• Title/Summary/Keyword: Glucose Kinetics

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Altering UDP-Glucose Donor Substrate Specificity of Bacillus licheniformis Glycosyltransferase towards TDP-Glucose

  • Cho, Kye Woon;Kim, Tae-Su;Le, Tuoi Thi;Nguyen, Hue Thi;Oh, So Yeong;Pandey, Ramesh Prasad;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.268-273
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    • 2019
  • The specificity of a Bacillus licheniformis uridine diphosphate (UDP) glycosyltransferase, YjiC, was increased towards thymidine diphosphate (TDP)-sugar by site-directed mutagenesis. The Arg-282 of YjiC was identified and investigated by substituting with Trp. Conversion rate and kinetic parameters were compared between YjiC and its variants with several acceptor substrates such as 7-hydroxyflavone (7-HF), 4',7-dihydroxyisoflavone, 7,8-dihydroxyflavone and curcumin. Molecular docking of TDP-glucose and 7-HF with YjiC model showed pi-alkyl interaction with Arg-282 and His-14, and pi-pi interaction with $His^{14}$ and thymine ring. YjiC (H14A) variant lost its glucosylation activity with TDP-glucose validating significance of His-14 in binding of TDP-sugars.

Batch and Continuous Culture Kinetics for Production of Carotenoids by ${\beta}$-Ionone-Resistant Mutant of Xanthophyllomyces dendrorhous

  • Park, Ki-Moon;Song, Min-Woo;Kang, Seog-Jin;Lee, Jae-Heung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1221-1225
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    • 2007
  • A ${\beta}$-ionone-resistant mutant strain isolated from the red yeast Xanthophyllomyces dendrorhous KCTC 7704 was used for batch and continuous fermentation kinetic studies with glucose media in a 2.5-1 jar fermentor at $22^{\circ}C$ and pH 4.5. The kinetic pattern of growth and carotenoid concentration in the batch fermentations exhibited a so-called mixed-growth-associated product formation, possibly due to the fact that the content of intracellular carotenoids depends on the degree of physical maturation toward adulthood. To determine the maximum specific growth rate constant (${\mu}_m$) and Monod constant ($K_s$) for the mutant, glucose-limited continuous culture studies were performed at different dilution rates within a range of $0.02-0.10\;h^{-1}$. A reciprocal plot of the steady-state data (viz., reciprocal of glucose concentration versus residence time) obtained from continuous culture experiments was used to estimate a ${\mu}_m$ of $0.15\;h^{-1}$ and $k_s$ of 1.19 g/l. The carotenoid content related to the residence time appeared to assume a typical form of saturation kinetics. The maximum carotenoid content ($X_m$) for the mutant was estimated to be $1.04\;{\mu}g/mg$ dry cell weight, and the Lee constant ($k_m$), which was tentatively defined in this work, was found to be 3.0 h.

Fermentation Kinetics for Production of Carotenoids by ${\beta}$-ionone Resistant Mutant of Xanthophyllomyces dendrorhous (Xanthophyllomyces dendrorhous 변이군주에 의한 Carotenoids 생산 발효의 특성 연구)

  • Park, Ki-Moon;Kim, Young-Jun;Song, Min-Woo;Kang, Seog-Jin;Lee, Jae-Heung
    • KSBB Journal
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    • v.21 no.4
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    • pp.286-291
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    • 2006
  • Various ${\beta}$-ionone resistant mutants were isolated from the wild-type red yeast Xanthophyllomyces dendrorhous KCTC 7704. Although the growth of X. dendrorhous KCTC 7704 was strongly inhibited at 0.025 mM ${\beta}$-ionone, one of the ${\beta}$-ionone resistant mutants isolated at 0.1 mM ${\beta}$-ionone by NTG mutagenesis showed rather 70% of relative survival at 0.15 mM ${\beta}$-ionone. Fermentation kinetics study with the mutant was carried out at $20^{\circ}C$ for 4 days in 300-mL baffled flasks. The mutant yielded up to 2.3-fold higher carotenoids content(viz. $1.2{\mu}g$ of total carotenoids per mg of dry cells) compared with the wild-type strain. The production of metabolites such as organic acids could be neglected. Studies on the kinetics with various carbon substrates revealed both an increase in final dry cell mass and a higher total carotenoids content in cell mass with glucose when compared to fructose or sucrose. As a further part of study, the effect of pH on the fermentation kinetics was investigated in glucose-limited chemostat at a dilution rate of $0.04h^{-1}$. When compared to steady-state kinetic parameters obtained at pH 4.0, a significant reduction in cell concentration at pH 3.0 and a lower carotenoids content at pH 5.2 were evident.

