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PubMed2002

Solid state fermentation of broiler litter for production of biocontrol agents.

Several varieties of heat-sterilized broiler litter with 60% (wet basis, wb) moisture content were substrate in solid-state fermentations to produce biocontrol agents. Litter varieties included litter produced by one flock of broilers from medicated and non-medicated controlled rations, and litter produced by two flocks and four flocks on a single application of bedding material from medicated commercial sources. Litter preparations were inoculated with monocultures of Bacillus thuringiensis serovar japonensis strain Buibui, a pathogen of Japanese beetle larvae (Popillia japonica), or Pseudomonas fluorescens 2-79. B. thuringiensis did not grow in unextracted 1-flock litter nor in water extracted litter, but grew in methanol extracted litter to 5 x 10(10) cell forming units (CFU)/g litter (dry weight, dw) and a spore count of 1 x 10(10) CFU/g litter (dw). B. thuringiensis also grew in unprocessed 2-flock and 4-flock litter, achieving cell counts of 3 x 10(9) and 1 x 10(9) CFU/g litter (dw), respectively, and spore counts of 1 x 10(9) CFU/g litter (dw). P. fluorescens grew in medicated 1-flock litter with no extraction to a cell density greater than 4 x 10(11) CFU/g litter (dw). Bioassays in soil containing over 0.5% (db) litter fermented with B. thuringiensis resulted in over 90% mortality in 21 days for first instars of Japanese beetle when compared to a control treatment using compost without fermented litter. The investigations demonstrate that bacterial biocontrol agents produced via solid substrate fermentations using broiler poultry litter have potential in biocontrol applications in the soil environment.

Animals↗
PubMed2002

Oenological properties of Hanseniaspora osmophila and Kloeckera corticis from wines produced by spontaneous fermentations of normal and dried grapes.

Strains of Hanseniaspora osmophila and Kloeckera corticis, isolated from wines produced by spontaneous fermentations of normal and dried grapes, were characterized for their fermentation behavior with and without SO(2) at 25 degrees C. All isolates behaved as glucophilic yeasts and yielded ethanol at concentrations of about 9% (v/v); acetic acid, acetaldehyde, ethyl acetate and acetoin were always produced to high concentrations. SO(2) addition had no significant effect on growth yield and fermentation rate. These metabolic features were maintained in the presence of 400 g l(-1) of sugars and at 15 degrees C, and were quite similar to those shown by Saccharomycodes ludwigii. Therefore, H. osmophila and K. corticis should be considered detrimental yeast species, particularly in fermentations of musts from dried grapes.

Ascomycota↗
PubMed2006

[Detection of intracellular reactive oxygen species by flow cytometry in Pichia pastoris fermentation].

In Pichia pastoris fermentation, methanol was oxidized into carbon oxide and produced a byproduct H2 O2, one of the partially reduced forms of molecular oxygen known as reactive oxygen species (ROS) . ROS are highly damaging towards cellular constituents. Flow cytometry (FCM) is an excellent method that permits the rapid, optical analysis of individual cells and has many advantages over conventional cytometry. However, its use in detecting intracellular ROS levels during Pichia fermentation was rarely reported. In our work, by means of flow cytometry, two fluorescent dye 2', 7'-dichlorofluorescin diacetate (DCFH-DA) and propidium iodide (PI) were used to detect ROS. The effect of intracellular ROS on Pichia pastoris cells during fermentation was studied through the comparison between DCFH-DA/PI double-stained cells and PI single-stained cells. In this study, the loss of cell viability during fermentation was correlated with the accumulation of ROS. At the glycerol batch and fed-batch phase, little ROS was accumulated intracellularly and cell viability reached almost 100%. At the early methanol fed-batch phase, intracellular ROS accumulation took place but 98.5% cells still kept viable. At the later methanol fed-batch phase, 94.0% cells accumulated high ROS. As a result, some cells lost their viability because of the damage of ROS. 25.4% dead cells accumulated high ROS in the total 29.1% dead cells.

Bioreactors↗
PubMed2006

[Continuous ethanol fermentation coupled with recycling of yeast flocs].

