Boosting Enzyme Activity in Enzyme Metal-Organic Framework Composites.

Enzymes, as highly efficient biocatalysts, excel in catalyzing diverse reactions with exceptional activity and selective properties under mild conditions. Nonetheless, their broad applications are hindered by their inherent fragility, including low thermal stability, limited pH tolerance, and sensitivity to organic solvents and denaturants. Encapsulating enzymes within metal-organic frameworks (MOFs) can protect them from denaturation in these harsh environments. However, this often leads to a compromised enzyme activity. In recent years, extensive research efforts have been dedicated to enhancing enzymatic activity within MOFs, leading to the development of new enzyme-MOF composites that not only preserve their catalytic potential but also outperform their free counterparts. This Review provides a comprehensive review on recent developments in enzyme-MOF composites with a specific emphasis on their enhanced enzymatic activity compared to free enzymes.

Alginate-based materials for enzyme encapsulation.

Enzymes are widely used in industry due to their high efficiency and selectivity. However, their low stability during certain industrial processes can result in a significant loss of catalytic activity. Encapsulation is a promising technique that can stabilize enzymes by protecting them from environmental stresses such as extreme temperature and pH, mechanical force, organic solvents, and proteases. Alginate and alginate-based materials have emerged as effective carriers for enzyme encapsulation due to their biocompatibility, biodegradability, and ability to form gel beads through ionic gelation. This review presents various alginate-based encapsulation systems for enzyme stabilization and explores their applications in different industries. We discuss the preparation methods of alginate encapsulated enzymes and analyze the release mechanisms of enzymes from alginate materials. Additionally, we summarize the characterization techniques used for enzyme-alginate composites. This review provides insights into the use of alginate encapsulation as a means of stabilizing enzymes and highlights the potential benefits for various industrial applications.

Improving phytase production in Pichia pastoris fermentations through de-repression and methanol induction optimization.

Pichia pastoris (Komagataella phaffii) is a fast-growing methylotrophic yeast with the ability to assimilate several carbon sources such as methanol, glucose, or glycerol. It has been shown to have outstanding secretion capability with a variety of heterologous proteins. In previous studies, we engineered P. pastoris to co-express Escherichia coli AppA phytase and the HAC1 transcriptional activator using a bidirectional promoter. Phytase production was characterized in shake flasks and did not reflect industrial conditions. In the present study, phytase expression was explored and optimized using instrumented fermenters in continuous and fed-batch modes. First, the production of phytase was investigated under glucose de-repression in continuous culture at three dilution factors, 0.5 d-1 , 1 d-1 , and 1.5 d-1 . The fermenter parameters of these cultures were used to inform a kinetic model in batch and fed-batch modes for growth and phytase production. The kinetic model developed aided to design the glucose-feeding profile of a fed-batch culture. Kinetic model simulations under glucose de-repression and fed-batch conditions identified optimal phytase productivity at the specific growth rate of 0.041 h-1 . Validation of the model simulation with experimental data confirmed the feasibility of the model to predict phytase production in our newly engineered strain. Methanol was used only to induce the expression of phytase at high cell densities. Our results showed that high phytase production required two stages, the first stage used glucose under de-repression conditions to generate biomass while expressing phytase, and stage two used methanol to induce phytase expression. The production of phytase was improved 3.5-fold by methanol induction compared to the expression with glucose alone under de-repression conditions to a final phytase activity of 12.65 MU/L. This final volumetric phytase production represented an approximate 36-fold change compared to the flask fermentations. Finally, the phytase protein produced was assayed to confirm its molecular weight, and pH and temperature profiles. This study highlights the importance of optimizing protein production in P. pastoris when using novel promoters and presents a general approach to performing bioprocess optimization in this important production host.

Encapsulation of enzymes in food industry using spray drying: recent advances and process scale-ups.

