Biotechnology Research and Innovation最新文献

After forty years of research development, an overview of solid-state fermentation (SSF), focusing on its applications, mainly of the very recent papers of the last five years, is presented. This review comprises the most important developed processes concerning the production of enzymes, biopulping processes, and traditional processes, for food fermentation, such as the production of Chinese daqu and koji, and industrial important biomolecules such as organic acids, pigments, phenolic compounds, aromas and biosorbents. SSF bioreactors design that has been developed is reported, so as the solutions for the classical drawbacks and the most important cases of successful employment of the technique are described. And, finally, it is summarized a very interesting report of patents and innovations regarding SSF products and processes.

The cotton boll weevil (Anthonomus grandis) is the most destructive cotton insect pest in Brazil. The endophytic habit of this insect makes difficult its chemical control. Chitin synthase (CHS) is an integral membrane glycosyltransferase that is essential for chitin chain polymerization and deposition in insect chitinous structures, such as the peritrophic membrane (PM). Because it is not present in plants or vertebrates, CHS can be considered a promising target for eco-friendly biotechnological approaches, such as RNA interference (RNAi)-mediated gene silencing. Considering the relevance of CHS genes in the chitin biosynthetic pathway in insects, we report here the molecular cloning of the full-length CHS2 cDNA from the cotton boll weevil, and its functional validation via RNAi. The AgraCHS2 cDNA sequence is 4,869 bp, with a 4,446 bp open reading frame that encodes a predicted protein with 1,482 amino acid residues. Predicted protein has high similarity (53 to 78%) with other insects CHS. Moreover, only one copy is present in A. grandis genome. Transcriptional analysis showed that AgraCHS2 transcripts are restricted to the insect midgut at the third-larval instar and adult stages, which are considered the main feeding stages. RNAi-mediated knockdown of the AgraCHS2 affected A. grandis development, resulting in oviposition reduction of 93% and leading to 100% adult mortality. These data, in addition to the observation of PM severe disorganization in the midgut after AgraCHS2 knock-down, suggest AgraCHS2 as a promising target for developing RNAi-based biotechnological alternatives to specifically control the cotton boll weevil.

In order to develop potent inhibitors of matrix metalloproteinase (MMP-2 and MMP-9) as anticancer agents, pharmacophore modeling and three-dimensional quantitative structure–activity relationship (3D-QSAR) models were established using PHASE 3.0. A pharmacophore 5-point (AAARR) model was developed for the studied dataset and the generated model was used to derive the predictive atom-based 3D-QSAR models. After identifying a valid hypothesis, we developed 3D-QSAR models applying the PLS algorithm. The selected 3D-QSAR models were suggestive of the vitality of the electron-withdrawing feature for the MMPs inhibitory potential. In addition, hydrophobic groups, hydrogen bond donor groups, positive ionic and negative ionic features also positively contributed to the MMPs inhibitory potential along with the electron-withdrawing feature. The developed models were statistically robust (MMP-2 Q² = 0.51; pred R² = 0.67; MMP-9 Q² = 0.59; pred R² = 0.77). The QSAR results help in identifying a relationship between structural features of hydroxamate derivatives and their activities which could be useful to design newer MMP inhibitors.

The immunization of animals has been carried out for centuries and is generally accepted as the most cost-effective and sustainable method of controlling infectious veterinary diseases. Up to twenty years ago, most veterinary vaccines were either inactivated organisms that were formulated with an oil-based adjuvant or live attenuated vaccines. In many cases, these formulations were not very effective. The discovery of antigen/gene delivery systems has facilitated the development of novel prophylactic and therapeutic veterinary vaccines. To identify vaccine candidates in genomic sequences, a revolutionary approach was established that stems from the assumption that antibodies are more readily able to access surface and secreted than cytoplasm proteins; as such, they represent ideal vaccine candidates. The approach, which is known as reverse vaccinology, uses several bioinformatics algorithms to predict antigen localization and it has been successfully applied to immunize against many veterinary diseases. This review examines some of the main topics that have emerged in the veterinary vaccine field with the use of modern biotechnology techniques.

