Combining phospholipases and a liquid lipase for one-step biodiesel production using crude oils

Background Enzymatic biodiesel is becoming an increasingly popular topic in bioenergy literature because of its potential to overcome the problems posed by chemical processes. However, the high cost of the enzymatic process still remains the main drawback for its industrial application, mostly because of the high price of refined oils. Unfortunately, low cost substrates, such as crude soybean oil, often release a product that hardly accomplishes the final required biodiesel specifications and need an additional pretreatment for gums removal. In order to reduce costs and to make the enzymatic process more efficient, we developed an innovative system for enzymatic biodiesel production involving a combination of a lipase and two phospholipases. This allows performing the enzymatic degumming and transesterification in a single step, using crude soybean oil as feedstock, and converting part of the phospholipids into biodiesel. Since the two processes have never been studied together, an accurate analysis of the different reaction components and conditions was carried out. Results Crude soybean oil, used as low cost feedstock, is characterized by a high content of phospholipids (900 ppm of phosphorus). However, after the combined activity of different phospholipases and liquid lipase Callera Trans L, a complete transformation into fatty acid methyl esters (FAMEs >95%) and a good reduction of phosphorus (P <5 ppm) was achieved. The combination of enzymes allowed avoidance of the acid treatment required for gums removal, the consequent caustic neutralization, and the high temperature commonly used in degumming systems, making the overall process more eco-friendly and with higher yield. Once the conditions were established, the process was also tested with different vegetable oils with variable phosphorus contents. Conclusions Use of liquid lipase Callera Trans L in biodiesel production can provide numerous and sustainable benefits. Besides reducing the costs derived from enzyme immobilization, the lipase can be used in combination with other enzymes such as phospholipases for gums removal, thus allowing the use of much cheaper, non-refined oils. The possibility to perform degumming and transesterification in a single tank involves a great efficiency increase in the new era of enzymatic biodiesel production at industrial scale.

Screening for wound-induced oxylipins in Arabidopsis thaliana by differential HPLC-APCI/MS profiling of crude leaf extracts and subsequent characterisation by capillary-scale NMR

A simple non-targeted differential HPLC-APCI/MS approach has been developed in order to survey metabolome modifications that occur in the leaves of Arabidopsis thaliana following wound-induced stress. The wound-induced accumulation of metabolites, particularly oxylipins, was evaluated by HPLC-MS analysis of crude leaf extracts. A generic, rapid and reproducible pressure liquid extraction procedure was developed for the analysis of restricted leaf samples without the need for specific sample preparation. The presence of various oxylipins was determined by head-to-head comparison of the HPLC-MS data, filtered with a component detection algorithm, and automatically compared with the aid of software searching for small differences in similar HPLC-MS profiles. Repeatability was verified in several specimens belonging to different series. Wound-inducible jasmonates were efficiently highlighted by this non-targeted approach without the need for complex sample preparation as is the case for the 'oxylipin signature' procedure based on GC-MS. Furthermore this HPLC-MS screening technique allowed the isolation of induced compounds for further characterisation by capillary-scale NMR (CapNMR) after HPLC scale-up. In this paper, the screening method is described and applied to illustrate its potential for monitoring polar and non-polar stress-induced constituents as well as its use in combination with CapNMR for the structural assignment of wound-induced compounds of interest

Prediction of crude protein and classification of the growth stage of wheat plant samples from NIR spectra

Near-infrared spectroscopy (NIRS) was used to analyse the crude protein content of dried and milled samples of wheat and to discriminate samples according to their stage of growth. A calibration set of 72 samples from three growth stages of wheat (tillering, heading and harvest) and a validation set of 28 samples was collected for this purpose. Principal components analysis (PCA) of the calibration set discriminated groups of samples according to the growth stage of the wheat. Based on these differences, a classification procedure (SIMCA) showed a very accurate classification of the validation set samples : all of them were successfully classified in each group using this procedure when both the residual and the leverage were used in the classification criteria. Looking only at the residuals all the samples were also correctly classified except one of tillering stage that was assigned to both tillering and heading stages. Finally, the determination of the crude protein content of these samples was considered in two ways: building up a global model for all the growth stages, and building up local models for each stage, separately. The best prediction results for crude protein were obtained using a global model for samples in the two first growth stages (tillering and heading), and using a local model for the harvest stage samples.