A continuous ethanol fermentation system composed of three-stage tanks in series coupled with two sedimentation tanks was established. A self-flocculating yeast strain developed by protoplast fusion from Saccharomyces cerevisiae and Schizosaccharomyces pombe was applied. Two-stage enzymatic hydrolysate of corn powder containing 220g/L of reducing sugar, supplemented with 1.5g/L (NH4)2HPO4 and 2.5g/L KH2PO4, was used as the ethanol fermentation substrate and fed into the first fermentor at the dilution rate of 0.057h(-1). The yeast flocs separated by sedimentation were recycled into the first fermentor as two different models: activation-recycle and direct recycle. The quasi-steady states were obtained for both operation models after the fermentation systems experienced short periods of transitions. Activation process helped enhance the performance of ethanol fermentation at the high dilution rates. The broth containing more than 101g/L ethanol, 3.2g/L residual reducing sugar and 7.7g/L residual total sugar was produced. The ethanol productivity was calculated to be 5.77g/(L x h), which increased by more than 70% compared with that achieved in the same tank in series system without recycling of yeast cells.

Biomass↗
PubMed2000

Changes to the quantity and processing of starchy foods in a western diet can increase polysaccharides escaping digestion and improve in vitro fermentation variables.

This study investigated how readily achievable changes to the quantity and processing of starchy foods in a typical Western diet: (1) were reflected in levels of resistant starch (RS) and NSP excreted from the small intestine; and (2) more favourable profiles of butyrate, NH3 and phenol production. Two diets, a low-starch diet (LSD) and a high-starch, low-fat diet (HSLFD) were compared. The LSD with 20% total energy (%E) from starch was based on a 'typical' Australian diet, while the HSLFD (40%E as starch) was the same Australian diet modified by an increased content of legumes, starchy foods and coarsely-ground cereals and by a reduced fat content. Four subjects with iliostomies consumed each diet for 2 d, with ileal effluent collection on the second day. On the HSLFD compared with the LSD, RS in ileal effluent increased from from 0.49 to 1.7 g/MJ per d (P < 0.005) while ileal NSP excretion increased from 2.0 to 3.3 g/MJ per d (P < 0.05). Ileal effluents obtained after each diet were incubated for 24 h in vitro with a human faecal innoculum. After fermentation, ileal effluent from the HSLFD produced more butyrate relative to other short-chain fatty acids (17.5 v. 15.8 molar %, P < 0.005) and less phenol (2.3 v. 5.7 mg/l, P < 0.05) and NH3 (20.3 v. 23.1 mmol/l, P < 0.005) than the LSD diet. The HSLFD also generated a lower pH (6.15 v. 6.27, P < 0.05). On a wt/wt basis, RS was 2.3-fold higher in the HSLFD effluent while NSP did not increase, suggesting that the change in RS largely contributed to the fermentation effects. Changes in in vitro variables when the HSLFD ileal effluent was ground before fermentation indicated the importance of physical structure in determining ileal excretion of RS. We conclude that: (1) readily achievable modifications to the amount and processing of starchy foods in an Australian diet would produce potential benefits for in vitro fermentation variables; and (2) the physical structure of grains and cereals is important in determining access by colonic bacteria to a carbohydrate substrate.

Adolescent↗
PubMed1990

Noninvasive method for monitoring ethanol in fermentation processes using fiber-optic near-infrared spectroscopy.

Short-wavelength near-infrared (SW-near-IR) spectroscopy (700-1100 nm) is used for the determination of ethanol during the time course of a fermentation. Measurements are performed noninvasively by means of a photodiode array spectrometer equipped with a fiber-optic probe placed on the outside of the glass-wall fermentation vessel. Pure ethanol/water and ethanol/yeast/water mixtures are studied to establish the spectral features that characterize ethanol and to show that determination of ethanol is independent of the yeast concentration. Analysis of the second-derivative data is accomplished with multilinear regression (MLR). The standard error of prediction (SEP) of ethanol in ethanol/water solutions is approximately 0.2% over a range of 0-15%; the SEP of ethanol in ethanol/yeast/water solutions is 0.27% (w/w). Results from the mixture experiments are then applied to actual yeast fermentations of glucose to ethanol. By use of a gas chromatographic method for validation, a good correlation is found between the intensity of backscattered light at 905 nm and the actual ethanol. Additional experiments show that a calibration model created for one fermentation can be used to predict ethanol production during the time course of others with a prediction error of 0.4%.

