Chitosan/albumin/CaCO3 as mimics for membrane bioreactor fouling: genesis of structural mineralized-EPS-building blocks and cake layer compressibility

Membrane bioreactor biofouling is usually described as an extracellular matrix in which biopolymers, inorganic salts and active microbes co-exist. For that reason, biomineralization (BM) models can be useful to describe the spatial organization and environmental constraints within the referred scenario. BM arguments were utilized as background in order to (1) evaluate CaCO₃ influence on flux decline; pore blocking and cake layer properties (resistance, permeability and compressibility) in a wide range of Chitosan/Bovine serum albumin (BSA) mixtures during step-pressure runs and, (2) perform membrane autopsies in order to explore the genesis of mineralized extracellular building blocks (MEBB) during cake layer build up. Using low molecular weight chitosan (LC) and BSA, 2 L of 5 LC/BSA mixtures (0.25-1.85 ratio) were pumped to an external ultra filtration (UF) membrane (23.5 cm², hydrophobic, piezoelectric, 100 kDa as molecular weight cut-off). Eight different pressure steps (40 ± 7 to 540 ± 21 kPa) were applied. Each pressure step was held for 900 s. CaCO₃ was added to LC/BSA mixtures at 0.5, 1.5 and 3 mM in order to create MEBB during the filtration tests. Membrane autopsies were performed after the filtration tests using thermo gravimetric, scanning microscopy and specific membrane mass (mg cm⁻²) analyses. Biopolymer-CaCO₃ step-pressure filtration created compressible cake layers (with inner voids). The formation of an internal skeleton of MEBB may contribute to irreversible fouling consolidation. A hypothesis for MEBB genesis and development was set forth.     

Arsenic and sulfur nanoparticle synthesis mimicking environmental conditions of submarine shallow-water hydrothermal vents

Arsenic and sulfur mineralization is a natural phenomenon occurring in hydrothermal systems where parameters like temperature and organic matter(OM) can influence the mobilization of the toxic metalloid in marine environments. In the present study we analyze the influence of temperature and OM(particularly sulfur-containing additives) on As and S precipitation based on the recent discovery of As-rich nanoparticles in the hydrothermal system near the coast of the Greek island Milos. To this end, ...     

Difference in calcium ion precipitation between free and immobilized Halovibrio mesolongii HMY2

Biomineralization has become a research focus in wastewater treatment due to its much lower costs compared to traditional methods. However, the low sodium chloride (NaCl)-tolerance of bacteria limits applications to only water with low NaCl concentrations. Here, calcium ions in hypersaline wastewater (10% NaCl) were precipitated by free and immobilized Halovibrio mesolongii HMY2 bacteria and the differences between them were determined. The results show that calcium ions can be transformed into several types of calcium carbonate with a range of morphologies, abundant organic functional groups (C-H, C-O-C, C=O, etc), protein secondary structures (β-sheet, α-helix, 3₁₀ helix, and β-turn), P=O and S-H indicated by P2p and S2p, and more negative δ¹³C_PDB (‰) values (-16.8‰ to -18.4‰). The optimal conditions for the immobilized bacteria were determined by doing experiments with six factors and five levels and using response surface method. Under the action of two groups of immobilized bacteria prepared under the optimal conditions, by the 10^th day, Ca²⁺ ion precipitation ratios had increased to 79%-89% and 80%-88% with changes in magnesium ion cencentrations. Magnesium ions can significantly inhibit the calcium ion precipitation, and this inhibitory effect can be decreased under the action of immobilized bacteria. Minerals induced by immobilized bacteria always aggregated together, had higher contents of Mg, P, and S, lower stable carbon isotope values and less well-developed protein secondary structures. This study demonstrates an economic and eco-friendly method for recycling calcium ions in hypersaline wastewater, providing an easy step in the process of desalination.     

