Jin-lan Xia

The effect of mixotrophy on microalgal growth, lipid content, and expression levels of three pathway genes in Chlorella sorokiniana

Nannochloropsis oculata CCMP 525, Dunaliella salina FACHB 435, and Chlorella sorokiniana CCTCC M209220 were compared in mixotrophic and photoautotrophic cultures in terms of growth rate, protein, and lipid content. Growth improved in glucose, and the biomass productivities of N. oculata, D. salina, and C. sorokiniana were found to be 1.4-, 2.2- and 4.2-fold that observed photoautotrophically. However, biomass and lipid production decreased at the highest glucose concentrations. Meanwhile, the content of protein and lipid were significantly augmented for mixotrophic conditions at least for some species. C. sorokiniana was found to be well suited for lipid production based on its high biomass production rate and lipid content reaching 51% during mixotrophy. Expression levels of accD (heteromeric acetyl-CoA carboxylase beta subunit), acc1 (homomeric acetyl-CoA carboxylase), rbcL (ribulose 1, 5-bisphosphate carboxylase/oxygenase large subunit) genes in C. sorokiniana were studied by real-time PCR. Increased expression levels of accD reflect the increased lipid content in stationary phase of mixotrophic growth, but expression of the acc1 gene remains low, suggesting that this gene may not be critical to lipid accumulation. Additionally, reduction of expression of the rbcL gene during mixotrophy indicated that utilization of glucose was found to reduce the role of this gene and photosynthesis.

Sulfur oxidation activities of pure and mixed thermophiles and sulfur speciation in bioleaching of chalcopyrite

The sulfur oxidation activities of four pure thermophilic archaea Acidianus brierleyi (JCM 8954), Metallosphaera sedula (YN 23), Acidianus manzaensis (YN 25) and Sulfolobus metallicus (YN 24) and their mixture in bioleaching chalcopyrite were compared. Meanwhile, the relevant surface sulfur speciation of chalcopyrite leached with the mixed thermophilic archaea was investigated. The results showed that the mixed culture, with contributing significantly to the raising of leaching rate and accelerating the formation of leaching products, may have a higher sulfur oxidation activity than the pure cultures, and jarosite was the main passivation component hindering the dissolution of chalcopyrite, while elemental sulfur seemed to have no influence on the dissolution of chalcopyrite. In addition, the present results supported the former speculation, i.e., covellite might be converted from chalcocite during the leaching experiments, and the elemental sulfur may partially be the derivation of covellite and chalcocite.

Physiological evaluation of a new Chlorella sorokiniana isolate for its biomass production and lipid accumulation in photoautotrophic and heterotrophic cultures.

A novel green unicellular microalgal isolate from the freshwater of the Inner Mongolia Province of China and named as CCTCC M209220, grows between pH 6 and 11 and temperatures of 20–35°C with optimal conditions at pH 9 and 30°C. Morphological features and the phylogenetic analysis for the 18S rRNA gene reveal that the isolate is a Chlorella sorokiniana strain. A nitrogen source test reveals that this strain can grow well with nitrate and urea, but not ammonium. The strain can grow heterotrophically with glucose as the carbon source and accumulates lipid content as high as 56% (w/w) dry weight after 7 days in high glucose concentrations compared to 19% lipids achieved in 30 days of photoautotrophic culture. The relative neutral lipid content as a fraction of the total lipid is also much higher in heterotrophic culture as compared to photoautotrophic culture. Biotechnol. Bioeng. 2012; 109:1958–1964.

Adhesion forces between cells of Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans or Leptospirillum ferrooxidans and chalcopyrite.

The efficiency of copper leaching is improved by bacteria attached to chalcopyrite. Therefore, the study of the attachment mechanism to control leaching is important. The adhesion of three species of leaching microorganisms including Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans and Leptospirillum ferrooxidans to chalcopyrite was investigated by using atomic force microscopy (AFM). The forces were measured with tip-immobilized cells approached to and retracted from the mineral. The results show that both the surface charge and the hydrophobicity of bacteria cells influence the adhesion force. Furthermore, the adhesion force decreased in case the extracellular polymeric substances (EPS) had been removed. In addition, the data indicate that the amount of attached cells increased with increasing adhesion force.

A new strain Acidithiobacillus albertensis BY-05 for bioleaching of metal sulfides ores

Abstract An acidophilic, rod-shaped Gram-negative sulfur oxidizing strain BY-05 was isolated from an acid mine drainage of copper ore in Baiyin area, Gansu Province, China. Ultrastructural studies show that the isolate has a tuft of polar flagella and possesses sulfur granules with clear membrane adhering to the cell innermembrane. Physiological study shows that this isolate grows autotrophically and aerobically by oxidizing S0 and reduced inorganic sulfur compounds (S2O2-3, S2O2-4, S2- and ZnS) with the optimum growth at pH 3.5–4.0 and at the temperature range of 25–30 °C. The 16S rRNA gene sequence (DQ 423683) of strain BY-05 has 100% sequence similarity to that of Acidithiobacillus albertensis (DSM 14366). So it is identified and named as A. albertensis BY-05. Bioleaching experiments with this new strain show that it can play an important role in recovery of metals from chalcopyrite and sphalerite.

