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[The effects on ethanol fermentation of Saccharomyces cerevisiae by adding Ca2+ and inositol].

In this paper we report the effects of Ca2+ and inositol upon growth, ethanol fermentation and ethanol tolerance of Saccharomyces cerevisiae. The result shows that by adding inositol has notable effects on growth and ethanol tolerance of Saccharomyces cerevisiae (p < 0.05), but has little effects on ethanol fermentation (p = 0.3933). By adding Ca2+ has remarkable effects on ethanol fermentation and ethanol tolerance (p < 0.05), but has little effect on yeast growth (p = 0.1544). By means of adding Ca2+ and inositol has more effects on the growth, ethanol fermentation and ethanol tolerance than the condition of only adding Ca2+ or inositol separately.

Calcium↗

Effect of dolomite, magnesium oxide (MgO) and chalk (CaCO3) on in vitro fermentation of amorphous and crystalline cellulose and meadow hay using inoculum from sheep.

Some minerals can influence some biochemical parameters of rumen fermentation. The objective of this experiment was to determine the effect of different amounts (0.1, 0.25 and 0.5 g) of dolomite and to compare the effect of dolomite, magnesium oxide (MgO) and chalk (CaCO3) upon the end products of rumen fermentation in vitro. Amorphous and crystalline cellulose as well as meadow hay were used as substrates and incubated with buffered rumen fluid in sealed fermentation bottles. In dependence on the amount of dolomite and the kind of substrate an inhibitory effect of dolomite on methane production was evident. Significant differences of methane production were found between the controls, crystalline cellulose and meadow hay with 0.5 g of dolomite. An increase of total gas production was observed for cellulose with both 0.25 and 0.5 g of dolomite and also for meadow hay with 0.5 g of dolomite. It can be concluded that there was a remarkable effect of dolomite on methane production and also a slight effect of magnesium oxide and chalk as compared to the effect of dolomite on the fermentation parameters of incubated substrates.

Animal Feed↗

Ethanol fermentation on glucose/xylose mixture by co-cultivation of restricted glucose catabolite repressed mutants of Pichia stipitis with respiratory deficient mutants of Saccharomyces cerevisiae.

Restricted glucose catabolite repressed mutants of P. stipiti CCY 39501 were selected using UV irradiation. Four mutants were obtained which assimilated glucose slower than the native strain of P. stipitis and the degree of glucose repression was about 2-fold lower for P5-90-133 and P5-200-16 mutants and about 10-fold lower for P5-80-7 and P5-80-35 mutants. P5-80-7 and P5-80-35 produced very small amounts of ethanol from glucose and xylose, whereas P5-90-133 and P5-200-16 fermented sugars at the wild-type level. These two mutants were selected for co-fermentation process with native strain of S. cerevisiae V30 or Ja(a), as well as with their respiratory deficient mutants. During co-culture process of P. stipitis mutants with native strains of S. cerevisiae the ethanol yields obtained ranged from 0.38 to 0.45 g/g, and this alcohol was produced mainly from glucose. But, when also xylose, besides glucose was fermented to ethanol during co-fermentation of both mutant strains, lower yields of ethanol (0.28-0.40 g/g) were obtained.

Coculture Techniques↗

[Advance in the study on liquid fermentation for medicinal fungi].

The fermentation technology of medicinal fungi has been used widely. Improving the condition during the fermentation, getting more product, raising the quality of the product and cutting down the cost can minimize the ratio of input to outcome. People are working hard to achieve the aim. By the alteration of medium and culture condition on Ganoderma lucidum, Grifola frondosa and so on, different fermented products are got, especially secondary metabolires are remarkably different. Ultraviolet is used in selecting high quality Grifola frondosa (Gr 9801), Ganoderma lucidum strain. New high quality strain are selected. With the development of medicinal fungi fermentation, new analysis methods, such as HPCE are developed. The technology can be produced in mass in industry. It has a bright future for its virtues.

Agaricales↗

Estimation for model parameters of batch fermentation kinetics.

