Effect of calcium carbonate on conversion of starch to ethanol by Schwanniomyces castellii

Conversion of starch to ethanol without its prior hydrolysis is of special interest in the treatment of the starch containing waste-waters. In this work the effect of calcium carbonate on the growth and ethanol production from soluble starch by Schwanniomyces castellii NRC 2676 was studied. When the medium was supplemented with 0.1% calcium carbonate the biomass yield and the total starch consumption were increased by 20% and 22%, respectively, while the ethanol yield decreased for 28% in comparison with the control. The rate of biomass production, measured for the period of time between 0 and 24 h, was the highest in the medium with 0.06% calcium carbonate while the rates of ethanol production and starch consumption decreased with and increase in the concentration of that salt; the media with 0.5% and 0.7% calcium carbonate had 3.5 and 5.5 times lower ethanol production rates, and 2.3 and 3.8 times lower rates of starch consumption than the control medium which was not supplemented with calcium carbonate, respectively. The obtained results also showed that the percentage of starch consumed was higher in the media supplemented with calcium carbonate than in the control. Empirical mathematical expressions are given for the relationship of the biomass and ethanol yields to the concentration of calcium carbonate in the medium.     

Effects of adding calcium and sodium salts to field soils on the incidence of clubroot

The effect on clubroot of the addition to a mineral soil of calcium carbonate, calcium sulphate and sodium carbonate was studied in two field experiments during a 3-yr period. Applications of calcium carbonate (ground limestone) at 10 or 20 t/ha increased the soil pH from 6×7 to 7×9; sodium carbonate at 7×5 t/ha to pH 8×3 and calcium sulphate (gypsum) at 20 t/ha caused a slight depression in pH to 6×6. 98% of cabbage plants showed clubroot symptoms in the untreated plots after 3 yr and the percentages were 0×7, 1×6 and 66×6 for the calcium carbonate, sodium carbonate and calcium sulphate treatments respectively. Yields were significantly increased by all three materials. There were no significant differences in disease incidence or in yield when calcium carbonate was used at 10 or 20 t/ha either applied as a single application or as two half-rate applications. Soil from the calcium carbonate treated plots was used in a greenhouse experiment where the addition of inoculum of Plasmodiophora brassicae resulted in a large increase in clubroot incidence. There was no evidence that a biological suppressor was present in the high pH, low disease incidence soils.     

Urease activity in soils

Summary The availability of Ca from different levels of gypsum and calcium carbonate in a non-saline sodic soil has been investigated. Different levels of tagged gypsum (Ca⁴⁵SO₄.2H₂O) and calcium carbonate (Ca⁴⁵CO₃) (i.e. 0, 25, 50, 75, and 100 per cent of gypsum requirement) were mixed thoroughly in 3.5 Kg of a non-saline alkali soil (ESP, 48.4; ECe, 2.68 millimhos/cm). Dhaincha (Sesbania aculeata) — a legume and barley (Hordeum vulgare L.) — a cereal were taken as test crops. Increasing levels of gypsum caused a gradual increase in the yield of dry matter, content of Ca and K in the plant tops and Ca:Na and (Ca+Mg):(Na+K) ratios in both the crops. Application of calcium carbonate caused a slight increase in the dry matter yield, content of Ca and Mg and Ca:Na and (Ca+Mg):(Na+K) ratios in barley, however, in case of dhaincha there was no such effect. Gypsum application caused a gradual decrease in the content of Na and P in both the crops. Total uptake of Ca, Mg, K, N and P per pot increased in response to gypsum application. The effect of calcium carbonate application on the total uptake of these elements was much smaller on dhaincha, but in barley there was some increasing trend.Increasing application of tagged gypsum and calcium carbonate caused a gradual increase in the concentration and per cent contribution of source Ca in both the crops, although, the rate of increase was considerably more in dhaincha. The availability of Ca from applied gypsum was considerably more than that from applied calcium carbonate. Efficiency of dhaincha to utilize Ca from applied sources was considerably more (i.e. about five times) than that of barley     

Direct fermentation of corn to L(+)-lactic acid byRhizopus oryzae

Summary Rhizopus oryzae NRRL 395 was found capable of fermenting ground corn directly to L(+)-lactic acid in the presence of calcium carbonate. The average yield of L(+)-lactic acid was more than 44% on the basis of the amount of total carbohydrate as glucose consumed.     

