• Title/Summary/Keyword: fermenter culture

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Enhanced Production of Soluble Pyrococcus furiosus α-Amylase in Bacillus subtilis through Chaperone Co-Expression, Heat Treatment and Fermentation Optimization

  • Zhang, Kang;Tan, Ruiting;Yao, Dongbang;Su, Lingqia;Xia, Yongmei;Wu, Jing
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.570-583
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    • 2021
  • Pyrococcus furiosus α-amylase can hydrolyze α-1,4 linkages in starch and related carbohydrates under hyperthermophilic condition (~ 100℃), showing great potential in a wide range of industrial applications, while its relatively low productivity from heterologous hosts has limited the industrial applications. Bacillus subtilis, a gram-positive bacterium, has been widely used in industrial production for its non-pathogenic and powerful secretory characteristics. This study was conducted to increase production of P. furiosus α-amylase in B. subtilis through three strategies. Initial experiments showed that co-expression of P. furiosus molecular chaperone peptidyl-prolyl cis-trans isomerase through genomic integration mode, using a CRISPR/Cas9 system, increased soluble amylase production. Therefore, considering that native P. furiosus α-amylase is produced within a hyperthermophilic environment and is highly thermostable, heat treatment of intact culture at 90℃ for 15 min was performed, thereby greatly increasing soluble amylase production. After optimization of the culture conditions (nitrogen source, carbon source, metal ion, temperature and pH), experiments in a 3-L fermenter yielded a soluble activity of 3,806.7 U/ml, which was 3.3- and 28.2-fold those of a control without heat treatment (1,155.1 U/ml) and an empty expression vector control (135.1 U/ml), respectively. This represents the highest P. furiosus α-amylase production reported to date and should promote innovation in the starch liquefaction process and related industrial productions. Meanwhile, heat treatment, which may promote folding of aggregated P. furiosus α-amylase into a soluble, active form through the transfer of kinetic energy, may be of general benefit when producing proteins from thermophilic archaea.

Optimum culture conditions of cell growth and polysaccharide production by Paecilomyces japonicain batch culture

  • Park, Seok-Jae;Byeon, Hak-Gyu;Han, Dae-Seok;Hong, Eok-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.287-290
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    • 2000
  • To examine effects of agitation and aeration as well as adding of glucose and yeast extract on cell growth and polysaccharide production by Paecilomyces japonica, batch culture was carried out at 5L jar fermenter at $27^{\circ}C$ with the initial pH 7 for 7 days cultivation(innoculum size 2%, working volume 3L). Media compositions(g/L) were 30 glucose, 20 yeast extract, 0.5 $KH_2PO_4$, $0.1\;CuCl_2\;{\cdot}\;2H_2O$. Optimum culture conditions of agitation and aeration in batch culture were 400 rpm and 1.0 vvm, resulting in 23.1 g/L biomass and 2.5 g/L polysaccharide. Additional feeding of glucose and yeast extract with a pulse mode conferred an advantage on cell growth and polysaccharide production with showing the results of 29.2 g/L and 3.3 g/L, respectively.

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Composting of Food Waste by Non-Stirrer Sealed Fermenter and Change of NaCl content in Soil during the Pepper Cultivation (무교반 밀폐형 발효조를 이용한 음식물류폐기물 퇴비화 및 작물재배 중 염분의 함량 변화)

  • Hong, Sung Gil;Chang, Ki Woon;Kwon, Hyuk Young
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.3
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    • pp.82-88
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    • 2005
  • This study was carried out to investigate on the change of NaCl content during the food waste composting and on the safety of food waste compost(FWC) manufactured by the non-stirrer sealed fermenter. Plant culture test with pepper crop was also performed to see the effect of FWC, which was produced by the G co. ltd., on the growth of peper and migration of NaCl in soil. The culture test was performed at the farmland in Chungnam National University. The results were as follows; the NaCl content was gradually accumulated during food waste composting process, probably through water evaporation. Sodium concentration was, however, remarkably decreased at the final stage due to the desalting effect by water which was concentrated on the ceil of the fermentor. The analysis of chemical properties and humidity parameters on the food waste compost revealed that the product is quite a good qualified one. More than 0.5 tons of FWC application on red pepper cultivation caused diminished effect on the yield and the accumulation of salts on soil.

