• Title/Summary/Keyword: submerged cultures

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Hypoglycemic Effects of Exo-biopolymers Produced by Five Different Medicinal Mushrooms in STZ-induced Diabetic Rats

  • Yang, Byung-Keun;Kim, Guk-Nam;Jeong, Yong-Tae;Jeong, Hun;Mehta, Pradeep;Song, Chi-Hyun
    • Mycobiology
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    • v.36 no.1
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    • pp.45-49
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    • 2008
  • Hypoglycemic effects of exo-biopolymers (EBP) produced by submerged mycelial cultures of Coriolus versicolor, Cordyceps sinensis, Paecilomyces japonica, Armillariella mellea, and Fomes fomentarius were investigated in streptozotocin (STZ)-induced diabetic rats. The rats from each experimental group were orally administered with EBPs (100 mg/kg BW) daily for 2 weeks. Though the hypoglycemic effect was achieved in all the cases, however, C. versicolor EBP proved as the most potent one. The administration of the C. versicolor EBP substantially reduced (29.9%) the plasma glucose level as compared to the saline administered group (control). It also reduced the plasma total cholesterol (TC), triglyceride (TG), aspartate aminotransferase (AST) and, alanine aminotransferase (ALT) levels by 9.22, 23.83, 16.93, and 27.31%, respectively. The sugar and amino acid compositions of this EBP were also analyzed in detail.

Kinetic Modeling of Submerged Culture of A. blazei with Mixed Carbon Sources of Glucose and Dextrin

  • Na Jeong-Geol;Kim Hyun-Han;Chang Yong-Keun;Lee Sang-Jong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1331-1337
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    • 2006
  • A mathematical model has been proposed for the batch culture of Agaricus blazei with mixed carbon sources of glucose and dextrin. In the proposed model, the metabolism of A. blazei was divided into three parts: cell growth, exopolysaccharides (EPS) production, and another EPS production pathway activated by dextrin hydrolysis. Each pathway was described mathematically and incorporated into the mechanistic model structure. Batch cultures were carried out with six different carbon source compositions. Although parameters were estimated by using the experimental data from the two extreme cases such as glucose only and dextrin only, the model represented well the profiles of glucose, cell mass, and EPS concentrations for all the six different carbon source mixtures, showing a good interpolation capability. Of note, the lag in EPS production could be quite precisely predicted by assuming that the glucose-to-cell mass ratio was the governing factor for EPS production.

Fungal Biotransformation of Monoterpenes Found in Agro-Industrial Residues from Orange and Pulp Industries into Aroma Compounds: Screening Using Solid Phase Microextraction

  • Junior, Mario Roberto Marostica;Mota, Natasha Onoyama;Baudet, Nathalie;Pastore, Glaucia Maria
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.37-42
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    • 2007
  • The biotransformation of monoterpenic agro-industrial wastes (turpentine oil and essential orange oil) was studied. More than 40 fungal strains were isolated from Brazilian tropical fruits and eucalyptus trees and screened for biotransformation of the waste substrates. Solid phase microextraction was used to monitor the presence of volatile compounds in the headspaces of sporulated surface cultures. The selected strains were submitted to submerged liquid culture. The biotransformation of R-(+)-limonene and ${\alpha},\;{\beta}-$ pinenes from the oils resulted in ${\alpha}-terpineol$ and perillyl alcohol, and verbenol and verbenone, respectively, as the main products. The selected strains were also placed in contact with ${\alpha}-$ and ${\beta}-$ pinenes standards. It was confirmed that verbenol, verbenone, and ${\alpha}-terpineol$ were biotransformation products from the terpenes. A concentration of 90 mg/L of verbenone was achieved by Penicillium sp. 2360 after 3 days of biotransformation.