Effect of calcium on the performance and morphology ofZymomonas mobilis ATCC 29191 in continuous ethanol fermentations

Summary The effect of calcium chloride, over a concentration range of 0–40mM (equivalent to 0–5.88g/L), on various steady-state parameters associated with the chemostat culture ofZ. mobilis ATCC 29191, in a defined salts medium containing 100g/L glucose, was examined. In contradiction of reports by other authors that calcium chloride (>2g/L) inhibits both the growth rate and ethanol yield while at the same time increasing the biomass yield, it was observed in the present study that 25mM (3.7g/L) CaCl₂ did not appreciably affect any of these fermentation parameters. The D_max (dilution rate for 95% substrate utilization) of 0.22/hr was not affected until the concentration of calcium chloride exceeded 30mM (4.41g/L). The apparent increase in biomass yield produced by calcium can be attributed to the formation of insoluble calcium carbonate. Calcium induced morphological changes of the biomass with filamentous growth were seen at concentrations >30 mM. Sparging with nitrogen gas reduces the dissolved carbon, dioxide, lessens the apparent inhibitory effect of calcium on the ethanol productivity and decreases the degree of filament formation.     

Biomineralization Consolidation of Fresco Wall Paintings Samples by Bacillus sphaericus

Bacterial-induced mineralization has been explored for protection and consolidation of degraded limestone, concrete and plaster by precipitation of calcium carbonate. It is the first time that Bacillus sphaericus was used for consolidating the nonsterilized decayed wall paintings samples by immersing them in sterile nutritional media. The B. sphaericus used in this study produced urease, which catalyzes the hydrolysis of urea (CO(NH₂)₂) into ammonium (NH₄) and carbonate (CO₃^?2) leading to the precipitation of calcium carbonate. The effect of B. sphaericus on wall paintings was determined by recording the evolution of culture media chemistry and examining the treated wall paintings under a scanning electron microscope to show the structural and morphological evolution of calcium carbonate that was investigated in wall paintings models.     

Microbial degradation of formaldehyde in white mineral dispersions preserved with formaldehyde-releasing biocides

Two different formaldehyde-degrading microorganisms, Pseudomonas putida and Methylobacterium extorquens, were isolated from calcium carbonate slurry containing the formaldehyde-releasing biocide (ethylenedioxy) dimethanol. Their relative formaldehyde biodegradation and formic acid production kinetics were studied in broth and in calcium carbonate slurry for each microorganism individually, as well as in mixed cultures. Furthermore, the minimal inhibition concentration (MIC) was determined. The results indicated that in slurry, M. extorquens is more tolerant of formaldehyde than P. putida. In slurry, microbial-induced oxidation of formaldehyde caused a temporary accumulation of formic acid, which is presumed to be responsible for pH drop and destabilisation of the calcium carbonate slurry suspension systems. In addition, the residual formaldehyde concentration was observed to drive dominance and recovery of individual formaldehyde-resistant microorganisms in the slurry. Overall, this investigation indicated that biodegradation of formaldehyde in calcium carbonate slurry is brought about by alternating dominance of bacterial genera of mixed formaldehyde-resistant microbial populations.     

Production of l-lactic acid with immobilized Lactobacillus delbrueckii

Summary Living Lactobacillus delbrueckii cells were entrapped in calcium alginate gel beads and employed both in recycle batch and continuous column reactors to produce l-lactic acid from glucose. The substrate contained l% (w/v) yeast extract as nutrient and 4.8% (w/v) solid calcium carbonate as buffer. The maxiumum lactic acid yield obtained was 97%, of which more than 90% was l-lactic acid. The biocatalyst activity half-life in continuous operation was about 100 d, and only about 10% of the activity was lost during intermittent storage of the bioreactor at +7°C for about 5 months.     

