Simultaneous production of algal biomass and lipid by heterotrophic cultivation of linoleic acid-rich oleaginous microalga Chlorella sorokiniana using high acetate dosage.

The low-cost carbon source, acetate, was utilized to feed a linoleic acid-rich Chlorella sorokiniana for microalgal biomass and lipid accumulation. Remarkably high tolerance capability to high acetate dosage up to 30 g/L was observed, with heterotrophy being the preferred trophic mode for algal growth and lipogenesis when supplemented 20 g/L acetate. Transcriptome analysis revealed a marked activation of pathways involved in acetate bioconversion and lipogenesis upon exposure to high-level of acetate. However, the enhancement of photorespiration inhibited photosynthesis, which ultimately led to a decrease in biomass and lipid under mixotrophy. Heterotrophic acetate-feeding generated more superior amino acid profiling of algal biomass and a predominant linoleic acid content (50 %). Heterotrophic repeat fed-batch strategy in 5 L fermenter significantly increased the growth performance and lipid titer, with the highest levels achieved being 23.4 g/L and 7.0 g/L, respectively. This work provides a viable approach for bio-products production through acetate-based heterotrophic algal cultivation.

D-Lactic acid production from agricultural residues by membrane integrated continuous fermentation coupled with B vitamin supplementation.

BACKGROUND: D-Lactic acid played an important role in the establishment of PLA as a substitute for petrochemical plastics. But, so far, the D-lactic acid production was limited in only pilot scale, which was definitely unable to meet the fast growing market demand. To achieve industrial scale D-lactic acid production, the cost-associated problems such as high-cost feedstock, expensive nutrient sources and fermentation technology need to be resolved to establish an economical fermentation process. RESULTS: In the present study, the combined effect of B vitamin supplementation and membrane integrated continuous fermentation on D-lactic acid production from agricultural lignocellulosic biomass by Lactobacillus delbrueckii was investigated. The results indicated the specific addition of vitamins B1, B2, B3 and B5 (VB1, VB2, VB3 and VB5) could reduce the yeast extract (YE) addition from 10 to 3 g/l without obvious influence on fermentation efficiency. By employing cell recycling system in 350 h continuous fermentation with B vitamin supplementation, YE addition was further reduced to 0.5 g/l, which resulted in nutrient source cost reduction of 86%. A maximum D-lactate productivity of 18.56 g/l/h and optical purity of 99.5% were achieved and higher than most recent reports. CONCLUSION: These findings suggested the novel fermentation strategy proposed could effectively reduce the production cost and improve fermentation efficiency, thus exhibiting great potential in promoting industrial scale D-lactic acid production from lignocellulosic biomass.

Quantum dots are conventionally applicable for wide-profiling of wall polymer distribution and destruction in diverse cells of rice.

Plant cell walls represent enormous biomass resources for biofuels, and it thus becomes important to establish a sensitive and wide-applicable approach to visualize wall polymer distribution and destruction during plant growth and biomass process. Despite quantum dots (QDs) have been applied to label biological specimens, little is reported about its application in plant cell walls. Here, semiconductor QDs (CdSe/ZnS) were employed to label the secondary antibody directed to the epitopes of pectin or xylan, and sorted out the optimal conditions for visualizing two polysaccharides distribution in cell walls of rice stem. Meanwhile, the established QDs approach could simultaneously highlight wall polysaccharides and lignin co-localization in different cell types. Notably, this work demonstrated that the QDs labeling was sensitive to profile distinctive wall polymer destruction between alkali and acid pretreatments with stem tissues of rice. Hence, this study has provided a powerful tool to characterize wall polymer functions in plant growth and development in vivo, as well as their distinct roles during biomass process in vitro.

Replacing process water and nitrogen sources with biogas slurry during cellulosic ethanol production.

