Cadmium-resistant Streptomyces stimulates phytoextraction potential of Crotalaria juncea L. in cadmium-polluted soil.

This work evaluated the competence of two strains of cadmium (Cd)-resistant Streptomyces, namely Streptomyces rapamycinicus K5PN1, an indole-3-acetic acid (IAA) producer, and Streptomyces cyaneus 11-10SHTh, a siderophore producer on promoting Cd phytoextraction by sunn hemp. The results showed that S. rapamycinicus improved root elongation of sunn hemp seedlings under Cd stress conditions. S. rapamycinicus and S. cyaneus were colonized on the root surface of sunn hemp at concentrations of 2.3 × 104 and 6.4 × 103 CFU g-1 root fresh weight, respectively. The results of pot-culture experiments showed that S. rapamycinicus increased the root and shoot lengths, and dry biomass of sunn hemp planted in high Cd-contaminated soil. The Cd concentration in the leaves of sunn hemp inoculated with S. cyaneus (73.82 ± 2.20 mg kg-1 plant dry wt) was higher than that of plants with S. rapamycinicus inoculation and the uninoculated control. The phytoextraction of Cd by sunn hemp was significantly increased with Cd-resistant Streptomyces inoculation. In conclusion, both strains of Cd-resistant Streptomyces had potential on enhancing Cd phytoextraction efficiency of sunn hemp. Our study suggests the application of Cd-resistant Streptomyces can improve Cd phytoextraction by sunn hemp for restoration of Cd-polluted sites.

Our study demonstrated the potential of two strains of Cd-resistant Streptomyces to stimulate Cd phytoextraction from soil by sunn hemp. Cadmium-resistant Streptomyces strongly stimulated Cd uptake and accumulation in sunn hemp planted in high level of Cd-polluted soil, supporting sunn hemp to be a superior Cd accumulator.

Effect of phosphorylated polyvinyl alcohol matrix size of cell entrapment on partial nitrification of ammonia in wastewater.

Partial nitrification is known as first and critical step for autotrophic nitrogen removal in high strength nitrogenous wastewater. Phosphorylated polyvinyl alcohol gel entrapment was used for suppressing oxygen to nitrite-oxidizing bacteria (NOB) in the gel matrix. The study investigated the effect of the size of gel matrix on partial nitrification. Results show that ammonia-oxidizing bacteria (AOB) proportion in the inoculum rather than the size of gel matrix governed ammonia oxidation. Nitrite oxidation depended on the size of gel matrix not the relative proportions of NOB and AOB in the inoculum. Larger size of gel matrix lead to less in situ oxygen penetration and available for NOB resulting in higher nitrite accumulation. This finding gains a better understanding of using suitable inoculum to control partial nitrification that is beneficial for the preparation of anaerobic ammonium oxidation-suited effluent.

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.

Bio-based dispersants for fuel oil spill remediation based on the Hydrophilic-Lipophilic Deviation (HLD) concept and Box-Behnken design.

The high density and viscosity of fuel oil leads to its prolonged persistence in the environment and causes widespread contamination. Dispersants with a low environmental impact are necessary for fuel oil spill remediation. This study aimed to formulate bio-based dispersants by mixing anionic biosurfactant (lipopeptides from Bacillus subtilis GY19) with nonionic oleochemical surfactant (Dehydol LS7TH). The synergistic effect of the anionic-nonionic surfactant mixture produced a Winsor Type III microemulsion, which promoted petroleum mobilization. The hydrophilic-lipophilic deviation (HLD) equations for ionic and nonionic surfactant mixtures were compared, and it was found that the ionic equation was applicable for the calculation of lipopeptides and Dehydol LS7TH concentrations. The best formula contained 6.6% w/v lipopeptides and 11.9% w/v Dehydol LS7TH in seawater, and its dispersion effectiveness for bunker fuels A and C was 92% and 78%, respectively. The application of bio-based dispersants in water sources was optimized by Box-Behnken design. The efficiency of the bio-based dispersant was affected by the dispersant-to-oil ratios (DORs) but not by the water salinity. A suitable range of DORs for different oil contamination levels could be identified from the response surface plot. The dispersed fuel oil was further degraded by adding an oil-degrading bacterial consortium to the chemically enhanced water accommodated fractions (CEWAFs). After 7 days of incubation, the concentration of fuel oil was reduced from 3692 mg/L to 356 mg/L (88% removal efficiency). On the other hand, the abiotic control removed less than 40% fuel oil from the CEWAFs. This bio-based dispersant had an efficiency comparable to that of a commercial dispersant. The process of dispersant formulation and optimization could be applied to other surfactant mixtures.

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.