Based on the widely accepted mathematical model of fermentation kinetics, an analytical solution is deduced in this paper. To describe the feature of batch fermentation, the parameters of the fermentation kinetics in the analytical solution (i.e., mumax, Ks, beta, YG, YP, and m) are estimated at one strike with POWELL optimization algorithm coded in FORTRAN-77. The experimental data in the example is quoted from a batch lysine fermentation process using Corynebacterium glutamicum. The result shows that: 1) the calculated values of the mathematical model agree very well with the experimental data; 2) the synthesis rate of lysine depends on both the growth rate and the concentration of the biomass.

Algorithms↗

Effects of fermentation on product consistency.

A variety of different fermentation processes has been successfully employed to produce consistent protein-based biopharmaceuticals from genetically engineered animal cells. Chinese hamster ovary (CHO) cells were genetically modified to produce recombinant human soluble CD4, tissue plasminogen activator (tPA) or erythropoietin (EPO). Soluble CD4 was collected from extended perfused fermentations of several months' duration, during which some quantitative loss of DNA copy level, mRNA expression level, and fermentation titer were observed. In one extended run, a novel contaminant appeared in intermediates purified from later harvests. However, in all cases, the final soluble CD4 product was consistent in terms of purity and potency. Evaluation of genetic stability for tPA examined both biological traits at the cellular level as well as potency, purity and structure of product derived from cells at various levels of in vitro age; no significant cell age effects were observed. Similarly, evaluation of the EPO product showed that genetically-determined and process-determined traits such as potency, tryptic peptide mapping, and sialylation were consistent from lot to lot. These data exemplified how process design, process validation, and in-process and quality control assays can be used effectively to ensure the consistency of recombinant products derived from cell culture fermentations.

Animals↗

Ileal effluent as a fermentation substrate: implications for butyrate production in the colon.

Fermentation of fiber can lead to an enhanced production of short chain fatty acids (SCFA) and, hence, contribute to the proposed anticarcinogenic properties of butyrate in the colon. The fermentation of fiber isolates and the corresponding ileal effluents has been compared under in vitro conditions. Yield of SCFA per gram of substrate fermented was similar for isolates and fiber-enriched effluents (approximately 4.9 mmol/g) and it could be inferred that nonfiber components of effluent also generated SCFA. Butyrate production was highest for glucan-based polymers (approximately 30% total SCFA) and, from the measured acidogenic profile, production of SCFA will occur mainly in the proximal colon. The buffering capacity of ileal effluents during fermentation restrict the potential for a reduction in pH during acidogenesis compared to fiber isolates. This buffering capacity could limit the bioavailability of butyrate in the colon and, hence, the ability to satisfy the proposed antineoplastic properties of butyrate in the colon.

Anticarcinogenic Agents↗

[Selection and study of potent lactose-fermenting yeasts].

Whey-fermenting Kluyveromyces cultures were revealed among 105 yeast strains assimilating lactose. Eighteen most potent strains isolated from milk products fermented galactose, sucrose, and raffinose, in addition to lactose. Many yeast strains fermented inulin. Most strains were resistant to cycloheximide and grew in medium containing glucose, NaCl, and ethanol at concentrations of up to 50, 11-12, and 10-12%, respectively (4 degrees C). Three strains had mycocinogenic activity. After fermentation of whey with selected yeast strains at 30 degrees C for 2-3 days, ethanol concentration was 4-5%.

Culture Media↗

Physiological functions of resistant proteins: proteins and peptides regulating large bowel fermentation of indigestible polysaccharide.

Animal studies have shown conclusively that feeding of resistant starch (RS) increases production of large bowel total short-chain fatty acids (SCFAs). However, fermentation products of RS may be affected considerably by other dietary ingredients. In rats fed a 20% high-amylose cornstarch (HAS) with casein as the sole protein source, greater cecal SCFAs production was observed compared with that in rats fed a regular cornstarch diet. However, with this diet, the cecal succinate production was also very high. In contrast, when rice or potato protein with lower digestibility was used in place of casein, cecal succinate production decreased with a concomitant increase in butyrate. These observations suggest that nondigested protein, namely resistant protein, might play a role in correcting an imbalance in the ratio of carbohydrate and nitrogen as fermentative substrates for cecal bacteria and in promoting butyrate production. Epidemiological and biochemical data indicate a possible linkage between the fermentation products of starch (butyrate in particular) and the prevention of colorectal cancer as well as ulcerative colits. Accordingly, a fermentation strategy of RS favoring SCFA production should be established to elucidate the potentially beneficial effects of SCFAs on large bowel physiology.