Bio-remediation of acephate–Pb(II) compound contaminants by Bacillus subtilis FZUL-33

Removal of Pb~(2+)and biodegradation of organophosphorus have been both widely investigated respectively. However, bio-remediation of both Pb~(2+)and organophosphorus still remains largely unexplored. Bacillus subtilis FZUL-33, which was isolated from the sediment of a lake, possesses the capability for both biomineralization of Pb~(2+)and biodegradation of acephate. In the present study, both Pb~(2+)and acephate were simultaneously removed via biodegradation and biomineralization in aqueous solutions.Batch experiments were conducted to study the influence of p H, interaction time and Pb~(2+)concentration on the process of removal of Pb2+. At the temperature of 25°C, the maximum removal of Pb~(2+)by B. subtilis FZUL-33 was 381.31 ± 11.46 mg/g under the conditions of p H 5.5, initial Pb~(2+)concentration of 1300 mg/L, and contact time of 10 min. Batch experiments were conducted to study the influence of acephate on removal of Pb~(2+)and the influence of Pb2+on biodegradation of acephate by B. subtilis FZUL-33. In the mixed system of acephate–Pb2+, the results show that biodegradation of acephate by B. subtilis FZUL-33 released PO43+, which promotes mineralization of Pb2+. The process of biodegradation of acephate was affected slightly when the concentration of Pb2+was below 100 mg/L. Based on the results, it can be inferred that the B. subtilis FZUL-33 plays a significant role in bio-remediation of organophosphorus-heavy metal compound contamination.     

Characterization and role of carbonic anhydrase in the calcification process of the azooxanthellate coral <Emphasis Type="Italic">Tubastrea aurea</Emphasis>

In zooxanthellate corals, the photosynthetic fixation of carbon dioxide and the precipitation of CaCO₃ are intimately linked both spatially and temporally making it difficult to study carbon transport mechanisms involved in each pathway. When studying Tubastrea aurea, a coral devoid of zooxanthellae, we can focus on carbon transport mechanisms involved only in the calcification process. We performed this study to characterize T. aurea carbonic anhydrase and to determine its role in the calcification process. We have shown that inhibition of tissular carbonic anhydrase activity affects the calcification rate. We have measured the activity of this enzyme both in the tissues and in the organix matrix extracted from the skeleton. Our results indicate that organic matrix proteins, which are synthesized by the calcifying tissues, are not only structural proteins, but they also play a crucial catalytic role by eliminating the kinetic barrier to interconversion of inorganic carbon at the calcification site. By immunochemistry we have demonstrated the presence of a protein both in the tissues and in the organic matrix, which shares common features with prokaryotic carbonic anhydrases.     

Biomineralogy of human urinary calculi (kidney stones) from some geographic regions of Sri Lanka

Kidney stones (urinary calculi) have become a global scourge since it has been recognized as one of the most painful medical problems. Primary causative factors for the formation of these stones are not clearly understood, though they are suspected to have a direct relationship to the composition of urine, which is mainly governed by diet and drinking water. Sixty nine urinary calculi samples which were collected from stone removal surgeries were analyzed chemically for their Na, K, Ca, Mg, Cu, Zn, Pb, Fe and phosphate contents. Structural and mineralogical properties of stones were studied by XRD and FT-IR methods. The mean contents of trace elements were 1348 mg kg⁻¹ (Na); 294 mg kg⁻¹ (K); 32% (Ca); 1426 mg kg⁻¹ (Mg); 8.39 mg kg⁻¹ (Mn); 258 mg kg⁻¹ (Fe); 67 mg kg⁻¹ (Cu); 675 mg kg⁻¹ (Zn); 69 mg kg⁻¹ (Pb); and 1.93% (PO₄³⁻). The major crystalline constituent in the calculi of Sri Lanka is calcium oxalate monohydrate. Principal component analysis was used to identify the multi element relationships in kidney stones. Three components were extracted and the first component represents positively correlated Na-K-Mg-PO₄³⁻ whereas the␣second components represent the larger positively weighted Fe–Cu–Pb. Ca–Zn correlated positively in the third component in which Mn–Cu correlated negatively. This study indicates that during the crystallization of human urinary stones, Ca shows more affinity towards oxalates whereas other alkali and alkaline earths precipitate with phosphates.Contribution from the Environmental Geology Research Group (EGRG), Department of Geology, University of Peradeniya, Sri Lanka.     