Biomass consumption continues to increase worldwide for the provision of human energy needs. These high pressures for energy will determine the demand for crop plants as a resource for biofuel, heat and electricity. Thus, the search for plant traits associated with genetic increases in yield is unconditional. Here, we propose exploiting recent advances in plant biomass enhancement in non-crop as well as in crop plants. For this purpose, biotechnological approaches that are well known rapid ways of enhancing the plant traits, as well as the traditional way of improving plants through plant breeding selecting for desirable phenotypes are excellent techniques to improve plant biomass and reduce the dependence on fossil fuels. Obviously, many genes can be associated with promising phenotypes however this review will focus on genes selected from different plant networks.

Microorganisms are capable of converting insoluble phosphate into a bioavailable form through solubilization and mineralization processes. Hence in the present study a phosphate solubilizing bacterium, PSB-26, was isolated from mangrove of the Mahanadi delta using NBRIP-agar and NBRIP-BPB broth containing tricalcium phosphate as the phosphate source. Based on phenotypic and molecular characterization, the strain was identified as Alcaligenes faecalis. The maximum phosphate solubilizing activity of the strain was found to be 48 μg/ml with decrease in pH of the growth medium from 7.0 to 3.2. During phosphate solubilization, various organic acids, such as oxalic acid (289 mg/L), citric acid (0.2 mg/L), malic acid (0.3 mg/L), succinic acid (0.5 mg/L) and acetic acid (0.4 mg/L) produced in the broth culture were detected through HPLC analysis. Crude alkaline phosphatase activity of the strain was determined by p-nitrophenyl phosphate assay and optimized with different growth parameters to obtain maximum enzyme production. Under optimized sets of conditions, maximum alkaline phosphatase activity of 93.7 U/ml was observed. Partially purified alkaline phosphatase exhibited three protein bands of sizes approximately 45 kDa, 25 kDa and 17 kDa. Partially purified alkaline phosphatase during characterization showed maximum activity at pH 9.0 (96.53 U/ml), temperature of 45 °C (97.99 U/ml) and substrate concentration of 1.75 mg/ml (96.51 U/ml). The effect of the bacterium on growth of Arabidopsis thaliana plant showed that inoculation of bacterial culture exhibited better growth in comparison to the control. Hence the phosphate solubilizing and alkaline phosphatase production activity of the bacterium may have probable use for future biotechnological application.

Meloidogyne incognita is responsible for significant crop losses worldwide. The main strategy to control this phytopathogen is still based on synthetic nematicides that are harmful to human health and the environment. In this context, aqueous seed extracts of antagonistic plants were screened for molecules effective toward the infective stage (J₂) of M. incognita. The aqueous crude extract of Canavalia ensiformis (ACECe) showed the highest nematicidal activity (87 ± 3% mortality). ACECe dialysis fractionation allowed the collection of an external dialysate (EDCe) containing molecules smaller than 3.5 kDa effective against J₂ (96 ± 3.0% mortality); innocuous toward non targeted organisms as saprophytic nematodes, fungi, bacterium and insects larvae; active against J2 (96 ± 2% mortality) after heating (50 °C); no cytotoxic for bovine red blood cells; reduction of M. incognita eggs masses by 82.5% in tomato plants at green house conditions. Fractionation of the EDCe by reversed-phase high-performance liquid chromatography (RP-HPLC) separated five nematotoxic fractions. Analyses of those fractions based on gas chromatography–mass spectrometry (GC–MS), liquid chromatography–mass spectrometry (LCMS) and nuclear magnetic resonance (NMR) identified nine specific metabolites, follow-up testing of the individual authentic standards of each metabolite as the compounds: d-glucose, l-canavanine, xanthotoxin, cis-aconitic acid, trans-aconitic acid, malic acid, citric acid, palmitic acid and S-carboxymethylcysteine; all them confirmed to possess nematotoxic properties. However, some of those metabolites were not described previously demonstrating biological action against M. incognita.