Enzymes are widely used in the food industry due to their ability in improving the functional, sensory, and nutritional properties of food products. However, their poor stability under harsh industrial conditions and their compromised shelf-lives during long-term storage limit their applications. This review introduces typical enzymes and their functionality in the food industry and demonstrates spray drying as a promising approach for enzyme encapsulation. Recent studies on encapsulation of enzymes in the food industry using spray drying and the key achievements are summarized. The latest developments including the novel design of spray drying chambers, nozzle atomizers and advanced spray drying techniques are also analyzed and discussed in depth. In addition, the scale-up pathways connecting laboratory scale trials and industrial scale productions are illustrated, as most of the current studies have been limited to lab-scales. Enzyme encapsulation using spray drying is a versatile strategy to improve enzyme stability in an economical and industrial viable way. Various nozzle atomizers and drying chambers have recently been developed to increase process efficiency and product quality. A comprehensive understanding of the complex droplet-to-particle transformations during the drying process would be beneficial for both process optimization and scale-up design.

Encapsulation of enzyme using spray drying is a versatile approach for improving enzyme stability and shelf-life in food industry.This paper gives an overview of recent development and progress in enzyme encapsulation using spray drying.Emerging spray drying techniques and novel design of spray drying chambers and atomizers are summarized.Ex ante process simulations and technoeconomic analysis are also presented providing critical insights for commercial production of encapsulated enzymes.

Linking the neural basis of distributional statistical learning with transitional statistical learning: The paradox of attention.

Statistical learning, the process of tracking distributional information and discovering embedded patterns, is traditionally regarded as a form of implicit learning. However, recent studies proposed that both implicit (attention-independent) and explicit (attention-dependent) learning systems are involved in statistical learning. To understand the role of attention in statistical learning, the current study investigates the cortical processing of distributional patterns in speech across local and global contexts. We then ask how these cortical responses relate to statistical learning behavior in a word segmentation task. We found Event-Related Potential (ERP) evidence of pre-attentive processing of both the local (mismatching negativity) and global distributional information (late discriminative negativity). However, as speech elements became less frequent and more surprising, some participants showed an involuntary attentional shift, reflected in a P3a response. Individuals who displayed attentive neural tracking of distributional information showed faster learning in a speech statistical learning task. These results suggest that an involuntary attentional shift might play a facilitatory, but not essential, role in statistical learning.

Alginate Particles for Enzyme Immobilization Using Spray Drying.

Enzymes are important catalysts for biological processes due to their high catalytic activity and selectivity. However, their low thermal stability limited their industrial applications. The present work demonstrates a simple and effective method for enzyme immobilization via spray drying. Alginate was used as a support material. Phytase, an important enzyme in the animal feed industry, was selected to study the effect of enzyme immobilization using alginate particles on its thermal stability. The physicochemical properties of alginate particles such as size, surface morphology, and heat resistance were studied. Successful immobilization of phytase was confirmed by confocal microscopy, and the immobilized phytase retained 58% of its original activity upon heating at 95 °C, compared to 4% when the alginate support material was absent. Phytase was released promptly in a simulated gastrointestinal tract with >95% of its original activity recovered. The spray drying method for phytase immobilization is scalable and applicable to other enzymes for various applications.

Reduced right-hemisphere ERP P600 grammaticality effect is associated with greater right-hemisphere inhibition: Evidence from right-handers with familial sinistrality.

The present study examined the hypothesis that left hemisphere (LH) equivalent language capabilities in the right hemisphere (RH) are inhibited in neurologically intact individuals by testing healthy young right-handers with a history of familial sinistrality (FS+, i.e. with at least one left handed biological relative), a population documented to show greater variability for RH language processing. Event-Related Potential (ERP) and split visual field presentation techniques were combined to assess LH- and RH- biased responses to syntactic category violations. In addition, a bilateral flanker task was used to measure inter-hemispheric inhibition ability in the same set of participants. Replicating prior findings, in addition to the LH-biased P600 grammaticality effect previously seen for right-handers in general, a fair amount, though not all, of FS + right-handers showed RH-biased P600 responses, leading to a reliable RH P600 grammaticality effect at the group level. Capitalizing on the variability of RH P600 responses, our results further revealed that reduced RH-biased P600 effects were reliably correlated with more effective RH inhibition (indexed by smaller reaction time differences between incongruent and neutral flankers presented to the RH via the left visual field). These results corroborated previous findings that the RH is capable of processing syntactic information in a manner qualitatively similar to that in the LH and further demonstrated that LH-equivalent processing in the RH as indexed by the P600 responses is modulated by RH inhibition, contributing to inter-individual variability in syntactic lateralization.