Technological Implications of Modifying the Extent of Cell Wall-Proanthocyanidin Interactions Using Enzymes

The transference and reactivity of proanthocyanidins is an important issue that affects the technological processing of some fruits, such as grapes and apples. These processes are affected by proanthocyanidins bound to cell wall polysaccharides, which are present in high concentrations during the processing of the fruits. Therefore, the effective extraction of proanthocyanidins from fruits to their juices or derived products will depend on the ability to manage these associations, and, in this respect, enzymes that degrade these polysaccharides could play an important role. The main objective of this work was to test the role of pure hydrolytic enzymes (polygalacturonase and cellulose) and a commercial enzyme containing these two activities on the extent of proanthocyanidin-cell wall interactions. The results showed that the modification promoted by enzymes reduced the amount of proanthocyanidins adsorbed to cell walls since they contributed to the degradation and release of the cell wall polysaccharides, which diffused into the model solution. Some of these released polysaccharides also presented some reactivity towards the proanthocyanidins present in a model solution.     

Effect of a cold‐active pectinolytic system on colour development of Malbec red wines elaborated at low temperature

The effect of a new cold‐active pectinolytic system on colour of Malbec wines was studied under the following winemaking conditions: (i) fermentation at low temperature (20 °C) and (ii) prefermentative cold maceration (PCM) (5 °C–7 days) followed by traditional fermentation (28 °C). The pectinolytic system was mainly composed of polymethylgalacturonase and pectin lyase activities, detected under similar conditions to those in winemaking (pH 3.6–20 °C). The results show that the enzyme system significantly accelerated colour extraction by reducing the maceration time necessary for vinification at low temperature and shortening the PCM stage. Enzyme‐treated wines exhibited better chromatic parameters than their controls at devatting and after 6 months of storage. The cold‐active enzyme compensated the decrease in colour extraction due to the low maceration temperature, achieving high‐quality wines with chromatic characteristics similar to those of traditional wines.     

A study of external mass transfer effect on biodegradation of phenol using low‐density polyethylene immobilized Bacillus flexus GS1 IIT (BHU) in a packed bed bioreactor

The impact of external mass transport on the biodegradation rate of phenol in a packed bed bioreactor (PBBR) was studied. A potential bacterial species, Bacillus flexus GS1 IIT (BHU), was isolated from the petroleum‐contaminated soil. Low‐density polyethylene (LDPE) immobilized with the B. flexus GS1 IIT (BHU) was used as packing material in the PBBR. The PBBR was operated by varying the inlet feed flow rate from 4 to 10 mL/min, and the corresponding degradation rate coefficients were found to be in the range of 0.119–0.157 L/g h. In addition, the highest removal rate of phenol was obtained to be 1.305 mg/g h at an inlet feed rate of 10 mL/min. The external mass transfer was studied using the model . A new empirical correlation for the biodegradation of phenol in the PBBR was developed after the evaluation at various values of K and n.     

From Active Sites to Machines: A Challenge for Enzyme Chemists

As researchers who study enzyme chemistry embrace increasingly complex systems, especially biological machines, our attention is also shifting from steps involving covalent bond formation or cleavage to those that exclusively involve changes in non‐covalent bonding. Assembly line polyketide synthases are an example of this growing challenge. By now, the chemical reactions underpinning polyketide biosynthesis can be unequivocally mapped to well‐defined active sites and are, for the most part, readily explicable in the language of physical organic chemistry. Yet, all of these insights merely serve as a backdrop to the real problem of explaining how the catalytic functions of dozens of active sites are synchronized in order to allow these remarkable machines to turn over with remarkable specificity. Notwithstanding the fact that the time‐honored language of physical organic chemistry can teach us a lot, it is often insufficient to describe many of these events, and must therefore evolve.     

Resonance Assignments of Lowly Populated and Unstable Enzyme Intermediate Complex under Real-Time Conditions

Unstable and low-abundance protein complexes represent a large family of transient protein complexes that are difficult to characterize, even by means of high-resolution NMR spectroscopy. A method to assign the NMR signals of these unstable complexes through a combination of selective isotope labeling of amino acids in a protein and site-specific labeling the protein with a paramagnetic tag is presented herein. By using this method, the resonances of unstable thioester intermediate complex (lifetime <5 h and highest concentration ≈20 μm ) generated by Staphylococcus aureus sortase A and its peptide substrate under a real-time reaction have been assigned.     

Improved microstructure and properties of the gelatin film produced through prior cross‐linking induced by horseradish peroxidase,glucose oxidase and glucose

The effects of prior enzymatic cross‐linking of bovine gelatin via horseradish peroxidase, glucose oxidase and glucose on microstructure and properties of the target film (cross‐linked gelatin film) were assessed. The cross‐linked gelatin film exhibited similar film thickness and moisture content, lower water solubility and higher opacity than the gelatin film directly prepared with bovine gelatin. The cross‐linked gelatin film also demonstrated improved thermal stability and mechanical properties, characterised by higher melting point and glass transition temperature, enhanced tensile strength and elongation at break and greater storage modulus. Prior gelatin cross‐linking resulted in 30.2% and 68.6% reductions in water vapour and CO₂ permeability of the cross‐linked gelatin film, respectively, but did not affect oil permeability. Furthermore, the cross‐linked gelatin film possessed smaller cross‐sectional voids (diameter 100?360 nm vs. 200?595 nm) than the bovine gelatin film. This study shows that cross‐linking can efficiently improve film microstructure and properties of the gelatin‐like products.     

Cell-envelope proteinases from lactic acid bacteria: Biochemical features and biotechnological applications

Proteins displayed on the cell surface of lactic acid bacteria (LAB) perform diverse and important biochemical roles. Among these, the cell-envelope proteinases (CEPs) are one of the most widely studied and most exploited for biotechnological applications. CEPs are important players in the proteolytic system of LAB, because they are required by LAB to degrade proteins in the growth media into peptides and/or amino acids required for the nitrogen nutrition of LAB. The most important area of application of CEPs is therefore in protein hydrolysis, especially in dairy products. Also, the physical location of CEPs (i.e., being cell-envelope anchored) allows for relatively easy downstream processing (e.g., extraction) of CEPs. This review describes the biochemical features and organization of CEPs and how this fits them for the purpose of protein hydrolysis. It begins with a focus on the genetic organization and expression of CEPs. The catalytic behavior and cleavage specificities of CEPs from various LAB are also discussed. Following this, the extraction and purification of most CEPs reported to date is described. The industrial applications of CEPs in food technology, health promotion, as well as in the growing area of water purification are discussed. Techniques for improving the production and catalytic efficiency of CEPs are also given an important place in this review.