Nutrient Balance and Glucose Metabolism of Female Growing, Late Pregnant and Lactating Etawah Crossbred Goats

  • Astuti, D.A.;Sastradipradja, D.;Sutardi, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.8
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    • pp.1068-1075
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    • 2000
  • A study involving nutrient balances and radioisotope labeling techniques was undertaken to study energy and protein metabolism, and glucose kinetics of female crossbred Etawah goats, using 12 weaned (BW $14.0{\pm}2.0kg$), 12 late pregnant (BW $27.8{\pm}1.8kg$) and 12 first lactation does (BW $25.0{\pm}5.0kg$). Each class of animal was randomly allotted into 3 dietary treatment groups R1, R2 and R3, that received 100%, 85%, and 70% of ad libitum feed. The rations offered were pellets containing 21.8% CP and 19.3 MJ GE/kg, except for the lactating does who received pellets (17.2% CP and 18.9 MJ GE/kg) and fresh Penisetum purpureum grass. Energy and nitrogen balance studies were conducted during a two-week trial. Daily heat production (HP, estimated by the carbon dioxide entry rate technique), glucose pool and flux were measured. Equations were found for metabolizable energy (ME) and protein intake (IP) requirements for growing goats: ME (MJ/d)=1.87+0.55 RE-0.001 ADG+0.044 RP $(R^2=0.89)$ and IP (g/d)=48.47+2.99 RE+0.029 ADG+0.79 RP $(R^2=0.90)$; for pregnant does: ME (MJ/d)=5.92+0.96 RE-0.002 ADG+0.003 RP $(R^2=0.99)$ and IP (g/d)=58.34+5.41 RE+0.625 ADG-0.30 RP $(R^2=0.98)$; and for lactating does: ME (MJ/d)=4.23+0.713 RE+0.003 ADG+0.006 RP+0.002 MY $(R^2=0.86)$; IP (g/d)=84.05-5.36 RE+0.055 ADG-0.16 RP+0.068 MY $(R^2=0.45)$, where RE is retained energy (MJ/d), ADG is average daily gain in weight (g/d), RP is retained protein (g/d) and MY is milk yield (ml/d). ME and IP requirements for maintenance for growing goats were 0.46 MJ/d.kg $BW^{0.75}$ and 7.43 g/d.kg $BW^{0.75}$, respectively. Values for the pregnant and lactating does were in the same order, 0.55 MJ/d.kg $BW^{0.75}$ and 11.7 g/d.kg $BW^{0.75}$, and 0.50 MJ/d.kg $BW^{0.75}$ and 10.8 g/d.kg $BW^{0.75}$, respectively. Milk protein ranged from 3.06 to 3.5% and milk fat averaged 5.2%. Glucose metabolism in Etawah crossbred female goat is active, but glucose flux is low compared to temperate ruminant breeds which may implicate its role to support production.

Batch Kinetics of Exo-polysaccharide Production by Submerged Cultivation of Ganoderma lucidum (영지의 액체배양에 의한 세포외 다당 생산의 동력학적 특성)

  • Lee, Shin-Young;Lee, Hak-Su;Park, Heung-Cho
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.304-311
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    • 1999
  • Batch kinetics during the exo-polysaccharide (EPS) fermentation of Ganoderma lucidum was investigated as a function of different substrates (glucose and starch), substrate concentration $(1{\sim}7%,\;w/v)$ and subculture (3 times). Logistic model for mycelial growth fitted the experimental data better than Monod and two thirds power model. The Luedeking-Pirt equation was adequate to fit the kinetic data of product formation and substrate consumption. The EPS production was strongly non-growth associated, although it was mixed type. The product formation and sustrate consumption by growth associated mechanism decreased as the concentration of glucose increased, while those of the non-growth associated mechanism increased. However, starch medium increased the growth associated and non-growth associated substrate consumption indicating higher availability of substrate. Also, batch culture in starch medium showed the higher specific growth rate and stability during subculture than those in glucose medium. In conclusion, the enhanced EPS production and stability in the subculture was found to be remarkably improved by use of starch as sole carbon source in medium. The maximum mycelium dry weight and EPS production of 9.463 and 10.410 g/l, respectively, were obtained after shake culture of 7 days at $30^{\circ}C$ from the media containing 7% starch.

