Improving of Gelling Capacity of Cooked Crab Meat Proteins

Cooked crab meat subjected to a cutting process can aggregate again, forming weak gels. The objective of this work was to determine the effect of two mixing methods, combined with the addition of the microbial enzyme TGase (MTGase) on the mechanical and functional properties of gels from washed or unwashed blue crab (Callinectes sapidus) meat. Live crabs were obtained from Laguna Madre, Tamaulipas, Mexico, and cooked at 120°C for 20 min before hand-picking the meat from the shell. Cooked meat was processed by mixing and cut at temperatures of 25 or 60°C, without (control) or 0.5% of MTGase. Then cooked at 90°C for 15 min. Changes in texture profile analysis, percentage of extractable water, and color were evaluated. The mixing method at 60°C allowed increasing the textural properties of the gels, and the addition of MTGase significantly improved the mechanical properties. The results allowed stablishing a viable technique to obtain restructured gels from cooked crab meat with no need to extract the soluble compounds responsible for their distinctive odor and taste which often affect the mechanical properties.

PcGCE is a potent elicitor of defense responses in aspen

Using microbial enzymes in transgenesis is a powerful means to introduce new functionalities in plants. Glucuronoyl esterase (GCE) is a microbial enzyme hydrolyzing the ester bond between lignin and 4-O-methyl-α-D-glucuronic acid present as a side chain of glucuronoxylan. This bond mediates lignin-carbohydrate complex (LCC) formation, considered as crucial factor of lignocellulose recalcitrance to saccharification. Previous studies showed that hybrid aspen (Populus tremula L. x tremuloides Michx.) constitutively expressing Phanerochaete carnosa Burt GCE (PcGCE) had better efficiency of cellulose-to-glucose conversion but were stunned and had lower cellulose content indicating that more studies are needed to design strategy for deployment of this enzyme in planta. Here we report that the transgenic plants exhibit premature leaf senescence, increased accumulation of calcium oxalate crystals, tyloses and necrotic lesions and have strongly activated immune defense reactions as revealed by their altered profiles of transcriptomes, metabolomes and hormones in the leaves. To elucidate if these effects are triggered by damage-associated molecular patterns (DAMPs) or by PcGCE protein perceived as a pathogen-associated molecular pattern (PAMP), we ectopically expressed in aspen an enzymatically inactive PcGCES217A. The mutated PcGCE induced similar growth retardation, leaf necrosis and premature senescence as the active one, providing evidence that PcGCE protein is recognized as PAMP. Transcriptomics analysis of young expanding leaves of 35S:PcGCE plants identified several candidates for receptors of PcGCE, which were not expressed in developing wood tissues. Grafting experiments showed that PcGCE transcripts are not cell-to-cell mobile and that leaves augment systemic responses. In agreement, expressing PcGCE in developing wood by using the wood-specific promoter (WP), avoided all off-target effects. Moreover, WP:PcGCE lines grew better than control plants providing evidence that this strategy can be used in transgenic crops dedicated for biorefinery.

A brief review on conventional methods for screening, product development, downstream and evaluation of fibrinolytic enzymes from marine microbes

The ocean is a great reservoir of biodiversity and microbial metabolites. Enzymes from marine source have recently gained considerable attention as they have lower side effects and more potency when compared to other existing sources. Fibrinolytic enzymes from microbial sources possess ability to dissolve clots and help to circumvent cardiovascular problems in more efficient and safer way. The complexity of the marine environment involves high salinity, high pressure, low temperature, special lighting conditions. This contributes to the significant differences between the enzymes generated by marine microorganisms and homologous enzymes from terrestrial microorganisms leading to the boosted marine microbial enzyme technology. Further, it is believed that sea water, which is saline in nature and chemically closer to the human blood plasma, could provide biomolecules, in particular enzymes that could have lower or no toxicity or side effects when used for therapeutic applications. However, only a small proportion of fibrinolytic enzymes from marine microbiota has been examined and an even smaller proportion has been exploited. Therefore, much work needs to be done intensively and extensively in terms of potent fibrinolytic enzymes from marine resources.

Functional Diversity of Morphologically Similar Digestive Organs in Muroidea Species

We examine possible ways of functional adjustment of morphologically similar alimentary tracts in rodents with different dietary specializations. We study the structure of stomach and gut epithelial surface as well as the features of its colonization with microorganisms in five gerbil species: Psammomys obesus, Meriones crassus, Gerbillus henleyi, G. andersoni, and G. dasyurus. Data on the morphological diversity of mucosa-associated microbiota have been obtained and confirmed by the results of previous microbiology studies. Species differences in chymus acidity associated with dietary specialization have been determined. Variations in the activity of the endoglucanase microbial enzyme, which is crucial for rodents fed on cellulose-containing food, have also been detected. The importance of microbiota for functional adaptations to various food types in rodents with morphologically similar digestive tracts has been evaluated.

An enzyme-based biosensor for monitoring and engineering protein stability in vivo

Protein stability affects the physiological functions of proteins and is also a desirable trait in many protein engineering tasks, yet improving protein stability is challenging because of limitations in methods for directly monitoring protein stability in cells. Here, we report an in vivo stability biosensor wherein a protein of interest (POI) is inserted into a microbial enzyme (CysGA) that catalyzes the formation of endogenous fluorescent compounds, thereby coupling POI stability to simple fluorescence readouts. We demonstrate the utility of the biosensor in directed evolution to obtain stabilized, less aggregation-prone variants of two POIs (including nonamyloidogenic variants of human islet amyloid polypeptide). Beyond engineering applications, we exploited our biosensor in deep mutational scanning for experimental delineation of the stability-related contributions of all residues throughout the catalytic domain of a histone H3K4 methyltransferase, thereby revealing its scientifically informative stability landscape. Thus, our highly accessible method for in vivo monitoring of the stability of diverse proteins will facilitate both basic research and applied protein engineering efforts.