthesizing bacteria and to optimize their culture conditions for folate production. Folate production was quantified by microbiological assays by using folate-dependent strain Lactobacillus rhamnosus KCTC 3237. Folate derivatives were identified by LC-MS/MS. Of the 65 strains of bifidobacteria and lactobacilli tested, L. plantarum Fol 708 demonstrated the greatest ability to produce folate. Its optimal pH for folate production was 5.5 in a pH-controlled, lab-scale fermenter. Coculturing L. plantarum Fol 708 with L. brevis GABA 100 in a milk medium enhanced the level of folate produced in comparison to culturing L. plantarum Fol 708 alone.

Optimum Conditions for The Taste of Kanjang Fermented with Barley Bran (보리등겨로 제조한 간장의 최적 발효 조건)

  • Kwon, O-Jun;Son, Dong-Hwa;Choi, Ung-Kyu;Lee, Suk-Il;Im, Moo-Hyeg;Cho, Young-Je;Yang, Sung-Ho;Kim, Sung-Hong;Chung, Yung-Gun
    • Korean Journal of Food Science and Technology
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    • v.33 no.5
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    • pp.596-602
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    • 2001
  • This study was conducted to find out optimum conditions of kanjang fermented with barley bran. Fermentation conditions for optimization of taste of kanjang made with barley bran was investigated with response surface methodology. Optimal conditions of salty taste was as follows, the content of meju: 15%, the content of salt: 7%, initial temperature: $24^{\circ}C$, middle temperature: $29^{\circ}C$ and end temperature: $14^{\circ}C$. Optimal conditions of palatable taste was as follows, the content of meju: 24%, the content of salt: 13%, initial temperature: $26^{\circ}C$, middle temperature: $25^{\circ}C$ and end temperature: $20^{\circ}C$. Optimal conditions of bitter taste was as follows, the content of meju: 28%, the content of salt: 18%, initial temperature: $35^{\circ}C$, middle temperature: $10^{\circ}C$ and end temperature: $38^{\circ}C$. Optimal conditions of overall acceptability was as follows, the content of meju: 15%, the content of salt: 19%, initial temperature: $30^{\circ}C$, middle temperature: $38^{\circ}C$ and end temperature: $23^{\circ}C$.

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Optimization of an Industrial Medium and Culture Conditions for Probiotic Weissella cibaria JW15 Biomass Using the Plackett-Burman Design and Response Surface Methodology

  • Yu, Hyung-Seok;Lee, Na-Kyoung;Kim, Won-Ju;Lee, Do-Un;Kim, Jong-Ha;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.630-637
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    • 2022
  • The objective of this study was to optimize industrial-grade media for improving the biomass production of Weissella cibaria JW15 (JW15) using a statistical approach. Eleven variables comprising three carbon sources (glucose, fructose, and sucrose), three nitrogen sources (protease peptone, yeast extract, and soy peptone), and five mineral sources (K2HPO4, potassium citrate, ⳑ-cysteine phosphate, MgSO4, and MnSO4) were screened by using the Plackett-Burman design. Consequently, glucose, sucrose, and soy peptone were used as significant variables in response surface methodology (RSM). The composition of the optimal medium (OM) was 22.35 g/l glucose, 15.57 g/l sucrose, and 10.05 g/l soy peptone, 2.0 g/l K2HPO4, 5.0 g/l sodium acetate, 0.1 g/l MgSO4·7H2O, 0.05 g/l MnSO4·H2O, and 1.0 g/l Tween 80. The OM significantly improved the biomass production of JW15 over an established commercial medium (MRS). After fermenting OM, the dry cell weight of JW15 was 4.89 g/l, which was comparable to the predicted value (4.77 g/l), and 1.67 times higher than that of the MRS medium (3.02 g/l). Correspondingly, JW15 showed a rapid and increased production of lactic and acetic acid in the OM. To perform a scale-up validation, batch fermentation was executed in a 5-l bioreactor at 37℃ with or without a pH control at 6.0 ± 0.1. The biomass production of JW15 significantly improved (1.98 times higher) under the pH control, and the cost of OM was reduced by two-thirds compared to that in the MRS medium. In conclusion, OM may be utilized for mass producing JW15 for industrial use.

Establishment of Miniaturized Cultivation Method for Large and Rapid Screening of High-yielding Monascus Mutants, and Enhanced Production of Monacolin-K through Statistical Optimization of Production Medium (Monascus 균사체의 소규모 배양을 통한 고생산성 균주의 대규모 선별방법 확립과 통계적 생산배지 최적화를 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.305-312
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    • 2007
  • It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.

Optimization of Culture Conditions for D-Tagatose Production from D-Galactose by Enterobacter agglomerans. (Entrobacter agglomerans에 의한 D-Galactose로부터 D-Tagatose 생산조건의 최적화)

  • 오덕근;노회진;김상용;노봉수
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.250-256
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    • 1998
  • D-Tagatose production from D-galactose was investigated using 35 type strains of American Culture Type Collection (ATCC) and Korean Collection for Type Cultures (KCTC) which have potential to produce D-tagatose. Enterobacter agglomerans ATCC 27987 was selected as a D-tagatose producing strain due to its short fermentation time and high production of D-tagatose. Optimization of the culture conditions for D-tagatose production by E. agglomerans ATCC 27987 was performed. Among various carbon sources, D-galactose was the most effective carbon source for D-tagatose production. As the D-galactose concentration was increased, cell growth and D-tagatose production increased. Effect of nitrogen sources on D-tagatose production was studied. Of inorganic nitrogen sources, ammonium sulfate was effective one for D-tagatose production and yeast extract was the most suitable organic nitrogen nutrient. The concentrations of inorganic compounds such as KH$_2$PO$_4$, K$_2$HPO$_4$, and MgSO$_4$$.$7H$_2$O were also optimized for D-tagatose production. The optimal medium was determined to contain D-galactose of 20 g/l, yeast extract of 5.0 g/l, (NH$_4$)$_2$SO$_4$ of 2.0 g/l, KH$_2$PO$_4$ of 5.0 g/l, K$_2$HPO of 5.0 g/l, and MgSO$_4$$.$7H$_2$O of 5 mg/l. The optimal environmental conditions in a 250-$m\ell$ flask were found to be pH of 6.0, temperature of 30$^{\circ}C$, and agitation speed of 150 rpm. D-tagatose of 0.41 g/l could be obtained in 24 h from 20 g/l D-galactose at the optimal culture condition without induction and cell concentration.

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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.