Effect of malting process duration on malting losses and quality of wheat malts

The study assesses impact of malting process duration on malting losses and quality of malts obtained from three varieties of winter wheat, i.e., Elixer, Rockefeller and Gimantis. The findings show that increased duration of the malting process (from 4 to 7 days) corresponded to greater total weight loss, with the most significant differences observed between 5-day and 6-day wheat malts. The qualitative analysis of the malts was carried out in accordance with EBC methodology. The assessments showed that the 5-day long malting process applied to the relevant wheat varieties resulted in production of high-quality malt with optimum malting losses observed in the case of Elixer and Rockefeller varieties. Slightly higher malting losses were identified in the case of Gimantis, and the malt obtained from this variety had very high contents of soluble protein (on average 5.34% d.m.) and Kolbach Index (average of 50.49%), which reflects high proteolytic activity during the grain malting process and a need to modify the malting process for this variety.

Interactome analyses reveal profound proteasome structural and functional rearrangements throughout mammalian spermatogenesis

During spermatogenesis, spermatogonia undergo a series of mitotic and meiotic divisions on their path to spermatozoa. To achieve this, a succession of complex processes requiring high proteolytic activity are in part orchestrated by the proteasome. The spermatoproteasome (s20S) is a proteasome subtype specific to the developing gametes, in which the gamete-specific α4s subunit replaces the α4 isoform found in the constitutive proteasome (c20S). Although the s20S is conserved across species and was shown to be crucial for germ cell development, its mechanism, function and structure remain incompletely characterized. Here, we used advanced mass spectrometry (MS) methods to map the composition of proteasome complexes and their interactomes throughout spermatogenesis. We observed that the s20S becomes highly activated as germ cells enter meiosis, mainly through association with proteasome activators PA200 and 19S. Additionally, the proteasome population shifts from predominantly c20S (98%) to predominantly s20S (>82-92%) during differentiation, presumably due to the shift from α4 to α4s expression. We confirmed that s20S, but not c20S, interacts with components of the synaptonemal complex, the multi-protein assembly that connects homologous chromosomes during meiosis. In vitro, s20S preferentially bind to 19S, and displayed higher trypsin- and chymotrypsin-like activities, both with and without PA200 activation. Moreover, using MS methods to monitor protein dynamics, we identified significant differences in domain flexibility between α4 and α4s. We propose that these differences induced by α4s incorporation result in significant changes in the way the s20S interacts with its partners, and dictate its role in germ cell differentiation.

Technological Properties and Biogenic Amines Production by Bacteriocinogenic Lactococci and Enterococci Strains Isolated from Raw Goat's Milk

ABSTRACT Technological properties and biogenic amine production were analyzed in 56 bacteriocinogenic lactococci and enterococci strains isolated from raw goat's milk. Fifteen lactococci strains were able to reduce milk pH to 5.3 or lower after 6 h, while enterococci strains were initially slow in producing acids. Lactococcus lactis subsp. lactis GLc06 and three strains of Enterococcus faecalis (GEn20, GEn22, and GEn23) presented high proteolytic activity. L. lactis subsp. lactis GLc06 and E. faecalis GEn22 also showed a high percentage of autolysis after only 4 h, reaching 71.11 and 97.67%, respectively, after 24 h. No strain was able to secrete exopolysaccharides, and L. lactis subsp. lactis GLc22 and 25 of the Enterococcus strains were able to produce diacetyl. L. lactis subsp. lactis GLc05 and 23 of the Enterococcus strains presented a high tolerance to NaCl at 10% (wt/vol). Regarding biogenic amine production, 12 strains (5 lactococci and 7 enterococci) were capable of forming tyramine and 4 strains (1 lactococcus and 3 enterococci) were capable of forming 2-phenylethylamine, but in very low amounts. GLc06 presented great acidifying, proteolytic, and autolytic activities, and GLc05 was capable of growing at high NaCl concentrations (10%, wt/vol), possessing medium autolytic and proteolytic activities. Some enterococci strains produced diacetyl and high autolytic and extracellular proteolytic activities and also presented resistance to high NaCl concentrations. The interesting technological properties presented by some bacteriocinogenic strains can justify their use by the dairy industry, with the aim of ensuring both safety due to bacteriocin production and technological transformations in fermented products.

Featured Article: Nanoenhanced matrix metalloproteinase-responsive delivery vehicles for disease resolution and imaging

The wide array of proteases, including matrix metalloproteinases, produced in response to many pathogenic insults, confers a unique proteolytic signature which is often disease specific and provides a potential therapeutic target for drug delivery. Here we propose the use of collagen-based nanoenhanced matrix metalloproteinase-responsive delivery vehicles that display matrix metalloproteinase-specific degradation in diverse in vitro models of proteolysis. We demonstrate that collagen particles comprised of protease substrates (primarily collagen) can be made of uniform size and loaded efficiently with assorted cargo including fluorescently labeled mesoporous silica, magnetic nanoparticles, proteins and antioxidants. We also demonstrate that pathologic concentrations of proteases produced in situ or in vitro display protease-specific cargo release. Additionally, we show that the collagen-based particles display bright fluorescence when loaded with a fluorophore, and have the potential to be used as vehicles for targeted delivery of drugs or imaging agents to regions of high proteolytic activity.