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Optimization of biomass production of Acetobacter pasteurianus SRCM101388 (Acetobacter pasteurianus SRCM101388 바이오매스 생산 최적화)

  • Jun-Tae Kim;Sung-Ho Cho;Do-Youn Jeong;Young-Soo Kim
    • Food Science and Preservation
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    • v.30 no.1
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    • pp.132-145
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    • 2023
  • In this study, culture conditions were optimized to confirm the feasibility of Acetobacter pasteurianus as a starter for fermentation vinegar. Acetobacter pasteurianus strain can be used as a food ingredient. The optimal temperature and pH conditions of the selected Acetobacter pasteurianus SRCM101388 were 28℃ and pH 6.00, respectively. The response surface methodology (RSM) was used to optimize the composition of the medium, and Plackett-Burman design (PBD) was used to obtain the effective selection of culture medium, resulting in that glucose, sucrose, and yeast extract had the highest effect on increasing biomass. The optimal concentration, which was performed by central composite design (CCD), were determined to be 10.73 g/L of glucose, 3.98 g/L of sucrose, and 18.73 g/L of yeast extract, respectively. The optimal concentrations of trace elements for the production of biomass were found to be 1 g/L of ammonium sulfate, 0.5 g/L of magnesium sulfate, 2 g/L of sodium phosphate monobasic, 2 g/L of sodium phosphate dibasic, and the final optimized medium was pH 6.10. When incubated in a 5 L jar fermenter, the SRCM101388 strain showed a faster-dissolved oxygen (DO) reduction at a lower agitation rate (rpm), and it was able to grow even at reduced DO level when aeration was maintained. The amount of final biomass produced was 2.53±0.12×109 CFU/mL (9.40±0.02 log CFU/mL) when incubated for 18 hours at 150 rpm, 0.5 vvm, pH 6.0, and 28℃.

The Anti-complementary Activity of Exo-polymers Produced from Submerged Mycelial Cultures of Higher Fungi with Particular Reference to Cordyceps militaris

  • Song, Chi-Hyun;Jeon, Young-Jae;Yang, Byung-Keun;Ra, Kyung-Soo;Sung, Jae-Mo
    • Journal of Microbiology and Biotechnology
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    • v.8 no.5
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    • pp.536-539
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    • 1998
  • The anti-complementary activity (immuno-stimulating activity) was tested for the exo-polymers (extra-cellular polymer) produced from submerged mycelial cultures of 21 types of higher fungi. Anti-complementary activity of the exo-polymer from Cordyceps militaris showed the highest (69.0%) followed by Pleurotus ostreatus (63.7%) and Trametes suaveolens (61.4%). The mycelial growth rate and biomass doubling time of C. militaris in a 5 I air-lift fermenter were 0.0255 $h^{-1}$ and 27.2 h, respectively. The yield of the exo-polymer produced from the culture broth of C. militaris was 2.95 mg of dry weight/ml of culture broth. Sugar and amino acid compositions of this exo-polymer were analyzed.

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Effect of Different pH Processes on Branched β-1,3-Glucan Production from Submerged Culture of Ganoderma lucidum (영지 (G. lucidum)의 액체배양에 의한 β-1,3-Glucan 생산에 미치는 서로 다른 pH Process의 영향)

  • Lee, Shin-Yaung;Lee, Kyu-Min
    • Journal of Industrial Technology
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    • v.20 no.A
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    • pp.45-50
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    • 2000
  • A submerged cultivation of Ganoderma lucidum was carried out in an air-lift fermenter system, and the effects of different pH processes on extracellular branched ${\beta}$-1,3-glucan(EPS) production and mycelial growth(MDW) were investigated. The controlled pH process improved the production of branched ${\beta}$-1,3-glucan and biomass in comparison to the uncontrolled pH process. However, the maximum production of branched ${\beta}$-1,3-glucan were obtained by the bi-staged pH process. From these results, we confirmed that the bi-staged pH process was the most effective for improving the production of branched ${\beta}$-1,3-glucan from submerged culture of G. lucidum.

