scholarly journals Flux Analysis of the Metabolism ofClostridium cellulolyticum Grown in Cellulose-Fed Continuous Culture on a Chemically Defined Medium under Ammonium-Limited Conditions

2001 ◽  
Vol 67 (9) ◽  
pp. 3846-3851 ◽  
Author(s):  
Mickaël Desvaux ◽  
Henri Petitdemange
Keyword(s):  
Cellulose Degradation ◽  
Lactate Production ◽  
Nitrogen Sources ◽  
Maximum Growth ◽  
Defined Medium ◽  
Flux Analysis ◽  
Specific Rate ◽  
Cellulolytic Bacteria ◽  
Limiting Nutrient ◽  
Mesophilic Bacterium

ABSTRACT An investigation of cellulose degradation by the nonruminal, cellulolytic, mesophilic bacterium Clostridium cellulolyticum was performed in cellulose-fed chemostat cultures with ammonium as the growth-limiting nutrient. At any dilution rate (D), acetate was always the main product of the catabolism, with a yield of product from substrate ranging between 37.7 and 51.5 g per mol of hexose equivalent fermented and an acetate/ethanol ratio always higher than 1. AsD rose, the acetyl coenzyme A was rerouted in favor of ethanol pathways, and ethanol production could represent up to 17.7% of the carbon consumed. Lactate was significantly produced, but with increasing D, the specific lactate production rate declined, as did the specific rate of production of extracellular pyruvate. The proportion of the original carbon directed towards phosphoglucomutase remained constant, and the carbon surplus was balanced mainly by exopolysaccharide and glycogen biosyntheses at highD values, while cellodextrin excretion occurred mainly at lower ones. With increasing D, the specific rate of carbon flowing down catabolites increased as well, but when expressed as a percentage of carbon it declined, while the percentage of carbon directed through biosynthesis pathways was enhanced. The maximum growth and energetic yields were lower than those obtained in cellulose-limited chemostats and were related to an uncoupling between catabolism and anabolism leading to an excess of energy. Compared to growth on cellobiose in ammonium-limited chemostats (E. Guedon, M. Desvaux, and H. Petitdemange, J. Bacteriol. 182:2010–2017, 2000), (i) a specific consumption rate of carbon of as high as 26.72 mmol of hexose equivalent g of cells−1h−1 could not be reached and (ii) the proportions of carbon directed towards cellodextrin, glycogen, and exopolysaccharide pathways were not as high as first determined on cellobiose. While the use of cellobiose allows highlighting of metabolic limitation and regulation of C. cellulolyticumunder ammonium-limited conditions, some of these events should then rather be interpreted as distortions of the metabolism. Growth of cellulolytic bacteria on easily available carbon and nitrogen sources represents conditions far different from those of the natural lignocellulosic compounds.

Evaluation of the growth and sporulation of different entomopathogenic fungi in different liquid and solid media at varied concentrations

2017 ◽  
Vol 6 (8) ◽  
pp. 5459
Author(s):  
Chandra Teja K. ◽  
Rahman S. J.
Keyword(s):  
Entomopathogenic Fungi ◽  
Large Scale ◽  
Insect Pests ◽  
Culture Media ◽  
Virulent Strain ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Maximum Growth ◽  
Nutritional Requirements ◽  
Virulent Strains

Entomopathogenic fungi like Beauveria bassiana, Metarhizium anisopliae and Lecanicillium lecanii are used in biological control of agricultural insect pests. Their specific mode of action makes them an effective alternative to the chemical Insecticides. Virulent strains of Entomopathogenic fungi are effectively formulated and used as bio-insecticides world-wide. Amenable and economical multiplication of a virulent strain in a large scale is important for them to be useful in the field. Culture media plays a major role in the large-scale multiplication of virulent strains of Entomopathogens. Different substrates and media components are being used for this purpose. Yet, each strain differs in its nutritional requirements for the maximum growth and hence it is necessary to standardize the right components and their optimum concentrations in the culture media for a given strain of Entomopathogen. In the current study, three different nitrogen sources and two different carbon sources were tried to standardize the mass multiplication media for seven test isolates of Entomopathogenic fungi. A study was also conducted to determine the ideal grain media for the optimum conidial yields of the test isolates. Yeast extract was found to be the best Nitrogen source for the isolates. The isolates tested, differed in their nutritional requirements and showed variation in the best nitrogen and carbon sources necessary for their growth. Variation was also found in the optimum concentration of both the ingredients for the growth and sporulation of the isolates. In the solid-state fermentation study, rice was found to be the best grain for the growth of most of the fungi followed by barley. The significance of such a study in the development of an effective Myco-insecticide is vital and can be successfully employed in agriculture is discussed.