Amylose↗

The Schizosaccharomyces pombe gene encoding gamma-glutamyl transpeptidase I is regulated by non-fermentable carbon sources and nitrogen starvation.

In our previous study, the first structural gene (GGTI) encoding g-glutamyl transpeptidase was cloned and characterized from the fission yeast Schizosaccharomyces pombe, and its transcription, using the GGTI-lacZ fusion gene, containing the 1,085 bp upstream region from the translational initiation point, was found to be enhanced by sodium nitroprusside and L-buthionine-(S,R)-sulfoximine (BSO). In the present work, regulation of the GGTI gene was further elucidated. Non-fermentable carbon sources, such as acetate and ethanol, markedly enhanced the synthesis of beta-galactosidase from the GGTI-lacZ fusion gene. However, its induction by non-fermentable carbon sources appeared to be independent of the presence of the Pap1 protein. Nitrogen starvation also gave rise to induction of GGTI gene expression in a Pap1-independent manner. The three additional fusion plasmids, carrying 754, 421 and 156 bp regions, were constructed. The sequence responsible for the induction by non-fermentable carbon sources and nitrogen starvation was identified to exist within a -421 bp region of the GGTI gene. Taken together, the S. pombe GGTI gene is regulated by non-fermentable carbon sources and nitrogen starvation.

Artificial Gene Fusion↗

[Influence of Mn2+ on the biotechmycin fermentation].

The effect of Mn2+ on the biotechmycin fermentation by Bioengineered strain WSJ-l-195 was studied. In the fermentation process, Mn2+ could improve the biological potency significantly, especially when Mn2+ concentration was 5 mmol/L added at 24 h. The pH profile of fermentation broth decreased gradually after 5 mmol/L Mn2+ supplemented at 24 h, and PMV was lower than that of the control sample. Further research about the influence of Mn2+ on the biosynthesis of biotechmycin was carried out in the aspect of organic acids. The results showed that concentrations of organic acids in a fermentation with 5 mmol/L Mn2+ supplemented at 24 h had been changed greatly, especially the concentration of propionic acid, of which the highest value was about 6 times as that in the control sample at 84 h. In addition, it was found that the yield of biotechmycin could be improved significantly with tiny amount of propionic acid added. Therefore, it can be concluded that Mn2+ has profound influence on the biosynthesis of biotechmycin: it enriches the biotechmycin precursor pool such as propionic acid and thus improves the yield of biotechmycin.

Culture Media↗

[Fermentation behaviors of recombinant Pichia pastoris under inhibited methanol concentration].

Chemostat culture was performed to characterize the growth, substrate consumption and the hirudin production, and to disclose their interrelations in the fermentation of recombinant Pichia pastoris. The Andrew substrate-inhibited growth model is more suitable than Monod model to simulate the growth of Pichia pastoris on methanol. Therefore, two stationary states can be obtained in the continuous culture at a certain dilution rate because of the substrate inhibition on cell growth. The stationary state could be obtained if only the dilution rate not more than 0.048 h(-1) in the continuous fermentation. The concentrations of cell, methanol and hirudin were constant after 50 h continuous culture with dilution rate at 0.04 h(-1). However, it could not be obtained when the dilution rate more than 0.048 h(-1) because the other stationary point at S > 0.048 h(-1) is unstable. Therefore, it was found that the cell concentration declined and the methanol concentration increased from 2.9 g/L to 18.1 g/L within 18h at dilution rate 0.06 h(-1). Thus, the fed-batch culture with a constant specific growth rate was carried out to disclose the fermentation behavior at high and constant methanol concentration in aid of a methanol sensor. The theoretical maximum specific growth rate, microm = 0.0464 h(-1), was found under critical methanol concentration, Scrit = 3.1 g/L. The growth of P. pastoris was typically methanol-limited at the methanol concentration S < Scrit. It was, however, inhibited at S > Scrit. The maximum specific Hir65 production rate qp was obtained at 0.2 mg/(g x h) when methanol concentration and mu were 0.5 g/L and 0.02 h(-1), respectively. The specific Hir65 production rate qp increased with the increase of mu and S at mu < 0.02 h(-1), and decreased at mu > 0.02 h(-1). The specific methanol consumption rate increased with the increase of S when S < 5 g/L, but decreased when S > 5 g/L. At last, the high Hir65 production rate 0.2 mg/(g x h) was obtained in the fermentation conducted under methanol-limited concentration and mu controlled at 0.5 g/L and 0.02 h(-1), respectively, while the specific methanol consumption rate is low only at 0.04 g/(g x h), showing the potential for the strategy of getting high Hir65 production rate at the low consumption of methanol.