Calcium Absorption and Growth Characteristics of Agrocybe cylindracea Mycelia in Submerged Culture (액체배양 버들송이 균사체의 칼슘흡수 및 생육특성)

  • Park, Shin;Gong, Ji-Won;Lee, Kyoung-Seok
    • Korean Journal of Food Science and Technology
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    • v.40 no.4
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    • pp.419-423
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    • 2008
  • The present study examined the effects of different calcium (Ca) sources and concentrations on the growth and Ca absorption of Agrocybe cylindracea mycelia grown in submerged cultures. The dry weights of the mycelia were not significantly different (significance level of 5%) according to the type of Ca added, and increased with increasing Ca concentration until 500 mg/L, and then decreased at concentrations of 1000 mg/L or greater. The Ca contents of groups were significantly different according to the various concentrations of the Ca source, in which the Ca content of the control group cultured without added Ca was 198.3 mg/kg, and in the treatment groups, Ca content increased to a minimum of 273.7 mg/kg (1.4 times) and a maximum of 67246.0 mg/kg (339.1 times) the Ca contents of the groups generally increased with increasing Ca concentration. According to the number of culture days, growth rates were highest during days 8 through 12, and remained relatively high until day 16. In addition, Ca contents per unit dry weight were higher in young mycelia with a shorter culture period than in mature mycelia with a longer culture period. According to pH, the most active growth and highest Ca content occurred in MCM liquid medium at pH 7.0. In conclusion, in order to produce Agrocybe cylindracea mycelia with high Ca content, it is considered most efficient to culture them in MCM liquid medium without a pH adjustment and containing 1,000 mg/L of Ca-lactate, which is commonly used as a Ca additive in food, as well as to use mycelia between 12-16 days of culturing.

Kinetic Studies on Submerged Acetic Acid Fermentation of Acetobacter aceti (Acetobacter aceti균의 심부배양에 의한 초산발효의 동력학적 연구)

  • KIM Sang-Moo;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.4
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    • pp.301-306
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    • 1992
  • In order to determine the optimal conditions for the process of acetic acid fermentation, the kinetics of Acetobacter aceti fermentation in submerged batch cultures were studied at different agitation speeds and air flow rates. The maximum cell concentration was noted after about 48 hr fermentation and the time course of Acetobactey aceti fermentation showed a distinct feature of growth-associated product formation. At agitation speeds 700, 500, and 300 rpm fixed on air flow rate 1 v/v/M, specific grow rates were $3.97\times10^{-2},\;3.82\times10^{-2},\;and\;2.04\times10^{-2\} \;hr^{-1}$, saturation constants were 61.4, 64.6, and 69.4mg/ml. and volumetric oxygen transfer coefficients were 0.9337, 0.4468, and 0.1701 $min^{-1},$ respectively. At air flow rates 1.25, 1.00, and 0.75 v/v/M fixed on agitation speed 500 rpm, specific growth rates were $3.90\times10^{-2},\;3.82\times10^{-2},\;and\;2.37\times10^{-2}\;hr^{-1}$, saturation constants were 63.4, 64.6, and 64.9 mg/ml, and volumetric oxygen transfer coefficient were 0.4923, 0.4468, and 0.3509 $min^{-1},$ respectively.

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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.

Characterization of Homocysteine ${\gamma}$-Lyase from Submerged and Solid Cultures of Aspergillus fumigatus ASH (JX006238)

  • El-Sayed, Ashraf S.;Khalaf, Salwa A.;Aziz, Hani A.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.499-510
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    • 2013
  • Among 25 isolates, Aspergillus fumigatus ASH (JX006238) was identified as a potent producer of homocysteine ${\gamma}$-lyase. The nutritional requirements to maximize the enzyme yield were optimized under submerged (SF) and solid-state fermentation (SSF) conditions, resulting in a 5.2- and 2.3-fold increase, respectively, after the last purification step. The enzyme exhibited a single homogenous band of 50 kDa on SDS-PAGE, along with an optimum pH of 7.8 and pH stability range of 6.5 to 7.8. It also showed a pI of 5.0, as detected by pH precipitation with no glycosyl residues. The highest enzyme activity was obtained at $37-40^{\circ}C$, with a $T_m$ value of $70.1^{\circ}C$. The enzyme showed clear catalytic and thermal stability below $40^{\circ}C$, with $T_{1/2}$ values of 18.1, 9.9, 5.9, 3.3, and 1.9 h at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. Additionally, the enzyme $K_r$ values were 0.002, 0.054, 0.097, 0.184, and 0.341 $S^{-1}$ at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. The enzyme displayed a strong affinity to homocysteine, followed by methionine and cysteine when compared with non-S amino acids, confirming its potency against homocysteinuria-related diseases, and as an anti-cardiovascular agent and a specific biosensor for homocysteinuria. The enzyme showed its maximum affinity for homocysteine ($K_m$ 2.46 mM, $K_{cat}\;1.39{\times}10^{-3}\;s^{-1}$), methionine ($K_m$ 4.1 mM, $K_{cat}\;0.97{\times}10^{-3}\;s^{-1}$), and cysteine ($K_m$ 4.9 m M, $K_{cat}\;0.77{\times}10^{-3}\;s^{-1}$). The enzyme was also strongly inhibited by hydroxylamine and DDT, confirming its pyridoxal 5'-phosphate (PLP) identity, yet not inhibited by EDTA. In vivo, using Swiss Albino mice, the enzyme showed no detectable negative effects on platelet aggregation, the RBC number, aspartate aminotransferase, alanine aminotransferase, or creatinine titer when compared with negative controls.