Metabolic Flux Analysis and Optimal Control in the Process of Ectoine Fermentation

2011 ◽  
Vol 393-395 ◽  
pp. 851-854
Author(s):  
Lin Hua Zhang ◽  
Xin Zheng ◽  
Ya Jun Lang
Keyword(s):  
Optimal Control ◽  
Batch Fermentation ◽  
Metabolic Flux ◽  
Monosodium Glutamate ◽  
Nitrogen Sources ◽  
Flux Analysis ◽  
Key Factors ◽  
Carbon And Nitrogen Sources ◽  
Carbon And Nitrogen ◽  
Key Nodes

In this study, the metabolic network of ectoine by Halomonas venusta DSM 4743 was established. The key nodes to influence the ectoine fermentation in metabolic flux and the basis during optimal control of fermentation process were investigated. The results showed that G6P, α-KG and OAA nodes were the key factors to influence the synthesis of ectoine. The metabolic flux distributions at the key nodes were significantly improved and ectoine concentration was enhanced in ectoine fermentation by adopting monosodium glutamate as the sole carbon and nitrogen sources, feeding monosodium glutamate and supplying oxygen limitedly. The batch fermentation was carried out in 10 L fermentor , the concentration and yield of ectoine was 8.4 g/L and 0.1 g/g, respectively, which were increased by 2.8 and 2 times, by comparison with batch fermentation using glucose as carbon source.

Amino acid uptake systems in Bacteroides ruminicola

1979 ◽  
Vol 25 (10) ◽  
pp. 1161-1168 ◽  
Author(s):  
Roselynn M. W. Stevenson
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Nitrogen Sources ◽  
Amino Acid Uptake ◽  
Defined Medium ◽  
High Rate ◽  
Transport Systems ◽  
Acid Uptake ◽  
Chemical Structures ◽  
Kinetic Inhibition

Uptake of amino acids by Bacteroides ruminicola was observed in cells grown in a complete defined medium, containing ammonia as the nitrogen source. A high rate of uptake occurred only in fresh medium, as an inhibitory substance, possibly acetate, apparently accumulated during growth. All amino acids except proline were taken up and incorporated into cold trichloroacetic acid precipitable material. Different patterns of incorporation and different responses to 2,4-dinitrophenol and potassium ferricyanide indicated multiple uptake systems were involved. Kinetic inhibition patterns suggested six distinct systems were present for amino acid uptake, with specificities related to the chemical structures of the amino acids. Thus, the failure of free amino acids to act as sole nitrogen sources for growth of B. ruminicola is not due to the absence of transport systems for these compounds.

QUALITATIVE STUDIES OF SOIL MICROORGANISMS: IX. AMINO ACID REQUIREMENTS OF RHIZOSPHERE BACTERIA

1950 ◽  
Vol 28c (1) ◽  
pp. 1-6 ◽  
Author(s):  
R. H. Wallace ◽  
A. G. Lochhead
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Soil Microorganisms ◽  
Maximum Growth ◽  
Defined Medium ◽  
Chemically Defined Medium ◽  
Special Significance ◽  
Specific Amino Acid ◽  
Pronounced Decrease ◽  
Amino Acid Requirements

A study was made of the more specific amino acid requirements of bacteria from the rhizospheres of clover, flax, and wheat plants for which a chemically defined medium containing 23 amino acids provided essentials for maximum growth. Of seven groups of amino acids, the sulphur-containing group (cysteine, methionine, and taurine) was found to be of special significance, the omission of this group resulting in a pronounced decrease in the percentage of organisms able to develop. Further study of organisms dependent upon this group of amino acids for growth showed methionine to be by far the most essential compound. While evident for bacteria from the rhizosphere of all three crops, the effect was more pronounced in the case of clover than with flax or wheat.

341 OXYGEN TENSION AND EGF AFFECT METABOLISM OF IN VITRO-MATURED CUMULUS-ENCLOSED MOUSE OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 278
Author(s):  
K. A. Preis ◽  
G. E. Seidel Jr ◽  
D. K. Gardner
Keyword(s):  
Glucose Uptake ◽  
Oxygen Tension ◽  
Embryo Development ◽  
Metabolic Activity ◽  
Lactate Production ◽  
Defined Medium ◽  
Blastocyst Development ◽  
Developmental Potential ◽  
Exact Test