Optimization of Bifidobacterium longum growth by use of calcium carbonate-alginate beads

High concentrations of ammonium and sodium ions inhibited Bifidobacterium longum growth more than a high calcium ion concentration. The optimal pH for B. longum growth was determined to be 5.0 due to the lower accumulation of ammonium ion. To reduce the accumulation of ammonium ion and obtain an enhanced growth of B. longum, the pH of the culture containing immobilized calcium carbonate beads was controlled to 5.0 with ammonia water. The concentrations of ammonium, sodium, and calcium ions in the culture were maintained at the desired level. The maximum cell mass increased to 16.8 g/l, 1.23 times higher than cultures without calcium carbonate beads. The number of viable cells in the culture increased to 5.0 × 10¹⁰, 1.67 times more than cultures without calcium carbonate beads.     

Altered free calcium transients in pig kidney cells (LLC-PK1) cultured with penicillin/streptomycin

Summary The effect of a conventional antibiotic (penicillin/streptomycin) mixture on the widely used kidney epithelial cell line, LLC-PK₁, was investigated by measuring growth and intracellular free calcium. Free calcium concentration was the same in cells cultured for 3 to 7 wk with (“plus”) and without (“minus”) antibiotics both at rest and when challenged with high (14 mM) external calcium. When exposed to vasopressin, minus cells exhibited significantly smaller calcium transients than plus cells. A similar difference existed for transients elicited by a calcium ionophore, 4-br-A23187. After longer periods of culture (>20 wk), minus cells grew slower than plus cells but on reaching confluence (minus cells took 1 day longer) the morphologies and viabilities were indistinguishable. The finding that culture with penicillin/streptomycin reversibly modified some properties of LLC-PK₁ cells, at least partly through altered calcium homeostasis, is of importance for workers using this cell model to study drug effects and raises the general possibility of similar effects on other cultured cells.     

Predicting Biogeochemical Calcium Precipitation in Landfill Leachate Collection Systems

Clogging of leachate collection systems within municipal solid waste landfills can result in greater potential for contaminants to breach the landfill barrier system. The primary cause of clogging is calcium carbonate (CaCO_3(s)) precipitation from leachate and its accumulation within the pore space of the drainage medium. CaCO_3(s) precipitation is caused by the anaerobic fermentation of volatile fatty acids (VFAs), which adds carbonate to and raises the pH of the leachate. An important relationship in modeling clogging in leachate collections systems is a yield coefficient that relates microbial fermentation of VFAs to precipitation of calcium carbonate. This paper develops a new, mechanistically based yield coefficient, called the carbonic acid yield coefficient (Y_H), which relates the carbonic acid (H₂CO₃) produced from microbial fermentation of acetate, propionate, and butyrate to calcium precipitation. The empirical values of Y_H were computed from the changes in acetate, propionate, butyrate, and calcium concentrations in leachate as it permeated through gravel-size material. The theoretical and empirical results show that the primary driver of CaCO_3(s) precipitation is acetate fermentation. Additionally, other non-calcium cations (e.g., iron and magnesium) precipitated with carbonate (CO_2-) when present in the leachate. A common yield between total cations bound to CO₃ ^2- and H₂CO₃ produced, called the calcium carbonate yield coefficient (Y_c), can reconcile the empirical yield coefficient for synthetic and actual leachates.     

Isolation and metagenomic characterization of bacteria associated with calcium carbonate and struvite precipitation in a pure moving bed biofilm reactor-membrane bioreactor

A bench-scale pure moving bed bioreactor-membrane bioreactor (MBBR-MBR) used for the treatment of urban wastewater was analyzed for the identification of bacterial strains with the potential capacity for calcium carbonate and struvite biomineral formation. Isolation of mineral-forming strains on calcium carbonate and struvite media revealed six major colonies with a carbonate or struvite precipitation capacity in the biofouling on the membrane surface and showed that heterotrophic bacteria with the ability to precipitate calcium carbonate and struvite constituted ~7.5% of the total platable bacteria. These belonged to the genera Lysinibacillus, Trichococcus, Comamomas and Bacillus. Pyrosequencing analysis of the microbial communities in the suspended cells and membrane biofouling showed a high degree of similarity in all the samples collected with respect to bacterial assemblage. The study of operational taxonomic units (OTUs) identified through pyrosequencing suggested that ~21% of the total bacterial community identified in the biofouling could potentially form calcium carbonate or struvite crystals in the pure MBBR-MBR system used for the treatment of urban wastewater.     