Ethanol↗
PubMedDate not supplied

Dissolved carbon dioxide measurement and its correlation with operating parameters in fermentation processes.

A silicone tubing probe was used to measure dissolved carbon dioxide in two industrial fermentation processes. The two processes differed markedly in their broth viscosities: one had high viscosity (2000-3000 cP) and the other had low viscosity (< 100 cP). The goal was to study changes in dissolved carbon dioxide concentration in fermentation broth when operating parameters, such as total pressure, air flow rate and, agitation rate, were modified. It was shown that in high-viscosity fermentation broths the dissolved carbon dioxide concentration can reach as high as 190% of that calculated by assuming an equilibrium between the air coming out of the fermentor and the fermentation broth. For low-viscosity broth, the highest dissolved carbon dioxide concentration was found to be about 133% of the equilibrium value. On the basis of the material balance over the entire fermentor, the ratio of actual dissolved carbon dioxide partial pressure and carbon dioxide partial pressure in air leaving the fermentor was correlated to the operating parameters of pressure, air flow, and carbon dioxide transfer coefficient. Since carbon dioxide transfer coefficients are not readily available, we recommend a method of using oxygen transfer coefficients to develop a correlation that can be used on-line to predict changes in dissolved carbon dioxide concentration when operating parameters are changed or when process improvements and scale-up are undertaken.

Biotechnology↗
PubMedDate not supplied

Sugar recovery and fermentability of hemicellulose hydrolysates from steam-exploded softwoods containing bark.

The hemicellulose sugar recovery and ethanol production obtained from SO2-catalyzed steam explosion of a mixed white fir (70%) and ponderosa pine (30%) feedstock containing bark (9% dry weight/dry weight) was assessed. More than 90% of the available hemicellulose sugars could be recovered in the hydrolysate obtained after steam explosion at 195 degrees C, 2.38 min, and 3.91% SO2, with 59% of the original hemicellulose sugars detected in a monomeric form. Despite this high sugar recovery, this hydrolysate showed low ethanol yield (64% of theoretical yield) when fermented with a spent sulfite liquor-adapted strain of Saccharomyces cerevisiae. In contrast, most hydrolysates prepared at higher steam explosion severity showed comparable or higher ethanol yields. Furthermore, the hydrolysates prepared from bark-free feedstock showed better fermentability (87% of theoretical yield) despite containing higher concentration of known inhibitors. The ethanol yield from the hydrolysate prepared from a bark-containing wood sample could be improved to 81% by an extra stage acid hydrolysis (121 degrees C for 1 h in 3% sulfuric acid). This extra stage acid hydrolysis and steam explosion at higher severity conditions seem to improve the fermentability of the hydrolysates by transforming certain inhibitory compounds present in the hydrolysates prepared from the bark-containing feedstock and thus lowering their inhibitory effect on the yeast used for the ethanol fermentation.

Abies↗
PubMedDate not supplied

Design and installation of a next generation pilot scale fermentation system.

Four new fermenters were designed and constructed for use in secondary metabolite cultivations, bioconversions, and enzyme production. A new PC/PLC-based control system also was implemented using GE Fanuc PLCs, Genius I/O blocks, and Fix Dynamics SCADA software. These systems were incorporated into an industrial research fermentation pilot plant, designed and constructed in the early 1980s. Details of the design of these new fermenters and the new control system are described and compared with the existing installation for expected effectiveness. In addition, the reasoning behind selection of some of these features has been included. Key to the design was the goal of preserving similarity between the new and previously existing and successfully utilized fermenter hardware and software installations where feasible but implementing improvements where warranted and beneficial. Examples of enhancements include strategic use of Inconel as a material of construction to reduce corrosion, piping layout design for simplified hazardous energy isolation, on-line calculation and control of nutrient feed rates, and the use of field I/O modules located near the vessel to permit low-cost addition of new instrumentation.

Algorithms↗
PubMedDate not supplied

Influence of ambient air temperature on the cooling/heating load of a single cell protein jacketed fermenter operating on cheese whey under continuous conditions.