Fermentation↗

[Study on the repeated batch fermentation of lipase by immobilized Rhizopus].

Repeated batch fermentation with Rhizopus arrhizus immobilized by polyurethane was optimized in details including the time to replace, the volume of the replaced medium and the optimal composition of the medium to replace. Immobilized cells showed stability for repeated use. Immobilized cells could be used for 9 batches, lasting 140h in flask and 6 batches in 5L fermenter when the substitute culture medium consisted of (%): soybean flour 3.0, earthnut oil 0.5 and the liquid was replaced by 40% . The time for fermentation was reduced largely and the lipase productivity was 3 - 6 times as high as that in batch fermentation.

Biotechnology↗

[Media optimization for exopolysaccharide by Pholiota squarrosa (Pers. ex Fr.) Quel. AS 5.245 on submerged fermentation].

Our previous work has indicated that mycelium growth and exopolysaccharide accumulation in submerged fermentation by Pholiota squarrosa (Pers. ex Fr.) Quel. AS 5.245 are strongly affected by many internal and external factors, including medium constituents and fermentation conditions. In this study, we use an effective two-phase statistical approach to enhance exopolysaccharide production. In the first phase, Plackett-Burman design was undertaken to evaluate the effects of the twenty factors, i.e., glucose, fructose, maltose, yeast extract, tryptone, K2HPO4, KH2PO4, (NH4)2SO4, NaNO3, FeSO4, MgSO4, MnCl2, ZnCl2, FeCl3, CuSO4.5H2O, vitamin B1, initial pH, the temperature, the medium volume and the duration, to the fermentation. By regression analysis, yeast extract, tryptone, fructose, MgSO4, MnCl2, initial pH and temperature were found to be important for exopolysaccharide production, while glucose, maltose, NaNO3, ZnCl2, vitamin B1, the duration and the volume are important to the mycelium biomass. In the second phase of the optimization process, a response surface methodology (RSM) was used to optimize the above critical internal factors, and to find out the optimal concentration levels and the relationships between these factors. Based on the results of the first phase, a five-level six-factor (yeast extract, fructose, MgSO4, maltose, ZnCl2 and initial pH) central composite rotatable design (CCRD) was employed. By solving the quadratic regression model equation using appropriate statistic methods, the optimal concentrations for obtaining 876.32 microg exopolysaccharide per milliliter of fermentation liquor were calculated as: 6.0g/L yeast extract, 11.5g/L fructose, 0.5g/L MgSO4, 9.6g/L maltose, 38.6mg/L ZnCl2 and with the initial pH 5.3. The experimental data under various conditions have validated the theoretical values.

Culture Media↗

[Continuous ethanol fermentation using self-flocculating yeast in multi-stage suspended bioreactors coupled with directly recycling of waste distillage].