An improved colony PCR procedure for genetic screening of Chlorella and related microalgae

A colony PCR technique was applied for both genomic and chloroplast DNA in the green microalgae Chlorella. Of five different lysis buffers, Chelex-100 was superior for DNA extraction, PCR and DNA storage. It also was insensitive to variations in cell density. The conditions established for an improved PCR formulation are applicable for screening of genetically-engineered transformants as well as bioprospecting of natural microalgal isolates. Besides multiple Chlorella species, we also demonstrate the efficacy of Chelex-100 for colony PCR with a number of other microalgal strains, including Chlamydomonas reinhardtii, Dunaliella salina, Nannochloropsis sp., Coccomyxa sp., and Thalassiosira pseudonana.

Comparative study on biosorption of Pb(II) and Cr(VI) by Synechococcus sp.

Abstract The comparative study on adsorptions of Pb(II) and Cr(VI) ions by free cells and immobilized cells of Synechococcus sp. was performed, in which different aspects including Zeta potential of the cells, the influence of pH, temperature and initial concentration of metal ions, as well as adsorption kinetics and mechanism were referred. The lyophilized free cells have a surface isoelectric point at pH 3, and the correlative experiment indicates that there is an electrostatic adsorption feature of Cr(VI) and Pb(II). The immobilization of the free cells by Ca-alginate does not significantly modify the adsorption features of the biosorbent. The absorption processes of Cr(VI) and Pb(II) on both free and immobilized cells are apparently affected by pH and the initial concentration of metal ions in the bulk solution, but are much weakly affected by temperature in the test range of 10−50°C. The slow course of biosorption follows the first order kinetic model, the adsorption of Pb(II) obeys both Langmuir and Freundlich isotherm models, while the adsorption of Cr(VI) obeys only Freundlich model. FT-IR results indicate that carboxylic, alcoholic, amide and amino groups are responsible for the binding of the metal ions, and reduction of Cr(VI) to Cr(III) takes place after Cr(VI) adsorbs electrostatically onto the surface of the biosorbents.

Comparative study on effects of Tween-80 and sodium isobutyl-xanthate on growth and sulfur-oxidizing activities of Acidithiobacillus albertensis BY-05

Abstract Effects of two typical surfactants, Tween-80 and sodium isobutyl-xanthate (NaIBX), with different concentrations on the growth and sulfur-oxidizing activities of a new strain Acidithiobacillus albertensis BY-05, an acidophilic sulfur-oxidizing bacterium, were investigated. The results indicate that both surfactants can enhance the growth and sulfur-oxidizing activities of A. albertensis BY-05 only at some special concentrations, e.g., 10−4–10−8 g/L for NaIBX and lower than 10−8 g/L for Tween-80, but were inhibited and even harmful at higher concentrations. Both surfactants can not be metabolized by A. albertensis BY-05. The contact between the bacteria and the sulfur particles may be dependent upon both the extracellular substance and the surfactants, both of which provide the amphiphilic environment improving the attachment for bacteria to the sulfur particles surface. These data could be significant for enlarging the applications of both A. albertensis BY-05 and some typical surfactants for industrial bioleaching of sulfides minerals.

Growth and surface properties of new thermoacidophilic Archaea strain Acidianus manzaensis YN-25 grown on different substrates

Abstract The growth and surface properties of new thermoacidophilic Archaea strain Acidianus manzaensis YN-25 isolated from an acid hot spring in Tengchong, Yunnan Province, China were investigated cultured on different substrates including soluble substrate ferrous sulfate and nonsoluble solid substrates S 0 , pyrite and chalcopyrite. The growth characteristics of the cells in each substrate were characterized with the changes in cell number, pH, E h , and concentrations of Fe 2+ or SO 4 2− , or ratios of [Fe 2+ ] to [Fe 3+ ], and the surface properties were characterized and analyzed in terms of Zeta-potential, hydrophobicity, and surface FT-IR spectra of the cells. The results show that the cells grown on solid substrates have higher value of isoelectric points. They are more hydrophobic and express more surface proteins than ferrous sulfate grown cells.

Relationships among bioleaching performance, additional elemental sulfur, microbial population dynamics and its energy metabolism in bioleaching of chalcopyrite

Abstract To estimate the relationships among bioleaching performance, additional elemental sulfur (S0), microbial population dynamics and its energy metabolism, bioleaching of chalcopyrite by three typical sulfur- and/or iron-oxidizing bacteria, Acidithiobacillus ferrooxidans, Leptospirillum ferriphilum and Acidithiobacillus thiooxidans with different levels of sulfur were studied in batch shake flask cultures incubated at 30°C. Copper dissolution capability (71%) was increased with the addition of 3.193 g/L S0, compared to that (67%) without S0. However, lower copper extraction was obtained in bioleaching with excessive sulfur. Microbial population dynamics during chalcopyrite bioleaching process was monitored by using PCR-restriction fragment length polymorphism (PCR-RFLP). Additional S0 accelerated the growth of sulfur-oxidizing bacteria, inhibited the iron-oxidizing metabolism and led to the decrease of iron-oxidizing microorganisms, finally affected iron concentration, redox potential and bioleaching performance. It is suggested that mixed iron and sulfur-oxidizing microorganisms with further optimized additional S0 concentration could improve copper recovery from chalcopyrite.