Copper and chromium removal from industrial sludge by a biosurfactant-based washing agent and subsequent recovery by iron oxide nanoparticles.

Industrial wastewater treatment generates sludge with high concentrations of metals and coagulants, which can cause environmental problems. This study developed a sequential sludge washing and metal recovery process for industrial sludge containing > 4500 mg/kg Cu and > 5000 mg/kg Cr. The washing agent was formulated by mixing glycolipid, lipopeptide, and phospholipid biosurfactants from Weissella cibaria PN3 and Brevibacterium casei NK8 with a chelating agent, ethylenediaminetetraacetic acid (EDTA). These biosurfactants contained various functional groups for capturing metals. The optimized formulation by the central composite design had low surface tension and contained relatively small micelles. Comparable Cu and Cr removal efficiencies of 37.8% and 38.4%, respectively, were obtained after washing the sludge by shaking with a sonication process at a 1:4 solid-to-liquid ratio. The zeta potential analysis indicated the bonding of metal ions on the surface of biosurfactant micelles. When 100 g/L iron oxide nanoparticles were applied to the washing agent without pH adjustment, 83% Cu and 100% Cr were recovered. In addition, X-ray diffraction and X-ray absorption spectroscopy of the nanoparticles showed the oxidation of nanoparticles, the reduction of Cr(V) to the less toxic Cr(III), and the absorption of Cu. The recovered metals could be further recycled, which will be beneficial for the circular economy.

Formulation of a glycolipid:lipopeptide mixture as biosurfactant-based dispersant and development of a low-cost glycolipid production process.

Biosurfactant-based dispersants were formulated by mixing glycolipids from Weissella cibaria PN3 and lipopeptides from Bacillus subtilis GY19 to enhance the synergistic effect and thereby achieve hydrophilic-lipophilic balance. The proportions of each biosurfactant and dispersant-to-oil ratios (DORs) were varied to obtain the appropriated formulations. The most efficient glycolipid:lipopeptide mixtures (F1 and F2) had oil displacement activities of 81-88% for fuel and crude oils. The baffled flask test of these formulations showed 77-79% dispersion effectiveness at a DOR of 1:25. To reduce the cost of the dispersant, this study optimized the glycolipid production process by using immobilized cells in a stirred tank fermenter. Semicontinuous glycolipid production was carried out conveniently for 3 cycles. Moreover, food wastes, including waste coconut water and waste frying oil, were found to promote glycolipid production. Glycolipids from the optimized process and substrates had similar characteristics but 20-50% lower cost than those produced from basal medium with soybean oil in shaking flasks. The lowest cost dispersant formulation (F2*) contained 10 g/L waste-derived cell-bound glycolipid and 10 g/L lipopeptide and showed high dispersion efficiency with various oils. Therefore, this biosurfactant-based dispersant could be produced on a larger scale for further application.

Physiological changes in Rhodococcus ruber S103 immobilized on biobooms using low-cost media enhance stress tolerance and crude oil-degrading activity.

For economic feasibility, sugarcane molasses (0.5%, w/v) containing K2HPO4 (0.26%, w/v) and mature coconut water, low value byproducts, were used in cultivation of Rhodococcus ruber S103 for inoculum production and immobilization, respectively. Physiological changes of S103 grown in low-cost media, including cell hydrophobicity, saturated/unsaturated ratio of cellular fatty acids and biofilm formation activity, enhanced stress tolerance and crude oil biodegradation in freshwater and even under high salinity (5%, w/v). Biobooms comprised of S103 immobilized on polyurethane foam (PUF) was achieved with high biomass content (1010 colony-forming units g-1 PUF) via a scale-up process in a 5-L modified fluidized-bed bioreactor within 3 days. In a 500-L mesocosm, natural freshwater was spiked with crude oil (72 g or 667 mg g-1 dry biobooms), and a simulated wave was applied. Biobooms could remove 100% of crude oil within only 3 days and simultaneously biodegraded 60% of the adsorbed oil after 7 days when compared to boom control with indigenous bacteria. In addition, biobooms had a long shelf-life (at least 100 days) with high biodegradation activity (85.2 ± 2.3%) after storage in 10% (w/v) skimmed milk at room temperature. This study demonstrates that the low-cost production of biobooms has potential for future commercial bioremediation.

Formulation of Bio-Based Washing Agent and Its Application for Removal of Petroleum Hydrocarbons From Drill Cuttings Before Bioremediation.

Drill cuttings from petroleum exploration and production sites can cause diverse environmental problems. Total petroleum hydrocarbons (TPHs) are a major pollutant from the use of polyolefin-based mud. As an alternative to incineration, this study investigated the application of surfactant-enhanced washing technology prior to bioremediation. The washing step was necessary because the initial TPH concentrations were quite high at approximately 15% (w/w). Washing agents were formulated by varying the concentration of lipopeptide biosurfactant (in foamate or cell-free broth), Dehydol LS7TH (fatty alcohol ethoxylate 7EO, oleochemical surfactant) and butanol (as a lipophilic linker) at different salinities. The most efficient formula produced a Winsor Type I microemulsion (oil-in-water microemulsion) with polyolefin and contained only 20% (v/v) foamate and 2% (v/v) Dehydol LS7TH in water. Due to the synergistic behavior between the anionic lipopeptides and non-ionic Dehydol LS7TH, the formula efficiently removed 92% of the TPHs from the drill cuttings when applied in a jar test. To reduce the cost, the concentrations of each surfactant should be reduced; thus, the formula was optimized by the simplex lattice mixture design. In addition, cell-free broth, at a pH of 10, containing 3.0 g/L lipopeptides was applied instead of foamate because it was easy to prepare. The optimized formula removed 81.2% of the TPHs and contained 72.0% cell-free broth and 1.4% Dehydol LS7TH in water. A 20-kg soil washing system was later tested where the petroleum removal efficiency decreased to 70.7% due to polyolefin redeposition during separation of the washing solution. The remaining TPHs (4.5%) in the washed drilled cuttings were further degraded by a mixture of Marinobacter salsuginis RK5, Microbacterium saccharophilum RK15 and Gordonia amicalis JC11. To promote TPH biodegradation, biochar and fertilizer were applied along with bacterial consortia in a microcosm experiment. After 49-day incubation, the TPHs were reduced to 0.9% by both physical and biological mechanisms, while the TPHs in the unamended samples remained unaffected. With the use of the formulated bio-based washing agent and bioremediation approach, the on-site treatment of drill cuttings could be conducted with an acceptable cost and low environmental impacts.