Study of 2,3-butanediol formation by serratiamarcescens

Summary Serratia marcescens can be used for the fermentation of sugar for the formation of 2,3-butanediol. Presence of 1% calcium carbonate increases the formation of the diol and 0.292% of phosphate gives the maximum percentage yield of the diol. The optimumph for the formation of the diol has been found to be 7.     

Formate Oxidation-Driven Calcium Carbonate Precipitation by Methylocystis parvus OBBP

Microbially induced carbonate precipitation (MICP) applied in the construction industry poses several disadvantages such as ammonia release to the air and nitric acid production. An alternative MICP from calcium formate by Methylocystis parvus OBBP is presented here to overcome these disadvantages. To induce calcium carbonate precipitation, M. parvus was incubated at different calcium formate concentrations and starting culture densities. Up to 91.4% ± 1.6% of the initial calcium was precipitated in the methane-amended cultures compared to 35.1% ± 11.9% when methane was not added. Because the bacteria could only utilize methane for growth, higher culture densities and subsequently calcium removals were exhibited in the cultures when methane was added. A higher calcium carbonate precipitate yield was obtained when higher culture densities were used but not necessarily when more calcium formate was added. This was mainly due to salt inhibition of the bacterial activity at a high calcium formate concentration. A maximum 0.67 ± 0.03 g of CaCO₃ g of Ca(CHOOH)₂⁻¹ calcium carbonate precipitate yield was obtained when a culture of 10⁹ cells ml⁻¹ and 5 g of calcium formate liter⁻¹ were used. Compared to the current strategy employing biogenic urea degradation as the basis for MICP, our approach presents significant improvements in the environmental sustainability of the application in the construction industry.     

Weathering of calcite,pyrite, and sulfur by Thermothrix thiopara in a thermal spring

The mineral weathering capabilities of Thermothrix thiopara were investigated by scanning electron microscopy and energy dispersive X‐ray analysis. Thermothrix thiopara is an extremely thermophilic, sulfur‐oxidizing bacterium that grows in a thermal spring whose principal minerals are calcium carbonate, pyrite, and sulfur. Crystals of these minerals were incubated in situ for periods up to eight days, removed, and examined. Results indicated that T. thiopara is partially responsible for weathering calcium carbonate by the production of sulfuric acid, thereby contributing to the formation of a porous tufa mound. Examination of ultravioletirradiated control crystals indicated that the sulfuric acid produced by T. thiopara caused solubilization of calcium carbonate even in the absence of direct bacterial colonization. Pyrite and sulfur were not visibly weathered, but instead were coated with calcium carbonate precipitate. During eight days incubation, growth forms of T. thiopara colonizing the minerals progressed from unicells to filaments to nets of filaments. Bacteria other than T. thiopara appeared after eight days, indicating an increased diversity.     

Surface-sterilization of Glomus mosseae sporocarps for studying endomycorrhization in vitro

 The effects of sterilization time, sterilizing agents (ethanol, Chloramine T, calcium hypochlorite) and antibiotics (streptomycin and gentamycin) on Glomus mosseae (BEG 12) sporocarp germination and contamination were evaluated. Incubation for 10 s in 96 % ethanol, followed by 10 min in a solution of 2% Chloramine T, 0.02% streptomycin, 0.01% gentamycin and Tween 20, and then 6 min in 6% calcium hypochlorite greatly reduced fungal and bacterial contamination from sporocarps and caused little change in germination rate in water agar medium. Accepted: 4 March 1999     

Mechanisms of the inhibition of enzymatic hydrolysis of waste pulp fibers by calcium carbonate and the influence of nonionic surfactant for mitigation