In vitro antimicrobial and antioxidant activities of a crude extract and fractions from Buddleja thyrsoides Lam. Leaves

Crude extract and fractions of Buddleja thyrsoides were investigated regarding antioxidant activities by DPPH, total phenolic contents by Folin-Ciocalteau and antimicrobial activity by the broth microdilution method. Total phenolics varied from 214.07 ± 3.6 to 438.4 ± 0.3 mg g-1. Crude extract, ethyl acetate, dichloromethane and butanolic fractions exhibited a weak scavenging activity (SC50=186.04 ± 10.8, 137.70 ± 8.5, 146.89 ± 9.0 and 165.71 ± 3.2 µg mL-1, respectively). A correlation between the antioxidant activities and total phenolic contents could be shown (r=0.857, p<0.01). The lowest value of MIC was observed with butanolic fraction against Saccharomyces cerevisiae (MIC and MFC at 62.5 µg mL-1). Dichloromethane and ethyl acetate fractions were effective against Staphylococcus aureus with MIC value at 250 and 500 µg mL-1 respectively.

Effect of the structure of commercial poly(ethylene oxide-b-propylene oxide) demulsifier bases on the demulsification of water-in-crude oil emulsions: elucidation of the demulsification mechanism

Water-in-crude oil emulsions are formed during petroleum production and asphaltenes play an important role in their stabilization. Demulsifiers are added to destabilize such emulsions,however the demulsification mechanism is not completely known. In this paper, the performances of commercial poly(ethylene oxide-b-propylene oxide) demulsifiers were studied using synthetic water-in-oil emulsions and model-systems (asphaltenes in organic solvent). No change in the asphaltene aggregate size induced by the demulsifier was observed. The demulsification performance decreased as the asphaltene aggregate size increased, so it can be suggested that the demulsification mechanism is correlated to the voids between the aggregates adsorbed on the water droplets surface.

CHEMICAL STUDY OF Hortia superba (Rutaceae) AND INVESTIGATION OF THE ANTIMYCOBACTERIAL ACTIVITY OF CRUDE EXTRACTS AND CONSTITUENTS ISOLATED FROM Hortia SPECIES

In this paper, the chemical study of Hortia superba and antimycobacterial potential of Hortia species were investigated. Crude extracts and limonoids, alkaloids, dihydrocinnamic acid derivatives and coumarins isolated from Hortia superba, Hortia oreadica and Hortia brasiliana were evaluated against Mycobacterium tuberculosis H37Rv, Mycobacterium kansasii and Mycobacterium avium. The results obtained demonstrated an inhibitory effect of the dichloromethane extract of leaves of H. oreadica (MIC 31.25 µg mL-1), indolequinazoline (15.62 µg mL-1) and furoquinoline (31.25 µg mL-1) alkaloids, and dihydrocinnamic acid derivatives (62.50 µg mL-1), on the growth of M. tuberculosis. These results are promising in relation to the search for biologically active natural products and could be useful in the development of effective new drugs against mycobacteria.

Use of a Ca-selective electrode in the determination of total Ca in the production of crude sugar. Preliminary studies

The determination of the total calcium in juice, syrups, and other products of the sugar industry is investigated. Total calcium and free calcium is determinated by AAS and employing Ca-selective electrode respectively. A coefficient is obtained for the relation of total calcium with respect to free calcium. The coefficient is employed to determine the content of total calcium in accordance with the following equation.

Anaerobic co-digestion of crude glycerin and starch industry effluent

The Brazil's Biodiesel Production and Use Program introduces biodiesel in the Brazilian energy matrix, bringing along the perspective of a growth of the glycerin offer, co-product generated in the proportion of 10 kg for each 100 L of biodiesel. The aim of this study was to evaluate the addition of crude glycerin in the anaerobic digestion of cassava starch industry effluent (cassava wastewater), in a horizontal semi-continuous flow reactor of one phase in laboratory scale. It was used a reactor with a 8.77 L of useful volume, a medium support for corrugated conduit of polyvinyl chloride (PVC), temperature of 261 ºC, fed with cassava wastewater and glycerin, with hydraulic detention times of 4 and 5 days and increasing volumetric organic load of 3.05; 9.32; 14.83 and 13.59 g COD L-1 d-1, obtained with the addition of glycerin at 0; 2; 3 and 2% (v/v), respectively. The average removal efficiencies of TS and TVS were decreasing from the addition of glycerin to the cassava wastewater, averaging 81.19 to 55.58% for TS and 90.21 to 61.45% for TVS. The addition of glycerin at 2% increased the biogas production compared to the control treatment, reaching 1.979 L L-1 d-1. The biogas production as a function of the consumed COD was higher for the control treatment than for the treatments with addition of glycerin, which indicates lower conversion of organic matter into biogas.