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A Kinetic Modeling for the Dynamics of Hybridoma Cells in Suspension Culture (현탁배양 하이브리도마 세포의 속도론적 모델링)

  • 정연호;박현규최정우
    • KSBB Journal
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    • v.11 no.3
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    • pp.276-287
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    • 1996
  • Batch suspension cultures of hybridoma cell were performed with various initial glutamine concentrations to investigate the effects of glutamine on cell growth and death, monoclonal antibody production, glucose and glutamine consumption, and the production of lactate and ammonium ion. An mathematical kinetic model was formulated to describe the kinetics of cell growth, the consumption of nutrients (glucose and glutamine), and the production of monoclonal antibody and waste metabolites (lactate and ammonium ion) based on experimental data. An equation for the specific growth rate was developed such that superimposed Monod equation in glucose and glutamine, with non-competitive type inhibition relations in ammonium ion and lactate. The inhibition constant for lactate was inversely proportional to the lactate concentration. The specific death rate was considered to be a function of glucose, glutamine, ammonium ion and lactate concentration.

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Mathematical Model for the Production of High-purity Fructo-oligosaccharides by the Mixed-enzyme System of Fructosyltransferase and Glucose Oxidase (Fructosyltransferase와 Glucose oxidase 혼합효소계를 이용한 고순도 Fructo-oligosaccharides 생산에서 반응 메카너즘에 대한 수학적 모델)

  • 윤종원;최윤찬이민규송승구
    • KSBB Journal
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    • v.9 no.1
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    • pp.40-47
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    • 1994
  • A simplified mathematical model for the production of high-purity fructo-oligosaccharides by the mixed-enzyme system of fructosyl transferees and glucose oxidase was proposed and compared with the experimental results. The kinetic parameters including $K_m,\;V_{max}\;and\;K_{iG}$ were estimated at $40^{\circ}C$, in which $K_m$, values decreased and $K_{iG}$ and $V_{max}$ values increased compared with those of fructosyl transferees alone. The kinetics of the mixed-enzyme system was successfully described in the form of Michaelis-Menten equations. At the reasonable sucrose concentrations tested, the simulated sugar profiles were of good agreement with the experimental ones.

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Effect of Sugar Starvation on the Sugar Transport System in Suspension Cultures of Streptanthus trotus (Streptanthus tortus 배양 세포에서 당류고갈이 당류 수송계에 미치는 영향)

  • 조봉희
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.47-50
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    • 2000
  • In suspension cultures of Streptanthus. the uptake rate of sugar was increased during the ceil starvation of sugar in the medium. The maximal uptake rate obtained with 3 days of cell starvation. Sugar transport system induced by the sugar starvation was completely inhibited by 10 $\mu$M cycloheximide. Plant cells are known to possess only one sugar transport system, but the uptake rate of glucose obtained a saturated kinetic while the one of sucrose had two different kinetics after the sugar starvation. Induced sugar transport systems had different kinetics compared to plant cell. These results showed that higher plants have adaptable ability to induce new sugar transport systems when the environment changed unsuitable.

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Cybernetic Modeling of Simultaneous Saccharification and Fermentation for Ethanol Production from Steam-Exploded Wood with Brettanomyces custersii

  • Shin Dong-Gyun;Yoo Ah-Rim;Kim Seung-Wook;Yang Dae-Ryook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1355-1361
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    • 2006
  • The simultaneous saccharification and fermentation (SSF) process consists of concurrent enzymatic saccharification and fermentation. In the present cybernetic model, the saccharification process, which is based on the modified Michaelis-Menten kinetics and enzyme inhibition kinetics, was combined with the fermentation process, which is based on the Monod equation. The cybernetic modeling approach postulates that cells adapt to utilize the limited resources available to them in an optimal way. The cybernetic modeling was suitable for describing sequential growth on multiple substrates by Brettanomyces custersii, which is a glucose- and cellobiose-fermenting yeast. The proposed model was able to elucidate the SSF process in a systematic manner, and the performance was verified by previously published data.

Modeling the effect of mass transfer on the kinetics of fructo-oligosaccharide production by immobilized cells (고정화 세포에 의한 Fructo-oligosaccharides 생산 반응메카니즘에서 물질전달 영향에 대한 수학적 모델)

  • 윤종원;전영중이민규송승구
    • KSBB Journal
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    • v.8 no.3
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    • pp.266-271
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    • 1993
  • The effective diffusion coefficients of glucose, sucrose and fructo-oligosaccharides in Ca-alginate gel beads at high concentration of sucrose solutions were investigated at $50^{\circ}C$. A mathematical model for the kinetics of fructo-oligosaccharide production using immobilized cells was proposed and compared with experimental results varying the bead size, the substrate concentration and the bead ratio. Very low values of diffusion coefficients ranging $1.2-7.6\times10^{-7}\textrm{cm}^2$/sec were obtained, and the predicted results were in good agreement with experimental ones in all cases tested.

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