In vitro maturation of immature oocytes results in limited success in both clinical and research laboratories. Although reduced oxygen concentration is beneficial to embryo development, the optimal concentration for oocyte maturation has yet to be determined. The objective of this study was to determine whether oxygen tension (20% or 5% O2) affects oocyte physiology. Additionally, the effect of epidermal growth factor (EGF) in maturation medium on oocyte metabolic activity and subsequent embryo development was determined. Cumulus–oocyte complexes (COCs; n = 231) were collected from 28-day-old unprimed F1 (C57BL/6 × CBA/ca) mice. COCs were individually matured in defined medium at 37°C in 6% CO2 in one of four groups (Table 1). For the metabolism study, COCs were further divided into two groups: individual maturation in a 2-µL drop of medium for 16 h (n = 131); or individual maturation in 5-μL for 12 h and then placed in a 0.5-μL drop of medium for 4 h (n = 100), the time of greatest metabolic activity of the COC. At 17 h of maturation, COCs were individually fertilized, and zygotes were individually cultured until 96 h, at which time blastocyst development was assessed. Metabolic profiles were analyzed by ANOVA, and blastocyst rates were analyzed by Fisher's exact test. Maturation rates and blastocyst development were not different between groups. However, at 12–16 h of maturation, metabolism of COCs was affected by both oxygen tension and EGF (Table 1). Concerning metabolism over the entire course of maturation, glucose uptake and lactate production were higher in COCs in 5% O2 + 100 ng EGF (P < 0.05) than in the remaining three groups. There was no difference between 5% O2 and 20% O2 + 100 ng EGF, but 20% O2 caused less glucose uptake and lactate production than did the other three treatment groups (P < 0.05). Results of this study are the first to show that oxygen tension alters COC metabolism: COCs matured under 5% O2 were more active metabolically than COCs matured under 20% O2. The effect of oxygen tension is to some extent moderated by the presence of EGF, as metabolic activity of COCs matured under 20% O2 + 100 ng EGF was closer to that of COCs matured under 5% O2 conditions. Although blastocyst rates were similar across the four groups, embryos derived from oocytes matured in different oxygen tensions may exhibit different developmental potential. In conclusion, results of this study have implications for the improvement of maturation conditions in both clinical and research laboratories. Table 1. Carbohydrate metabolism of individual COCs at 12–16 h of maturation

scholarly journals Cellulose Catabolism by Clostridium cellulolyticum Growing in Batch Culture on Defined Medium

2000 ◽  
Vol 66 (6) ◽  
pp. 2461-2470 ◽  
Author(s):  
Mickaël Desvaux ◽  
Emmanuel Guedon ◽  
Henri Petitdemange
Keyword(s):  
Batch Culture ◽  
Cellulose Degradation ◽  
Growth Arrest ◽  
Dry Weight ◽  
Defined Medium ◽  
Carbon Flow ◽  
Natural Ecosystem ◽  
Clostridium Cellulolyticum ◽  
End Products ◽  
Cellulose Concentration

ABSTRACT A reinvestigation of cellulose degradation by Clostridium cellulolyticum in a bioreactor with pH control of the batch culture and using a defined medium was performed. Depending on cellulose concentration, the carbon flow distribution was affected, showing the high flexibility of the metabolism. With less than 6.7 g of cellulose liter−1, acetate, ethanol, H2, and CO2 were the main end products of the fermentation and cellulose degradation reached more than 85% in 5 days. The electron flow from the glycolysis was balanced by the production of H2 and ethanol, the latter increasing with increasing initial cellulose concentration. From 6.7 to 29.1 g of cellulose liter−1, the percentage of cellulose degradation declined; most of the cellulase activity remained on the cellulose fibers, the maximum cell density leveled off, and the carbon flow was reoriented from ethanol to acetate. In addition to that of previously indicated end products, lactate production rose, and, surprisingly enough, pyruvate overflow occurred. Concomitantly the molar growth yield and the energetic yield of the biomass decreased. Growth arrest may be linked to sufficiently high carbon flow, leading to the accumulation of an intracellular inhibitory compound(s), as observed on cellobiose (E. Guedon, M. Desvaux, S. Payot, and H. Petitdemange, Microbiology 145:1831–1838, 1999). These results indicated that bacterial metabolism exhibited on cellobiose was distorted compared to that exhibited on a substrate more closely related to the natural ecosystem of C. cellulolyticum. To overcome growth arrest and to improve degradation at high cellulose concentrations (29.1 g liter−1), a reinoculation mode was evaluated. This procedure resulted in an increase in the maximum dry weight of cells (2,175 mg liter−1), cellulose solubilization (95%), and end product concentrations compared to a classical batch fermentation with a final dry weight of cells of 580 mg liter−1 and 45% cellulose degradation within 18 days.

scholarly journals Metabolic markers of developmental competence for in vitro-matured mouse oocytes