The heat generated by mixing and lactose metabolism, during the continuous production of single cell protein from cheese whey lactose using a jacketed fermenter with running cooling water, was calculated using a heat balance equation. The technique quantified the heat produced in and lost from the fermentation unit. Most of the heat generated by mixing in the cell-free system (97.47%) was lost with exhaust gas, while a very small amount (2.53%) was lost through the fermenter lid, wall, and bottom. The heat generated by mixing was significant (26.31% of the total heat generated in the fermentation system with an active yeast population present) and, therefore, cannot be ignored in heat balance calculations. About 19.71% of the total heat generated in the reactor was lost through the coolant at an ambient temperature of 22 +/- 0.5 degrees C, showing the need for a cooling system. A yeast population size of 986 million cells/mL and a lactose removal efficiency of 95.6% were observed. About 72.5% and 27.5% of the lactose consumed were used for growth and respiration, respectively. A yield of 0.66 g of cells/g of lactose was achieved. The heat released by unit biomass was 7.05 kJ/g of cells. The results showed the significant impact of ambient air temperature on the cooling load. The heat to be removed from the medium by the cooling system varied from 3.46 to 281.56 kJ/h when the temperature increased from 16 to 30 degrees C. A heating system is needed to maintain the medium temperature at 34 degrees C when the ambient air temperature is below 16 degrees C.

Aerobiosis↗
PubMedDate not supplied

Identification of critical batch operating parameters in fed-batch recombinant E. coli fermentations using decision tree analysis.

To develop a useful fermentation process model, it is first necessary to identify which batch operating parameters are critical in determining the process outcome. To identify critical processing inputs in large databases, we have explored the use of Decision Tree Analysis with the decision metrics of Gain (i.e., Shannon Entropy changes), Gain Ratio, and a multiple hypergeometric distribution. The usefulness of this approach lies in its ability to treat "categorical" variables, which are typical of archived fermentation databases, as well as "continuous" variables. In this work, we demonstrate the use of Decision Tree Analysis for the problem of optimizing recombinant green fluorescent protein production in E. coli. A database of 85 fermentations was generated to examine the effect of 15 process input parameters on final biomass yield, maximum recombinant protein concentration, and productivity. The use of Decision Tree Analysis led to a considerable reduction in the fermentation database through the identification of the significant as well as insignificant inputs. However, different decision metrics selected different inputs and different numbers of inputs to classify the data for each output.

Algorithms↗
PubMedDate not supplied

Design of methanol Feed control in Pichia pastoris fermentations based upon a growth model.

The methylotrophic yeast Pichia pastoris is an effective system for recombinant protein productions that utilizes methanol as an inducer, and also as carbon and energy source for a Mut(+) (methanol utilization plus) strain. Pichia fermentation is conducted in a fed-batch mode to obtain a high cell density for a high productivity. An accurate methanol control is required in the methanol fed-batch phase (induction phase) in the fermentation. A simple "on-off" control strategy is inadequate for precise control of methanol concentrations in the fermentor. In this paper we employed a PID (proportional, integral and derivative) control system for the methanol concentration control and designed the PID controller settings on the basis of a Pichia growth model. The closed-loop system was built with four components: PID controller, methanol feed pump, fermentation process, and methanol sensor. First, modeling and transfer functions for all components were derived, followed by frequency response analysis, a powerful method for calculating the optimal PID parameters K(c) (controller gain), tau(I) (controller integral time constant), and tau(D) (controller derivative time constant). Bode stability criteria were used to develop the stability diagram for evaluating the designed settings during the entire methanol fed-batch phase. Fermentations were conducted using four Pichia strains, each expressing a different protein, to verify the control performance with optimal PID settings. The results showed that the methanol concentration matched the set point very well with only small overshoot when the set point was switched, which indicated that a very good control performance was achieved. The method developed in this paper is robust and can serve as a framework for the design of other PID feedback control systems in biological processes.

Bioreactors↗
PubMedDate not supplied

Solid-state fermentation for production of griseofulvin on rice bran using Penicillium griseofulvum.