Reuse of Immobilized Weissella cibaria PN3 for Long-Term Production of Both Extracellular and Cell-Bound Glycolipid Biosurfactants.

Lactic acid bacteria (LABs) are generally recognized as safe (GRAS), and therefore, LAB biosurfactants are beneficial with negligible negative impacts. This study aims to maintain the biosurfactant producing activity of an LAB strain, Weissella cibaria PN3, by immobilizing the bacterial cells on a commercial porous carrier. For biosurfactant production, 2% soybean oil was used as the carbon source. After 72 h, immobilized cells were reused by replacing production medium. The extracellular and cell-bound biosurfactants were extracted from the resulting cell-free broth and cell pellets, respectively. SEM images of used immobilizing carriers showed increased surface roughness and clogged pores over time. Thus, the immobilizing carriers were washed in PBS buffer (pH 8.0) before reuse. To maintain biosurfactant production activity, immobilized cells were reactivated every three production cycles by incubating the washed immobilizing carriers in LB medium for 48 h. The maximum yields of purified extracellular (1.46 g/L) and cell-bound biosurfactants (1.99 g/L) were achieved in the 4th production cycle. The repeated biosurfactant production of nine cycles were completed within 1 month, while only 2 g of immobilized cells/L were applied. The extracellular and cell-bound biosurfactants had comparable surface tensions (31 - 33 mN/m); however, their CMC values were different (1.6 and 3.2 g/L, respectively). Both biosurfactants had moderate oil displacement efficiency with crude oil samples but formed emulsions well with gasoline, diesel, and lavender, lemongrass and coconut oils. The results suggested that the biosurfactants were relatively hydrophilic. In addition, the mixing of both biosurfactants showed a synergistic effect, as seen from the increased emulsifying activity with palm, soybean and crude oils. The biosurfactants at 10 - 16 mg/mL showed antimicrobial activity toward some bacteria and yeast but not filamentous fungi. The molecular structures of these biosurfactants were characterized by FTIR as different glycolipid congeners. The biosurfactant production process by immobilized Weissella cibaria PN3 cells was relatively cheap given that two types of biosurfactants were simultaneously produced and no new inoculum was required. The acquired glycolipid biosurfactants have high potential to be used separately or as mixed biosurfactants in various products, such as cleaning agents, food-grade emulsifiers and cosmetic products.

Formulation of crude oil spill dispersants based on the HLD concept and using a lipopeptide biosurfactant.

Solvent-free dispersants for crude oil spills were formulated based on the hydrophilic-lipophilic deviation (HLD) concept and using lipopeptides from Bacillus sp. GY19. The lipopeptides were recovered and concentrated from cell-free broth by foam fractionation and freeze-drying. They had good surface activity under varying temperatures, pH and NaCl levels. Moreover, the lipopeptides had low toxicity to copepods (LC50 1174mg/L) and whiteleg shrimp (LC50 1050mg/L). The characteristic curvature (Cc) of the lipopeptides showed that they were more hydrophobic (Cc 4.93) than sodium dihexyl sulfosuccinate (SDHS, Cc -0.92). The HLD equation was used to calculate the lipopeptide and the SDHS fractions in the dispersant formulations according to the equivalent alkane carbon number (EACN) of hydrocarbons and seawater salinity. The molar fraction of lipopeptides increased with increasing EACN. The lipopeptide-SDHS mixtures formed microemulsion Type III with specific hydrocarbons and crude oils. Oil displacement and baffled flask tests showed that the formulations reduced the interfacial tension and solubilized crude oil in the water column at higher efficiency than commercial dispersants or lipopeptides alone. In summary, the effectiveness of the lipopeptide-based dispersant corresponded to the optimal HLD.

Airlift bioreactor containing chitosan-immobilized Sphingobium sp. P2 for treatment of lubricants in wastewater.

An internal loop airlift bioreactor containing chitosan-immobilized Sphingobium sp. P2 was applied for the removal of automotive lubricants from emulsified wastewater. The chitosan-immobilized bacteria had higher lubricant removal efficiency than free and killed-immobilized cells because they were able to sorp and degrade the lubricants simultaneously. In a semi-continuous batch experiment, the immobilized bacteria were able to remove 80-90% of the 200 mg L(-1) total petroleum hydrocarbons (TPH) from both synthetic and carwash wastewater. The internal loop airlift bioreactor, containing 4 g L(-1) immobilized bacteria, was later designed and operated at 2.0 h HRT (hydraulic retention time) for over 70 days. At a steady state, the reactor continuously removed 85±5% TPH and 73±11% chemical oxygen demand (COD) from the carwash wastewater with 25-200 mg L(-1) amended lubricant. The internal loop airlift reactor's simple operation and high stability demonstrate its high potential for use in treating lubricants in emulsified wastewater from carwashes and other industries.