Reproduction ◽  
2005 ◽  
Vol 130 (4) ◽  
pp. 475-483 ◽  
Author(s):  
Kimberly A Preis ◽  
George Seidel ◽  
David K Gardner
Keyword(s):  
Lactate Production ◽  
Glucose Consumption ◽  
Cumulus Cells ◽  
Defined Medium ◽  
Developmental Competence ◽  
Metabolic Markers ◽  
Oocyte Competence ◽  
Carbohydrate Consumption ◽  
Mouse Oocytes

In vitro maturation of oocytes has enormous potential in assisted reproductive technology, but its use has been limited due to insufficient knowledge of oocyte physiology during this dynamic period and lack of an adequate maturation system. The aim of this study was to characterize the metabolic profiles of three groups of oocytes throughout maturation: cumulus–oocyte complexes (COCs), denuded oocytes, and denuded oocytes co-cultured with cumulus cells. Mouse oocytes were collected from 28-day-old unstimulated females and matured in a defined medium. Oocytes were matured individually and transferred into fresh 0.5 μl drops of medium at 4 h intervals until 16 h. Ultramicrofluorimetry was used to quantitate carbohydrate consumption from and metabolite release into the medium. Glucose consumption and lactate production of COCs increased (P < 0.001) over the maturation interval (0–16 h). Glucose consumption by COCs that subsequently fertilized was higher between 8–12 h of maturation than by COCs that did not fertilize (38 versus 29 pmol/COC per h, respectively; P < 0.01). Lactate production by COCs that subsequently fertilized was higher between 8–16 h of maturation, than by oocytes that did not fertilize (8–12 h, 66 versus 46 pmol/COC per h, P < 0.01; 12–16 h, 56 versus 40 pmol/COC per h, respectively; P < 0.05). These data indicate that the final hours of maturation may hold a unique marker of oocyte competence, as during this time fertilizable COCs take up more glucose and produce more lactate than those not subsequently fertilized.

scholarly journals Lovastatin Biosynthesis by Aspergillus terreus in a Chemically Defined Medium

2001 ◽  
Vol 67 (6) ◽  
pp. 2596-2602 ◽  
Author(s):  
Hassan Hajjaj ◽  
Peter Niederberger ◽  
Philippe Duboc
Keyword(s):  
Carbon Source ◽  
Aspergillus Terreus ◽  
Glucose Repression ◽  
Nitrogen Sources ◽  
Glucose Consumption ◽  
Defined Medium ◽  
Specific Productivity ◽  
Chemically Defined Medium ◽  
Carbon And Nitrogen Sources ◽  
Carbon And Nitrogen

ABSTRACT Lovastatin is a secondary metabolite produced by Aspergillus terreus. A chemically defined medium was developed in order to investigate the influence of carbon and nitrogen sources on lovastatin biosynthesis. Among several organic and inorganic defined nitrogen sources metabolized by A. terreus, glutamate and histidine gave the highest lovastatin biosynthesis level. For cultures on glucose and glutamate, lovastatin synthesis initiated when glucose consumption levelled off. When A. terreus was grown on lactose, lovastatin production initiated in the presence of residual lactose. Experimental results showed that carbon source starvation is required in addition to relief of glucose repression, while glutamate did not repress biosynthesis. A threefold-higher specific productivity was found with the defined medium on glucose and glutamate, compared to growth on complex medium with glucose, peptonized milk, and yeast extract.

Nitrogen nutrition and regulation of cephalosporin production in Streptomyces clavuligerus

1979 ◽  
Vol 25 (1) ◽  
pp. 61-67 ◽  
Author(s):  
Yair Aharonowitz ◽  
Arnold L. Demain
Keyword(s):  
Amino Acids ◽  
Growth Rate ◽  
Nitrogen Nutrition ◽  
Antibiotic Production ◽  
Inorganic Nitrogen ◽  
Amino Nitrogen ◽  
Nitrogen Sources ◽  
Streptomyces Clavuligerus ◽  
Defined Medium ◽  
Repressive Effect

When used as sole nitrogen source, certain amino acids (e.g., proline, asparagine) supported both growth and sporulation by Streptomyces clavuligerus streaked onto solid defined medium. Ammonium supported growth but suppressed sporulation. Amino nitrogen was best for cephalosporin production in liquid defined medium, although urea was almost as useful. A comparison of amino acids showed asparagine and glutamine to be the best nitrogen sources and arginine to be almost as good. Ammonium salts supported a somewhat lower growth rate than asparagine, but antibiotic production was very poor on these inorganic nitrogen sources. Addition of ammonium to asparagine did not affect growth rate but increased mycelial mass; cephalosporin production was reduced by about 75%. Antibiotic production was more closely associated with growth in the absence of ammonium than in its presence, indicating a strong inhibitory and (or) repressive effect of NH4+ on antibiotic production. Ammonium exerted its negative effect when added at 24 h or earlier, i.e. before antibiotic formation began.

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