Griseofulvin is a secondary metabolite produced from fungal species that have morphology suitable for solid-state fermentation (SSF). Reports on production of griseofulvin by SSF are scarce. The present work investigates SSF for griseofulvin production, optimization of its process parameters vis-à-vis the conventional submerged fermentation and its downstream processing from the same. Rice bran adjusted to an initial moisture content (IMC) of 50% (v/w) inoculated with 1 mL of a suspension of 10(6) spores/mL under agitation at 250 rpm containing the modified Czapek-Dox medium and additional 0.1% choline chloride as a precursor gave a yield of griseofulvin in 9 days that was comparable to submerged fermentation after 28 days. The yield of griseofulvin (microg/g dry biomass) was comparable in SSF and submerged fermentation. The biomass was estimated by estimation of chitin. Discussions on the effect of each parameter in SSF have also been included.

Chromatography, Thin Layer↗
PubMedDate not supplied

Adsorption-desorption process using wood-based activated carbon for recovery of biosurfactant from fermented distillery wastewater.

Methods used for biosurfactant recovery include solvent extraction, precipitation, crystallization, centrifugation and foam fractionation. These methods cannot be used when distillery wastewater (DW) is used as the nutrient medium for biosurfactant production by Pseudomonas aeruginosa strain BS2, because recovery of biosurfactant by any of these methods imparts color to the biosurfactant. The biosurfactant has a nonaesthetic appearance with lowered surface active properties. These methods cannot be used for continuous recovery of biosurfactant during cultivation. Hence, a new downstream technique for biosurfactant recovery from fermented DW comprised of adsorption-desorption processes using wood-based activated carbon (WAC) was developed. This study involves batch experiments to standardize the factors affecting the rate of biosurfactant adsorption onto WAC. WAC was the most efficient adsorbent among various ones tested (i.e., silica gel, activated alumina and zeolite). The WAC (1% w v(-1)), equilibrium time (90 min), pH range of 5-10 and temperature of 40 degrees C were optimum to achieve 99.5% adsorption efficiency. Adsorption kinetics and intraparticle diffusion studies revealed the involvement of both boundary layer diffusion and intraparticle diffusion. The Langmuir adsorption isotherm of WAC indicated the formation of a monolayer coverage of the biosurfactant over a homogeneous carbon surface, while the Freundlich isotherm showed high adsorption at strong solute concentrations and low adsorption at dilute solute concentrations. WAC concentration of 4% w v(-1) facilitated complete removal of the biosurfactant from collapsed foam (contained 5-fold higher concentration of biosurfactant than was present in fermented DW). Biosurfactant adsorption was of chemisorption type. Acetone (polar solvent) was a specific viable eluant screened among various ones tested because it selectively facilitated maximum recovery, i.e., 89% biosurfactant from WAC. By acetone treatment, complete regeneration of WAC was feasible and WAC can be reused for biosurfactant recovery up to 3 cycles. The recovered biosurfactant showed improved surface-active property (i.e., much lower critical micelle concentration value of 0.013 verses 0.028 mg mL(-1) for biosurfactant recovered by classical methods). The reuse potential of WAC was assessed and results suggest that the carbon can be reused for three consecutive cycles for biosurfactant adsorption from fermented wastewater without any decrease in adsorption efficiency. Thus, this process forms a basis for continuous recovery of biosurfactant from fermented DW and concentrated foam. This process reduces the use of high cost solvent, avoids end product inhibition and minimizes product degradation.

Adsorption↗
PubMedDate not supplied

Continuous beer fermentation using immobilized yeast cell bioreactor systems.

Traditional beer fermentation and maturation processes use open fermentation and lager tanks. Although these vessels had previously been considered indispensable, during the past decades they were in many breweries replaced by large production units (cylindroconical tanks). These have proved to be successful, both providing operating advantages and ensuring the quality of the final beer. Another promising contemporary technology, namely, continuous beer fermentation using immobilized brewing yeast, by contrast, has found only a limited number of industrial applications. Continuous fermentation systems based on immobilized cell technology, albeit initially successful, were condemned to failure for several reasons. These include engineering problems (excess biomass and problems with CO(2) removal, optimization of operating conditions, clogging and channeling of the reactor), unbalanced beer flavor (altered cell physiology, cell aging), and unrealized cost advantages (carrier price, complex and unstable operation). However, recent development in reactor design and understanding of immobilized cell physiology, together with application of novel carrier materials, could provide a new stimulus to both research and application of this promising technology.

