DataSheet1_Decreasing acid value of fatty acid ethyl ester products using complex enzymes.PDF

Recently, enzymatic method has been used to prepare biodiesel using various oils. But the high acid value of the biodiesel product using enzyme as a catalyst has been one issue. In this work, an attempt to reduce the acid value of fatty acid ethyl ester (FAEE) product to satisfy the specified requirement (AV ≤ 0.5 mgKOH/g), a complex enzyme-catalyzed method was used for the ethanolysis of Semen Abutili seed oil (SASO) (AV = 5.5 ± 0.3 mgKOH/g). The effects of various variables (constituents of complex enzyme, type and addition of water removal agent, time, temperature, enzyme addition load, substrate ratio) on the enzymatic reaction were investigated. The optimal reaction conditions were: 1% addition of liquid lipase Eversa® Transform 2.0% and 0.8% of enzyme dry powder CALB, reaction temperature 35°C, alcohol-oil ratio 9:1 (mol/mol), 0.8 g/g of 4A-MS and reaction time 24 h. Under the optimal reaction conditions, the FAEE yield was 90.8% ± 1.5% and its acid value was decreased from 12.0 ± 0.2 mgKOH/g to 0.39 ± 0.10 mgKOH/g. In further evaluating the feasibility of preparing FAEE from SASO, the FAEE products obtained under the optimal reaction conditions were purified and evaluated with reference to the ASTM D6751 standard for the main physicochemical indexes. The results obtained were in accordance with the requirements except for the oxidative stability.</p

Sampling information and population statistics for <i>R</i>. <i>padi</i> investigated using <i>RpCPR</i>.

<p>Sampling information and population statistics for <i>R</i>. <i>padi</i> investigated using <i>RpCPR</i>.</p

Relative expression levels of <i>RpCPR</i> in the susceptible strain (SS), the isoprocarb-resistant strain (IS-R) and the imidacloprid-resistant strain (IM-R).

<p>The expression level of <i>RpCPR</i> in SS was set to 1. Data shown as the mean ± SE; different letters denoted a significant difference among samples (<i>P</i><0.05, one-way ANOVA).</p

Relative expression of <i>R</i>. <i>padi</i> CPR at different developmental stages.

<p>Relative expression levels of <i>RpCPR</i> at different developmental stages were normalized to those in the adult. Data shown as the mean ± SE; different letters denoted a significant difference among samples (<i>P</i><0.05, one-way ANOVA).</p

Molecular Cloning, Expression Pattern and Polymorphisms of NADPH-Cytochrome P450 Reductase in the Bird Cherry-Oat Aphid <i>Rhopalosiphum padi</i> (L.)

<div><p>NADPH–cytochrome P450 reductase (CPR) plays an important role in the cytochrome P450 (CYP)-mediated metabolism of endogenous and exogenous substrates. CPR has been found to be associated with insecticide metabolism and resistance in many insects. However, information regarding CPR in the bird cherry-oat aphid, <i>Rhopalosiphum padi</i>, is unavailable. In the current study, a full-length cDNA (2,476 bp) of <i>CPR</i> (<i>RpCPR</i>) encoding 681 amino acids was cloned from <i>R</i>. <i>padi</i>. Nucleotide sequence and deduced amino acid sequence analysis showed that RpCPR exhibits characteristics of classical CPRs and shares high identities with those of other insects, especially with the pea aphid, <i>Acyrthosiphon pisum</i>. The mRNA of <i>RpCPR</i> was expressed at all developmental stages, with the highest expression level found in the second instar and the lowest in adult. Expression levels of <i>RpCPR</i> in isoprocarb-resistant and imidacloprid-resistant strains were 3.74- and 3.53-fold higher, respectively, than that of a susceptible strain. <i>RpCPR</i> expression could also be induced by low concentrations (LC₃₀) of isoprocarb and imidacloprid. Moreover, we sequenced the open reading frame (ORF) of <i>RpCPR</i> from 167 field samples collected in 11 geographical populations. Three hundred and thirty-four SNPs were detected, of which, 65 were found in more than two individuals. One hundred and ninety-four missense mutations were present in the amino acid sequence, of which, the P484S mutant had an allele frequency of 35.1%. The present results suggest that <i>RpCPR</i> may play an important role in the P450-mediated insecticide resistance of <i>R</i>. <i>padi</i> to isoprocarb and imidacloprid and possibly other insecticides. Meanwhile, <i>RpCPR</i>maintains high genetic diversity in natural individuals, which provides the possibility of studying potential correlations between variants and certain special physiological characters.</p></div

<i>R</i>. <i>padi</i> CPR protein sequence depicting missense mutations or/and polymorphisms.

<p>Mutation found only in one individual is indicated by a hollow triangle while polymorphic variants identified in more than two individuals are indicated by a solid triangle. The digital number after the mutated amino acids indicated the number of the aphid individuals with the mutation.</p

Phylogenetic tree of <i>R</i>. <i>padi</i> CPR with other insect CPRs.

<p>The neighbor-joining tree was generated using MEGA 5.0 software, and the phylogeny was tested by the bootstrap method with 1,000 replications. Bootstrap values >50% are shown. <i>R</i>. <i>padi</i> CPR is indicated by solid circles. The GenBank accession numbers of the sequences used were listed in supporting information <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154633#pone.0154633.s001" target="_blank">S1 Table</a>.</p

Primers used for cloning and expression analysis of <i>RpCPR</i> in <i>R</i>. <i>padi</i>.

<p>Primers used for cloning and expression analysis of <i>RpCPR</i> in <i>R</i>. <i>padi</i>.</p

Gelatin Microgel Incorporated Poly(ethylene glycol)-Based Bioadhesive with Enhanced Adhesive Property and Bioactivity

Up to 7.5 wt % of chemically cross-linked gelatin microgel was incorporated into dopamine-modified poly­(ethylene glycol) (PEGDM) adhesive to simultaneously improve the material property and bioactivity of the PEG-based bioadhesive. Incorporation of gelatin microgel reduced cure time while it increased the elastic modulus and cross-linking density of the adhesive network. Most notably, the loss modulus values for microgel-containing adhesive were an order of magnitude higher when compared to microgel-free control. This drastic increase in the viscous dissipation ability of the adhesive is attributed to the introduction of reversible physical bonds into the adhesive network with the incorporation of the gelatin microgel. Additionally, incorporation of the microgel increased the adhesive properties of PEGDM by 1.5- to 2-fold. From in vitro cell culture studies, the composite adhesive is noncytotoxic and the incorporation of microgels provided binding site for promoting fibroblast attachment and viability. The subcutaneous implantation study indicated that the microgel-containing PEGDM adhesive is biocompatible and the incorporated microgels provided pockets for rapid cellular infiltration. Gelatin microgel incorporation was demonstrated to be a facile method to simultaneously enhance the adhesive property and the bioactivity of PEG-based adhesive

Sequence analysis of <i>R</i>. <i>padi</i> CPR.

<p>(A) Schematic drawing of RpCPR. Membrane anchor, conserved FMN-, FAD- and NADP-binding domains, FAD-binding motif (Arg₄₅₇ x Tyr₄₅₉ Ser₄₅₆), and catalytic residues (Ser₄₆₀, Cys₆₃₃, Asp₆₇₈ and Trp₆₈₀) are shown. (B) Comparison of the deduced amino acid sequence of RpCPR with those of other CPRs deposited into GenBank. The species and accession numbers are as follows: <i>Musca domestica</i> (NP_001273818), <i>Drosophila melanogaster</i> (NP_477158) and <i>Rattus norvegicus</i> (NP_113764).</p