BACKGROUND: Environmental issues, such as the fossil energy crisis, have resulted in increased public attention to use bioethanol as an alternative renewable energy. For ethanol production, water and nutrient consumption has become increasingly important factors being considered by the bioethanol industry as reducing the consumption of these resources would decrease the overall cost of ethanol production. Biogas slurry contains not only large amounts of wastewater, but also the nutrients required for microbial growth, e.g., nitrogen, ammonia, phosphate, and potassium. Therefore, biogas slurry is an attractive potential resource for bioethanol production that could serve as an alternative to process water and nitrogen sources. RESULTS: In this study, we propose a method that replaces the process water and nitrogen sources needed for cellulosic ethanol production by Zymomonas mobilis with biogas slurry. To test the efficacy of these methods, corn straw degradation following pretreatment with diluted NaOH and enzymatic hydrolysis in the absence of fresh water was evaluated. Then, ethanol fermentation using the ethanologenic bacterial strain Z. mobilis ZMT2 was conducted without supplementing with additional nitrogen sources. After pretreatment with 1.34% NaOH (w/v) diluted in 100% biogas slurry and continuous enzymatic hydrolysis for 144 h, 29.19 g/L glucose and 12.76 g/L xylose were generated from 30 g dry corn straw. The maximum ethanol concentration acquired was 13.75 g/L, which was a yield of 72.63% ethanol from the hydrolysate medium. Nearly 94.87% of the ammonia nitrogen was depleted and no nitrate nitrogen remained after ethanol fermentation. The use of biogas slurry as an alternative to process water and nitrogen sources may decrease the cost of cellulosic ethanol production by 10.0-20.0%. By combining pretreatment with NaOH diluted in biogas slurry, enzymatic hydrolysis, and ethanol fermentation, 56.3 kg of ethanol was produced by Z. mobilis ZMT-2 through fermentation of 1000 kg of dried corn straw. CONCLUSIONS: In this study, biogas slurry replaced process water and nitrogen sources during cellulosic ethanol production. The results suggest that biogas slurry is a potential alternative to water when pretreating corn straw and, thus, has important potential applications in cellulosic ethanol production from corn straw. This study not only provides a novel method for utilizing biogas slurry, but also demonstrates a means of reducing the overall cost of cellulosic ethanol.

A synergistic combination of nutrient reclamation from manure and resultant hydrochar upgradation by acid-supported hydrothermal carbonization.

Cattle manure was hydrothermally carbonized in acid solutions (0-2% HCl), then nutrient concentration in liquid product and physicochemical properties of hydrochar were characterized to investigate the effects of acid addition on hydrochar properties and nutrient recovery from manure. Results showed that hydrothermal carbonization (HTC) in 2% HCl extracted almost 100% and 63.38% of phosphorus and nitrogen, respectively; specifically, >90% of the extracted phosphorus was PO4-P in liquid from HTC with acid addition, and increasing amount of extracted nitrogen was NH4-N with increasing acid addition. Generally, higher heating value, surface area, total pore volume, fixed carbon, atomic ratios of H/C and O/C were increased in hydrochars from HTC with acid addition, while yield, volatile matter, contents of nitrogen, sulfur and oxygen of these hydrochars were decreased. These results indicated that HTC with acid addition could simultaneously facilitate nutrient recovery from manure and resulting hydrochar upgradation.

Calcium-rich biochar from the pyrolysis of crab shell for phosphorus removal.

Calcium-rich biochars (CRB) prepared through pyrolysis of crab shell at various temperatures were characterized for physicochemical properties and P removal potential. Elemental analysis showed that CRB was rich in calcium (22.91%-36.14%), while poor in carbon (25.21%-9.08%). FTIR, XRD and TG analyses showed that calcite-based CRB was prepared at temperature ≤600 °C, while lime-based CRB was prepared at temperature ≥700 °C. Phosphorus removal experiment showed that P removal efficiencies in 80 mg P/L phosphate solution and biogas effluent ranged from 26% to 11%, respectively, to about 100% and 63%, respectively, depending on the pyrolysis temperature of the resulting biochar. Specifically, compared to common used CaCO3 and Ca(OH)2, P removal potential of calcite-based CRB was much higher than that of CaCO3; while that of lime-based CRB was close to that of Ca(OH)2. These results suggested that CRB was competent for P removal/recovery from wastewater.

Terrisporobacter petrolearius sp. nov., isolated from an oilfield petroleum reservoir.