In vitro and in vivo antitumor activity of crude extracts obtained from Brazilian Chromobacterium sp isolates

Natural products produced by microorganisms have been an important source of new substances and lead compounds for the pharmaceutical industry. Chromobacterium violaceum is a Gram-negative β-proteobacterium, abundant in water and soil in tropical and subtropical regions and it produces violacein, a pigment that has shown great pharmaceutical potential. Crude extracts of five Brazilian isolates of Chromobacterium sp (0.25, 2.5, 25, and 250 µg/mL) were evaluated in an in vitro antitumor activity assay with nine human tumor cells. Secondary metabolic profiles were analyzed by liquid chromatography and electrospray ionization mass spectrometry resulting in the identification of violacein in all extracts, whereas FK228 was detected only in EtCE 308 and EtCE 592 extracts. AcCE and EtCE 310 extracts showed selectivity for NCI/ADR-RES cells in the in vitro assay and were evaluated in vivo in the solid Ehrlich tumor model, resulting in 50.3 and 54.6% growth inhibition, respectively. The crude extracts of Chromobacterium sp isolates showed potential and selective antitumor activities for certain human tumor cells, making them a potential source of lead compounds. Furthermore, the results suggest that other compounds, in addition to violacein, deoxyviolacein and FK228, may be involved in the antitumor effect observed.

Effects of filtration methods on the neutralization yield of crude marine fish oil

The evaluation of filtration methods on the yield and oleochemicals characteristics on the crude fish oil from the soapstock of marine fish for purposes of nutrition has been conducted in this work. The analytical properties of the crude and the neutralized oil with three excesses of sodium hydroxide (NaOH, 20%, 40% and 60%) were carried out on two different ways with organza and the “glass wool”. The neutralization of the oil brought about a notable improvement in the analytical properties of the oil. Thus, it leads to a high quality fish oil in terms of taste, colour, odours, shelf life and market value. Based on the improved characteristics of the oil, it could be suitable for applications in pharmaceutical and food industries.

Effect of breed, gender, housing system and dietary crude protein content on performance of finishing beef cattle fed maize-silage-based diets

Maize silage-based diets with three dietary crude protein (CP) supplements were offered to 96 finishing cattle of contrasting breed (Holstein Friesian (HF) v. Simmental x HF (SHF)) and gender (bull v. steer) housed in two types of feeding system (group fed v. individually fed). The three protein supplements differed either in CP or protein degradability (degradable (LUDP) v. rumen undegradable (HUDP)) and provided CP concentrations of 142 (Con), 175 (LUDP) and 179 (HUDP) g/kg dry matter (DM) respectively, with ratios of degradable to undegradable of 3.0, 1.4 and 0.9:1 for diets Con, LOP and HUDP respectively. DM intakes were marginally higher (P = 0. 102) for LOP when compared with Con and HOP Rates of daily live-weight gain (DLWG) were higher (P = 0.005) in LUDP and HOP when compared with Con. HF had higher DM intakes than SHF although this did not result in any improvement in HF DLWG. Bulls had significantly better DM intakes, DLWG and feed conversion efficiency than steers. Conformation scores were better in SHF than HF (P < 0.001) and fat scores lower in bulls than steers (p < 0.001). There was a number of first order interactions established between dietary treatment, breed, gender and housing system with respect to rates of gain and carcass fat scores.

Antifeedant activities of crude seed extracts of tropical African spices against Spodoptera littoralis (Lepidoptera: Noctuidae)

The antifeedant activities of Piper guineense Schum et Thonn (Piperaceae), Aframomum melegueta (Rosk) K. Schum (Zingiberaceae), Aframomum citratum (Pareira) K. Schum (Zingiberaceae) and Afrostyrax kamerunensis Perkins and Gilg (Huaceae) seed extracts were investigated in laboratory dual- and no-choice bioassays using third-instar Spodoptera littoralis (Boisduval) larvae. In the dual-choice test, the hexane and methanol extracts of A. melegueta showed potent dose-dependent antifeedant activity at concentrations of ≥300 ppm and the water extract at ≥500 ppm, as illustrated by significantly lower leaf consumptions. Aframomum citratum methanol and water extracts exhibited antifeedant activity at ≥300 and ≥1000 ppm, respectively, but the hexane and ethanol extracts did not affect feeding at any concentration. Piper guineense ethanol and water extracts showed dose-dependent antifeedant effects at ≥300 and ≥500 ppm, respectively, and the methanol extract was active only at 1000 ppm. None of the extracts of the highly aromatic A. kamerunensis exhibited antifeedant activity at any of the tested concentrations. In the no-choice bioassays, extracts with antifeedant activity in the dual-choice tests also showed dose-dependent feeding inhibition. The hexane and methanol extracts of A. melegueta were effective in the no-choice tests at ≥100 and ≥500 ppm, respectively, and the water extract at ≥300 ppm. Similarly, the A. citratum water and methanol extracts were active at ≥500 ppm and the P. guineense water and ethanol extracts at ≥100 ppm. GC/MS chromatography of A. melegueta hexane and methanol extracts revealed volatile constituents with known anti-insect activity. The hexane and methanol extracts of A. melegueta, the methanol extract of A. citratum and the water and ethanol extracts of P. guineense may have potential for use by subsistence farmers.