Proteolytic Activity in the Midgut of the Crimson Speckled Moth Utethesia Pulchella L. (Lepidoptera: Arctiidae)

Proteolytic Activity in the Midgut of the Crimson Speckled MothUtethesia PulchellaL. (Lepidoptera: Arctiidae)Samples were prepared from the midgut of 4th instar larvae of the crimson speckled mothUtethesia pulchellaL. to find proteolytic activity and properties. Result revealed the presence of high proteolytic activity in the midgut when taking into account specific proteinases including trypsin-like, chymotrypsin-like, elastase and two exopeptidase (aminopeptidase and carboxipeptidase). The optimal pH of general protease was 8 and 7 when using azocasein and hemoglobin as general substrates, respectively. The optimal temperature of the total proteolytic activity in the midgut ofU. pulchellawas 25°C and 30°C when using azocasein and hemoglobin, respectively. Proteolytic activity was inhibited significantly by soybean trypsin inhibitor (SBTI), phenylmethylsulfonyl fluoride (PMSF), trypsin inhibitor (TLCK), chymotrypsin inhibitor (TPCK) and Phenanthroline. These results provide evidences for the presence of serine proteinases as the major proteases in the midgut ofU. pulchella;a key rangeland pest in warm climates. The interaction between digestive proteases and protease inhibitors have potentially important consequences for pest management programs.

Occurrence and identification of yeasts in dogs external ear canal with and without otitis

ABSTRACTObjective. To analyze the presence of yeast in the external ear canal of 116 dogs with and without a diagnosis of otitis from veterinary clinic in the Chapecó city, Santa Catarina, Brazil, and to examine the secretion of the proteinase in isolates. Materials and methods. Were collected cerumen of conduct hearing of dogs of 16 different races 71% with pendular ear type, 5% of semi-pendular and 24% of the erect type. All dogs were previously evaluated by otoscopy and grouped in dogs with and without otitis. Results. Yeasts were isolated in 44 samples (approximately 36%), where Malassezia pachydermatis was identified in 95% of samples where were observed growth of yeasts. On 20 samples the proteinase enzyme showed strong activity in 31% isolates, were 21% of the dogs with otitis tested showed high proteolytic activity. Conclusions. We observed a variation of strains of M. pachydermatis-producing enzymes. The variation in production of these enzymes is probably more associated with different response to the action of the immune system of the animal in the tissue injury.

Shelf life extension of liquid whole eggs by heat and bacteriocin treatment

During a 15-month period, samples of commercially pasteurised liquid whole egg (LWE) were tested for the presence of spoilage microflora. The total bacterial counts were 2.2 ± 0.6 log CFU/g and total lactic acid bacteria (LAB) counts were 1.9 ± 0.6 log CFU/g. Enterobacteriaceae were detected in 2 samples. Out of the tested samples, 45 LAB were isolated and identified, with 30 strains identified as Enterococcus faecium, 12 as Enterococcus faecalis, and 3 as Lactobacillus paracasei subsp. paracasei. All strains, except 6 strains of E. faecium, possessed lipolytic activity. All the E. faecalis strains and one strain of E. faecium showed a high proteolytic activity, while moderate proteolytic activity was shown by 3 lactobacilli strains. Minimum inhibitory concentration (MIC) of nisin and Micocin X was measured against groups of isolated strains, and ranged from 10.4 µg/ml to 41.7 µg/ml for nisin and from 0.2 mg/ml to 1.6 mg/ml for Micocin X. The LWEs supplemented with 6.25 mg/l of nisin or with 500 mg/ml of Micocin X were pasteurised at 65°C for 2.5 minutes. The shelf life of LWE with the addition of nisin or Micocin X stored under refrigerator conditions was extended by a minimum of 5 weeks.

Exploring possibilities for quality improvement of meat raw materials from cattle ruminant animals by enzymatic treatment

The production of beef meat is a widely spread in Republic of Macedonia, and the beef is very abundant with binding tissue that influences certain hardness and tenacity both to the meat and the final meat product, thus diminishing its organoleptic quality. That demands for seeking adequate methods to improve the consistency and succulence of meat. For that purpose, were used a proteolytic enzyme preparations derived from the bacteria Streptomyces species 82 which have a high proteolytic activity. In this study was determined increased solubility of myofibril proteins at the meat (Muscles supraspinatus) and beef hearts when treated with enzyme preparations. Both Muscles supraspinatus and beef hearts showed a general tendency to increase the quantity of free amino acids when an enzyme preparation with activity of 110 PU/kg was added.