A fermentation system composed of four airlift suspended-bed bioreactors in series and with a total working volume of 4800 mL was established. Continuous ethanol fermentation using self-flocculating yeast SPSC01, a fusant from Saccharomyces cerevisiae and Schizosaccharomyces pombe, and two-stage enzymatic hydrolyte of dry milling corn powder, was continuously run for 120 days. All of the backset distillage collected after distilling the final beer was used to mix the corn powder and no any other wastes except the solid residue of corn powder was discharged from the fermentation system, which guaranteed the distillage to be recycled at its maximum. The experimental results revealed that both ethanol and residual sugar in the final beer could be maintained relatively stable with their average levels of 93.6 and 7.9 g/L, respectively when the fermentation system was operated at the dilution rate of 0.05 h(-1). Parameter oscillations reported previously were also observed for the first and second bioreactors, but were effectively attenuated thereafter, which indicated that high yeast cell concentrations resulted from the self-immobilization of this special self-flocculating strain contributed to damp these oscillations. The monitoring of residual nitrogen and phosphor indicated that the accumulations of these nutritional elements occurred and the amount of these inorganic salts supplemented in the substrate should be decreased properly.

Bioreactors↗

The fermentative production of acetone-butanol by Clostridium acetobutylicum.

Fourteen different media were used in the fermentative production of acetone-butanol. The highest total yields were achieved in medium I. Potato starch and soluble starch were suitable as carbon sources. The best concentrations of potato starch and soluble starch were 500.0 and 10.0 g/l, respectively. Peptone was the most favourable nitrogen source. The best concentration of peptone was 4.0 g/l. Calcium carbonate in 3.6 g/l acted as buffering agent in the fermentation process. The best initial pH value of the fermentation medium was 6.0. The optimum temperature was 32--33degreesC. The fermentation process required 120 h to obtain maximum yields of acetone-butanol.

Acetone↗

Effect of enzymatic pretreatment on solubilization and volatile fatty acid production in fermentation of food waste.

Food waste can be a valuable carbon source in biological nutrient removal (BNR) systems because of the high C/N and C/P ratio. However, pretreatment is necessary to promote hydrolysis of food waste because of the high concentration of volatile solids associated with organic matter. The influence of the enzymatic pretreatment on acid fermentation of food waste was investigated in this study. Solubilization of particulate matter in food waste was carried out using commercial enzymes. The acidification efficiency and the volatile fatty acid (VFA) production potential of enzymatically pretreated food waste were examined. The highest volatile suspended solid (VSS) reduction was obtained with an enzyme mixture ratio of 1:2:1 for carbohydrase: protease: lipase. An optimum enzyme dosage for solubilization of food waste was 0.1% (V/V) with the enzyme mixture ratio of 1:2:1. In the acid fermentation of enzymatically pretreated food waste, the maximum VFA production and the highest VFA fraction in soluble COD (SCOD) were also achieved at 0.1% (V/N) of total enzyme dosage. Increase in VFA production at this level of enzyme dosage was over 300% compared with the control fermenter. The major form of VFA produced by fermentation was n-butyrate followed by acetate.

Acetates↗

Butyrate production from oligofructose fermentation by the human faecal flora: what is the contribution of extracellular acetate and lactate?

Butyrate is an important substrate for maintenance of colonic health and oligofructose fermentation by human faecal bacteria can increase butyrate production in vitro. However, oligofructose appears to be fermented by mainly acetate and lactate-producing bacteria rather than butyrate-producing bacteria. Isotope labelling studies using [U-(13)C(6)]glucose were used to show that (13)C(2) and (13)C(4) were the major labelled butyrate species produced from glucose fermentation, via [(13)C(2)]acetate-acetyl CoA as intermediate. Bacterial interconversion reactions were quantified and acetate conversion to butyrate and lactate conversion to acetate, propionate and butyrate were observed. Addition of oligofructose to faecal batch cultures significantly increased butyrate production. Of the newly synthesised butyrate from oligofructose fermentation, 80 % was derived from interconversion of extracellular acetate and lactate, with acetate being quantitatively more significant. Carbohydrates, such as oligofructose, have prebiotic properties. In addition, oligofructose selectively stimulates the bacterial conversion of acetate and lactate to butyrate. Carbohydrates with similar properties represent a refinement of the prebiotic definition, termed butyrogenic prebiotics, because of their additional functionality.

Acetates↗