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The Optimum Conditions for the Production of Exo-polymer from Submerged Mycelial Culture of Ganoderma lucidum WK-003 and It's Hepatoprotective Effect (Ganoderma lucidum WK-003 균사체 액체배양으로부터 균체외 고분자물질의 생산조건과 간 보호 효과)

  • 송치현;양병근;전용재;나경수;손동환;김혁일;김영환
    • KSBB Journal
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    • v.13 no.4
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    • pp.364-368
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    • 1998
  • The optimum conditions of the production of exo-polymer by Ganoderma lucidum WK-003 and it's hepatoprotective effect was studied. Optimum conditions for the production of exo-polymer (3.18 g/$\ell$) by using shaken flask culture of G. lucidum WK-003 were pH 4.5, 30$^{\circ}C$, 120 rpm for 18 days cultivation. Also exo-polymer productio (7.15 g/$\ell$) was optimized by 5$\ell$ jar fermenter cultivation with condition of pH 4.5, 30$^{\circ}C$, 200 rpm, 1.0 vvm for 6 days cultivation. Glutamic pyruvic transaminase(GPT) activities in serum of intoxicated Sprague-Dawley rat were decreased from 704 IU/L to 330 IU/L by oral administration of the exo-polymer (20mg/kg$.$day)f or 4 consecutive days.

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Favorable Condition for Mycelial Growth of Tricholoma matsutake (송이균 배양을 위한 균사생장 조건)

  • Kim, In-Yeup;Jung, Gwang-Reul;Han, Sang-Kuk;Cha, Joo-Young;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.33 no.1
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    • pp.22-29
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    • 2005
  • The main objectives of this research were to study the cultural and nutritional characteristics of Tricholoma matsutake and to establish its liquid culture system. The optimum growth of T. matsutake was observed in HA and TMM agar media. Similarly highest growth was observed in PDB and TMM liquid media. The optimal temperature for the mycelial growth was $25^{\circ}C$. The most suitable carbon source was dextrin among 12 different carbon sources tested. Yeast extract and peptone were best nitrogen sources among 17 different sources tested. The optimum mineral salts were $Fe_{2}(SO_{4})_{3}{\cdot}H_{2}O$ and KCl among 9 different sources tested. Shaking culture gave higher mycelial growth compared to stationary culture. Similarly, optimum medium amount for shaking culture was 100 ml per 250 ml flask. The highest mycelial growth was obtained when $5{\sim}7$ mycelial discs were inoculated in 100 ml of medium and incubated for $8{\sim}9$ weeks, respectively. The highest proportion of mycelial growth was observed at 40 : 1 ratio of medium to inoculum volume in 8 l air-lift fermenter.

Optimization of Culture Conditions of Chitosanase-producing Bacillus sp. P16 (키토산분해효소 생산을 위한 Bacillus sp. P16 배양조건의 최적화)

  • Jung, Mi-Ra;Jo, Yu-Young;Chil, Youn-Tae;Park, Ro-Dong
    • Applied Biological Chemistry
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    • v.42 no.3
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    • pp.193-198
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    • 1999
  • The optimal culture condition of Bacillus sp. P16 was investigated for production of an extracellular endo-splitting chitosanase. The best carbon and nitrogen sources for the chitosanase production were chitosan and tryptone, respectively. The best condition for the maximum activity was at $37^{\circ}C$ in a medium containing 0.5% powdered chitosan, 1% tryptone, and 1% NaCl(at initial pH 7.0) in a rotary shaker(200 rpm). In a jar fermenter, the culture duration shortened to $6{\sim}12$ hr for maximum activity and the enzyme activity increased about 100% compared with that of flask culture.

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Effect of Galactose and Dextrose on Human Lipocortin I Expression in Recombinant Saccharomyces cerevisiae Carrying Galactose-Regulated Expression System

  • Nam, Soo-Wan;Seo, Dong-Jin;Rhee, Sang-Ki;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.3 no.3
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    • pp.168-173
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    • 1993
  • The expression kinetics of human lipocortin I (LCI), a potential anti-inflammatory agent, was studied in the shake-flask and fermenter cultures of Saccharomyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LCI, and the plasmid stability were investigted under various galactose induction conditions. The expression of LCI was repressed by the presence of a very small amount of dextrose in the culture medium, but it was induced by galactose after dextrose became completely depleted. The optimal ratio of dextrose to galactose for lipocortin I production was found to be 1.0 (10 g/l dextrose and 10 g/l galactose). With optimal D/G ratio of 1.0 and the addition of galactose prior to dextrose depletion, LCI of about 100~130 mg/l was produced. LCI at a concentration of 174 mg/l was porduced in the fed-batch culture, which was nearly a twice as much of that produced in the batch culture. The plasmid stability was very high in all culture cases, and thus was considered to be not an important parameter in the expression of LCI.

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