Bioremediation of strontium (Sr) contaminated aquifer quartz sand based on carbonate precipitation induced by Sr resistant Halomonas sp

Contamination of aquifers or sediments by radioactive strontium (⁹⁰Sr) is a significant environmental problem. In the present study, microbially induced calcite precipitation (MICP) was evaluated for its potential to remediate strontium from aquifer quartz sand. A Sr resistant urease producing Halomonas sp. was characterized for its potential role in bioremediation. The bacterial strain removed 80% of Sr from soluble-exchangeable fraction of aquifer quartz sand. X-ray diffraction detected calcite, vaterite and aragonite along with calcite-strontianite (SrCO₃) solid solution in bioremediated sample with indications that Sr was incorporated into the calcite. Scanning electron micrography coupled with energy-dispersive X-ray further confirmed MICP process in remediation. The study showed that MICP sequesters soluble strontium as biominerals and could play an important role in strontium bioremediation from both ecological and greener point of view.     

Enzymatic production of biosilica glass using enzymes from sponges: basic aspects and application in nanobiotechnology (material sciences and medicine)

Biomineralization, biosilicification in particular (i.e. the formation of biogenic silica, SiO₂), has become an exciting source of inspiration for the development of novel bionic approaches following “nature as model”. Siliceous sponges are unique among silica forming organisms in their ability to catalyze silica formation using a specific enzyme termed silicatein. In this study, we review the present state of knowledge on silicatein-mediated “biosilica” formation in marine sponges, the involvement of further molecules in silica metabolism and their potential application in nanobiotechnology and medicine. Werner E. G. Müller dedicated this study to Prof. Vera Gamulin (Rudjer Boskovic Institute, Zagreb, Croatia) in honour of her unique contributions in molecular evolution.     

Biomineralization-based conversion of carbon dioxide to calcium carbonate using recombinant carbonic anhydrase

Recently, as a mimic of the natural biomineralization process, the use of carbonic anhydrase (CA), which is an enzyme catalyzing fast reversible hydration of carbon dioxide to bicarbonate, has been suggested for biological conversion of CO₂ to valuable chemicals. While purified bovine CA (BCA) has been used in previous studies, its practical utilization in CO₂ conversion has been limited due to the expense of BCA preparation. In the present work, we investigated conversion of CO₂ into calcium carbonate as a target carbonate mineral by using a more economical, recombinant CA. To our knowledge, this is the first report of the usage of recombinant CA for biological CO₂ conversion. Recombinant α-type CA originating in Neisseria gonorrhoeae (NCA) was highly expressed as a soluble form in Escherichia coli. We found that purified recombinant NCA which showed comparable CO₂ hydration activity to commercial BCA significantly promoted formation of solid CaCO₃ through the acceleration of CO₂ hydration rate, which is naturally slow. In addition, the rate of calcite crystal formation was also accelerated using recombinant NCA. Moreover, non-purified crude recombinant NCA also showed relatively significant ability. Therefore, recombinant CA could be an effective, economical biocatalyst in practical CO₂ conversion system.     

Alteration of shell nacre micromorphology in blue mussel Mytilus edulis after exposure to free-ionic silver and silver nanoparticles

This study describes the morphology of inner shell surface (ISS) of the blue mussel Mytilus edulis Linnaeus after short-term exposures to radiolabeled silver in free-ionic (^110mAg⁺) and engineered nanoparticulate (^110mAgNPs, <40 nm) phases. Radiolabeled silver in starting solutions was used in a similar low concentration (∼15 Bq mL⁻¹) for both treatments. After exposure experiments radiolabeled silver was leached from the ISS using HCl. It concentration for shells from both treatments was ∼0.5 Bq mL⁻¹. Whole ISS of young individuals and prismatic layer of adults showed no evidence of any major alteration process after silver uptake. However, the nacre portion of adult mussels exposed to both treatments revealed distinct doughnut shape structures (DSS) formed by calcium carbonate micrograins that covered the surface of aragonite tablets. Scanning electron microscope (SEM) imaging revealed the existence of only minor differences in DSS morphology between mussels exposed to Ag⁺ and AgNPs. From literature survey, DSS were also found in bivalves exposed to Cd²⁺. The DSS occurring in a specimen of a field-collected bivalve is also shown. Formation of distinctive DSS can be explained by a disturbance of the shell calcification mechanism. Although the occurrence of DSS is not exclusively associated with metal bioavailability to the mussels, the morphology of DSS seems to be linked to the speciation of the metal used in the uptake experiments.