Beer↗
PubMed2005

Modeling the dynamics of fermentation and respiratory processes in a groundwater plume of phenolic contaminants interpreted from laboratory- to field-scale.

A biodegradation model with consecutive fermentation and respiration processes, developed from microcosm experiments and simulated mathematically with microbial growth kinetics, has been implemented into a field-scale reactive transport model of a groundwater plume of phenolic contaminants. Simulation of the anaerobic plume core with H2 and acetate as intermediate products of biodegradation allows the rates and parameter values forfermentation processes and individual respiratory terminal electron accepting processes (TEAPS) to be estimated using detailed, spatially discrete, hydrochemical field data. The modeling of field-scale plume development includes consideration of microbial acclimatization, substrate toxicity toward degradation, bioavailability of mineral oxides, and adsorption of biogenic Fe(ll) species in the aquifer, identified from complementary laboratory process studies. The results suggest that plume core processes, particularly fermentation and Fe(lll)-reduction, are more important for degradation than previously thought, possibly with a greater impact than plume fringe processes (aerobic respiration, denitrification, and SO4-reduction). The accumulation of acetate as a fermentation product within the plume contributes significantly to the mass balance for carbon. These results demonstrate the value of quantifying fermentation products within organic contaminant plumes and strongly suggest that the conceptual model selected for reactive processes plays a dominant role in the quantitative assessment of risk reduction by naturally occurring biodegradation processes.

Acetates↗
PubMed2000

Saccharomyces cerevisiae mannoproteins that protect wine from protein haze: their release during fermentation and lees contact and a proposal for their mechanism of action.

A fraction containing the mannoproteins released during fermentation from the winemaking strain of Saccharomyces cerevisiae, Maurivin PDM, was able to reduce the visible protein haze in white wine. This fraction of haze protective mannoprotein material (HPM) could be recovered by either ultrafiltration or ethanol precipitation. The kinetics of the release of both mannose- and glucose-containing polymers during the growth cycle of PDM were determined as a guide to the release of HPM. Active HPM was first detected in the culture supernatant when the cells were exponentially growing. HPM was also released into the medium under an environment simulating winemaking conditions by PDM cells during fermentation as well as during storage on yeast lees. Since the amounts of HPM released during fermentation are greater than those subsequently extracted from the cell wall, fermentation would be a more viable procedure than extraction from yeast cells for the commercial production of HPM. Yeast invertase, a mannoprotein with haze protective activity, was used as a model substrate to investigate the mechanism of haze protection. Invertase was found to reduce visible turbidity but not prevent protein precipitation. Invertase itself did not precipitate but remained soluble in the wine. On the basis of these observations, we propose that the mechanism of haze protection may be one of competition between HPM and wine proteins for unknown wine component(s), the latter being required for the formation of large insoluble aggregates of denatured protein. As the available concentration of these components decreases, due to the presence of HPM, the particle size of the haze decreases and thus visible turbidity declines.

Fermentation↗
PubMed2001

Monitoring ester formation in grape juice fermentations using solid phase microextraction coupled with gas chromatography-mass spectrometry.

Volatile esters contribute important floral and fruity sensory properties to wine. Numerous factors influence the biosynthesis and hydrolysis of esters throughout yeast fermentation; however, methods to monitor the dynamic changes in ester production that occur during winemaking processes are limited. In this study, we showed that solid phase microextraction (SPME), a rapid, solventless sampling procedure, combined with GC/MS analysis is a useful method for the nearly continuous analysis of volatile compounds such as esters that are produced during fermentation. Accuracy, precision, and limits of quantification were comparable to those of other sample preparation methods such as liquid-liquid extraction. Using GC/MS-SPME to monitor fatty acid ethyl esters and acetate esters, we obtained detailed information on the production patterns of ester formation during fermentation. This method now enables the monitoring of volatiles during fermentation and can provide greater insight into yeast metabolism and flavor formation.

Beverages↗