Recycled paper mills produce large quantities of fibrous rejects and fines which are usually sent to landfills as solid waste. These cellulosic materials can be enzymatically hydrolyzed into sugars for the production of biofuels and biomaterials. Paper mill wastes also contain large amounts of calcium carbonate which inhibits cellulase activity. The calcium carbonate (30%, w/w) decreased 40–60% of sugar yield of unbleached softwood kraft pulp. The prime mechanisms for this are by pH variation, competitive and non-productive binding, and aggregation effect. Addition of acetic acid (pH adjustment) increased the sugar production from 19 to 22 g/L of paper mill waste fibers. Strong affinity of enzyme—calcium carbonate decreased free enzyme in solution and hindered sugar production. Electrostatic and hydrogen bond interactions are mainly possible mechanism of enzyme—calcium carbonate adsorption. The application of the nonionic surfactant Tween 80 alleviated the non-productive binding of enzyme with the higher affinity on calcium carbonate. Dissociated calcium ion also inhibited the hydrolysis by aggregation of enzyme.

     

Production of a New Acidic Polysaccharide,Succinoglucan by Alcaligenes faecalis var. myxogenes

A slimy non-spore-forming bacterium strain 10C3 isolated from soil was motile with peritrichous flagella and named Alcaligenes faecalis var. myxogenes. Studies were made on the conditions necessary for maximal production of a new acidic succinoglucan polysaccharide by this strain in shaken cultures. Much production was observed with sucrose, glucose, xylose, galactose, cellobiose, maltose, fructose, mannose and rhamnose. The yield was greatest with sucrose and decreased in order with the above sugars from about 36 to 23 per cent. The most suitable medium contained 4 per cent sugar, 0.5 per cent yeast extract and one per cent calcium carbonate in tap water. The optimum temperature was 28°C.     

酪氨酸酚裂解酶的固定化与转化条件的优化

以欧文氏菌(Erwinia herbicola)来源的酪氨酸酚裂解酶的重组大肠埃希菌Escherichia coli BL21为研究对象,研究固定化大肠埃希菌生产L-酪氨酸的条件。以海藻酸钠为载体,采用单因素实验分别考察了载体材料、明胶浓度、反应时间、苯酚浓度和辅助剂(二氧化硅、硅藻土和碳酸钙)等因素对L-酪氨酸生产的影响,发现明胶浓度、反应时间、苯酚和碳酸钙等因素的影响较为显著,进而通过正交实验探索最优条件。结果表明,生产L-酪氨酸的最优条件:载体为4%海藻酸钠与6%明胶的混合载体,苯酚浓度0.08 mol/L,反应时间8 h,于载体中添加0.6%碳酸钙。此条件下,连续反应9次后L-酪氨酸的产量达到64.5 g/L,比优化前提高了451.3%。     

Characterization of the exoskeleton of the Antarctic king crab Paralomis birsteini

Ocean acidification is projected to inhibit the biogenic production of calcium carbonate skeletons in marine organisms. Antarctic waters represent a natural environment in which to examine the long‐term effects of carbonate undersaturation on calcification in marine predators. King crabs (Decapoda: Anomura: Lithodidae), which currently inhabit the undersaturated environment of the continental slope off Antarctica, are potential invasives on the Antarctic shelf as oceanic temperatures rise. Here, we describe the chemical, physical, and mechanical properties of the exoskeleton of the deep‐water Antarctic lithodid Paralomis birsteini and compare our measurements with two decapod species from shallow water at lower latitudes, Callinectes sapidus (Brachyura: Portunidae) and Cancer borealis (Brachyura: Cancridae). In Paralomis birsteini, crabs deposit proportionally more calcium carbonate in their predatory chelae than their protective carapaces, compared with the other two crab species. When exoskeleton thickness and microhardness were compared between the chelae and carapace, the magnitude of the difference between these body regions was significantly greater in P. birsteini than in the other species tested. Hence, there appeared to be a greater disparity in P. birsteini in overall investment in calcium carbonate structures among regions of the exoskeleton. The imperatives of prey consumption and predator avoidance may be influencing the deposition of calcium to different parts of the exoskeleton in lithodids living in an environment undersaturated with respect to calcium carbonate.