A Gram-staining-positive, spore-forming, strictly anaerobic bacterium, designated strain LAM0A37T, was isolated from enrichment samples collected from a petroleum reservoir in Shengli oilfield. Cells of strain LAM0A37T were rod-shaped and motile by peritrichous flagella. The optimal temperature and pH for growth were 40 °C and 7.0­7.5, respectively. The strain did not require NaCl for growth but tolerated up to 3 % (w/v) NaCl. Strain LAM0A37T was able to utilize glucose, fructose, maltose, xylose, sorbitol, cellobiose, melibiose and melezitose as sole carbon sources. Sulfite was used as an electron acceptor. The main products of glucose fermentation were acetate and CO2. The predominant fatty acid was C16 : 0 (23.6 %). The main polar lipid profile comprised of five glycolipids, six phospholipids and two lipids. No menaquinone was detected. The genomic DNA G+C content was 27.1 ± 0.2 mol% as determined by the T m method. Analysis of the 16S rRNA gene sequence indicated that the isolate was a member of the genus Terrisporobacter, and was most closely related to Terrisporobacter glycolicus JCM 1401T and Terrisporobacter mayombei DSM 6539T with 98.3 % 16S rRNA gene sequence similarity to both. DNA­DNA hybridization values between strain LAM0A37T and type strains of Terrisporobacter glycolicus and Terrisporobacter mayombei were 45.6 ± 0.3 % and 38.3 ± 0.4 %, respectively. Based on phenotypic, phylogenetic and chemotaxonomic characteristics, strain LAM0A37T is suggested to represent a novel species of the genus Terrisporobacter, for which the name Terrisporobacter petrolearius sp. nov. is proposed. The type strain is LAM0A37T ( = ACCC 00740T = JCM 19845T).

Immobilization of phosphorus in cow manure during hydrothermal carbonization.

The surplus of manure phosphorus (P) with increasing livestock production might pose a risk of P loss to the environment due to the high mobility of P in manure. Thus, there is an increasing need to mitigate P loss from manure. This study aimed to investigate the effect of hydrothermal carbonization (HTC) on the immobilization of P in cow manure. The results demonstrated that the P content in cow manure was increased substantially by ∼20% after HTC, while the water-extractable P (WEP) and Mehlich-3-extractable P (MEP) in manure was reduced significantly by >80% and 50%, respectively. The decrease in P solubility might result from the increased apatite P (increased by >85%) and decreased soluble Ca (decreased by ∼50%) after HTC. These results suggested that HTC could be an efficient strategy to immobilize P in cow manure, thereby potentially mitigating the P loss problem from cow manure.

Engineered hydrochar composites for phosphorus removal/recovery: Lanthanum doped hydrochar prepared by hydrothermal carbonization of lanthanum pretreated rice straw.

Engineered hydrochar composites (EHC) were synthesized by hydrothermal carbonization (HTC) of lanthanum pretreated rice straw. The as-prepared composite with about 30% lanthanum content showed greater P removal potential than La(OH)3, indicating the synergistic effect of hydrochar and lanthanum in P removal. The adsorption results showed that EHC showed great P adsorption capacities (>50mgPg(-1)) in the pH range of 2.5-10.5, and the presence of competing anions had little negative effects on P adsorption on EHC. The equilibrium time for P adsorption on EHC was considerably reduced under acid condition (12h) compared to alkaline condition (48h). The maximum adsorption capacity was 61.57mgPg(-1) according to Langmuir isotherms. These results suggested that EHC was highly effective in P adsorption in a wide range of pH and the presence of competing anions, thus EHC could be a promising adsorbent for phosphorus removal/recovery from wastewater.

Molecular cloning and characterization of the gene encoding squalene epoxidase in Panax notoginseng.

Squalene epoxidase (SE) is one of the rate-limiting enzymes in the triterpene saponins biosynthetic pathway. Panax notoginseng, one of the famous medicinal plants in China, produces bioactive triterpene saponins. Here we report the P. notoginseng SE, which was cloned from the root of P. notoginseng by PCR. The nucleotide sequence of the ORF (GenBank accession no. DQ386734) contains 1611 nucleotides and encodes 537 amino acid residues with molecular weight of 59.14 kDa and pI of 8.81. The gene has 98% identity with P. ginseng but different identities with other SE families. P. notoginseng SE has a FAD function domain, NAD(P)-binding Rossmann-fold domains, hydrophobicity and 4 transmembrane helices. This SE may be a microsomal membrane-associated enzyme. Real time quantitative PCR shows that the cDNA has different expression pattern and is highly expressed in root, especially in 3-year-old root.