Mechanism of Free Phosphate Production by Penicillium sp. GL-101, Phosphate Solubilizing Fungus, in the Submerged Culture (인산가용화균 Penicillium sp. GL-101의 유리인산 생성기작에 관한 연구)

  • Kang, Sun-Chul;Yang, Mi-Ok;Tae, Un-Hee
    • Korean Journal of Environmental Agriculture
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    • v.20 no.1
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    • pp.1-7
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    • 2001
  • We investigated the capability of the phosphate-solubilizing fungus, Penicillium sp. GL-101, to solubilize in vitro some insoluble rock phosphate via possible mechanisms: acidification of the medium, production of chelating metabolites, redox activity, and so on. GL-101 was able to solubilize rock phosphate (mostly calcium phosphate) in a liquid potato dextrose broth(PDB) medium, as determined by spectrophotometric analyses. Acidification was the major mechanism of solubilization since the pH of cultures fell below 4.0 and in cultures containing 1.0%(w/v) loess the pH dropped from 7.0 to 3.2. More than 10 mg/mL concentrations of citric acids were detected by high-performance liquid chromatography(HPLC) in the culture supernatants. Also this fungus showed the phosphatase activity (over 1.3 unit) to contribute partially releasing phosphate from rock phosphate, when supplemented with 1.0% loess in culture broth. The chelating activity of GL-101 in culture supernatants was not present because 2-ketogluconic acid, a chelating agent for the phosphate, was produced only a basal level. Therefore, the solubilization mechanism of rock phosphate by Penicillium sp. GL-101 involves both acidification due to citric acid production and phosphatase activity.

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Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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Enhanced Production of Human Serum Albumin by Fed-Batch Culture of Hansenula polymorpha with High-Purity Oxygen

  • Youn, Jong-Kyu;Shang, Longan;Kim, Moon-Il;Jeong, Chang-Moon;Chang, Ho-Nam;Hahm, Moon-Sun;Rhee, Sang-Ki;Kang, Hyun-Ah
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1534-1538
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    • 2010
  • Fed-batch cultures of Hansenula polymorpha were studied to develop an efficient biosystem to produce recombinant human serum albumin (HSA). To comply with this purpose, we used a high-purity oxygen-supplying strategy to increase the viable cell density in a bioreactor and enhance the production of target protein. A mutant strain, H. polymorpha GOT7, was utilized in this study as a host strain in both 5-l and 30-l scale fermentors. To supply high-purity oxygen into a bioreactor, nearly 100% high-purity oxygen from a commercial bomb or higher than 93% oxygen available in situ from a pressure swing adsorption (PSA) oxygen generator was employed. Under the optimal fermentation of H. polymorpha with highpurity oxygen, the final cell densities and produced HSA concentrations were 24.6 g/l and 5.1 g/l in the 5-l fermentor, and 24.8 g/l and 4.5 g/l in the 30-l fermentor, respectively. These were about 2-10 times higher than those obtained in air-based fed-batch fermentations. The discrepancies between the 5-l and 30-l fermentors with air supply were presumably due to the higher contribution of surface aeration over submerged aeration in the 5-l fermentor. This study, therefore, proved the positive effect of high-purity oxygen in enhancing viable cell density as well as target recombinant protein production in microbial fermentations.