Synthesis of Long-Chain β-Lactones and Their Antibacterial Activities against Pathogenic Mycobacteria

In the quest for new antibacterial agents, a series of novel long- and medium-chain mono- and disubstituted β-lactones was developed. Their activity against three pathogenic mycobacteria—M. abscessus, M. marinum, and M. tuberculosis—was assessed by the resazurin microtiter assay (REMA). Among the 16 β-lactones synthesized, only 3-hexadecyloxetan-2-one (VM005) exhibited promising activity against M. abscessus, whereas most of the β-lactones showed interesting activities against M. marinum, similar to that of the classical antibiotic, isoniazid. Regarding M. tuberculosis, six compounds were found to be active against this mycobacterium, with β-lactone VM008 [trans-(Z)-3-(hexadec-7-en-1-yl)-4-propyloxetan-2-one] being the best growth inhibitor. The promising antibacterial activities of the best compounds in this series suggest that these molecules may serve as leads for the development of much more efficient antimycobacterial agents.     

Effect of advanced chilling methods on lipid damage during sardine (Sardina pilchardus) storage

Slurry ice is a biphasic system consisting of small spherical ice crystals surrounded by seawater at subzero temperature. Its effect on lipid damage (hydrolysis and oxidation) was evaluated during the chilled storage of a fatty fish species, sardine (Sardina pilchardus). Slurry ice treatment was checked alone and in combination with ozone and compared to traditional flake icing during a 22‐day storage. Different lipid damage indices (free fatty acids, FFA; peroxide value, PV; thiobarbituric acid index, TBA‐i; fluorescent compounds, FR) were checked and compared to sensory assessment and nucleotide degradation (K value). According to lipid hydrolysis (FFA) and oxidation (PV and FR) developments, slurry ice showed an inhibitory effect (p <0.05) on lipid damage during storage, as well as an inhibition of nucleotide autolytic degradation. Ozonised slurry ice did not provide differences (p >0.05) from slurry ice alone when considering lipid hydrolysis, nucleotide degradation and some lipid oxidation indices (PV and FR), although a higher (p <0.05) TBA‐i was observed at day 22 of storage when compared to flake ice and slurry ice treatments. However, a lower (p <0.05) fluorescence development was observed for fish treated under ozonised slurry ice when compared to traditionally iced fish. Sensory assessment showed a higher shelf life for fish samples treated under ozonised slurry ice than for their counterparts under slurry ice (15 d versus 12 d), while flake icing led to a far shorter shelf life (5 d). According to sensory and biochemical (lipid matter and nucleotide) analysis, slurry ice has proved to be a promising technology for damage inhibition and quality retention in a fatty fish species such as sardine. Ozonised slurry ice was also shown to be useful, since a longer shelf life was obtained in the present experiment and a pro‐oxidant effect of ozone on sardine lipids was not proved.     

产高温蛋白酶高温菌的筛选

利用平板透明圈法，从堆肥、土壤中筛选到88种产高温蛋白酶的高温菌，它们在酪素筛选平板上都产生明显的蛋白水解透明圈。这些产高温蛋白酶高温菌株的获得为进一步研究高温菌的耐热机制、高温酶的热稳定性机理等理论问题及高温蛋白酶的应用，奠定了基础。     

Hydrolytic degree and antioxidant activity of purified casein characterised by different haplotypes (αs1-, β- and k-casein) after enzymatic hydrolysis with pepsin and enzymatic extract from Pleurotus eryngii

The aim of this study was to estimate the hydrolytic degree and antioxidant activity of purified casein characterised by different haplotypes (α_s1-, β- and k-casein) after in vitro digestion with two different enzymatic systems: pepsin from porcine gastric mucosa (EP) and a crude enzymatic extract from the edible mushroom Pleurotus eryngii. The used enzymes showed a different mode of casein catalysis with a consequent production of peptides of different antioxidant activity. The CN haplotype significantly influenced peptides production; in fact, the amino acid substitutions caused by genetic polymorphisms at the α_s1-, β- and k-CN loci influenced the sites of enzymatic cleavage and therefore the produced peptides. The above is evidenced by the different antioxidant activity found in the hydrolysates depending on the used enzymatic system, the CN haplotype, and the CN haplotype × enzymatic treatment interaction. The findings of this study are a perspective for the production of specific foods that exert a biological effect in addition to the nutritional one.     

Sugarcane Serine Peptidase Inhibitors,Serine Peptidases,and Clp Protease System Subunits Associated with Sugarcane Borer (Diatraea saccharalis) Herbivory and Wounding

Sugarcane’s (Saccharum spp.) response to Diatraea saccharalis (F.) (Lepidoptera: (Crambidae) herbivory was investigated using a macroarray spotted with 248 sugarcane Expressed Sequence Tags (ESTs) encoding serine peptidase inhibitors, serine peptidases. and Clp protease system subunits. Our results showed that after nine hours of herbivory, 13 sugarcane genes were upregulated and nine were downregulated. Among the upregulated genes, nine were similar to serine peptidase inhibitors and four were similar to Bowman-Birk Inhibitors (BBIs). Phylogenetic analysis revealed that these sequences belong to a phylogenetic group of sugarcane BBIs that are potentially involved in plant defense against insect predation. The remaining four upregulated genes included serine peptidases and one homolog to the Arabidopsis AAA+ chaperone subunit ClpD, which is a member of the Clp protease system. Among the downregulated genes, five were homologous to serine peptidases and four were homologous to Arabidopsis Clp subunits (three homologous to Clp AAA+ chaperones and one to a ClpP-related ClpR subunit). Although the roles of serine peptidase inhibitors in plant defenses against herbivory have been extensively investigated, the roles of plant serine peptidases and the Clp protease system represent a new and underexplored field of study. The up- and downregulated D. saccharalis genes presented in this study may be candidate genes for the further investigation of the sugarcane response to herbivory.     

Protein Kinases and Parkinson’s Disease

Currently, the lack of new drug candidates for the treatment of major neurological disorders such as Parkinson’s disease has intensified the search for drugs that can be repurposed or repositioned for such treatment. Typically, the search focuses on drugs that have been approved and are used clinically for other indications. Kinase inhibitors represent a family of popular molecules for the treatment and prevention of various cancers, and have emerged as strong candidates for such repurposing because numerous serine/threonine and tyrosine kinases have been implicated in the pathobiology of Parkinson’s disease. This review focuses on various kinase-dependent pathways associated with the expression of Parkinson’s disease pathology, and evaluates how inhibitors of these pathways might play a major role as effective therapeutic molecules.     

Antioxidant and antihypertensive hydrolysates obtained from by-products of cannery sardine and brewing industries

Hydrolysates with antioxidant and antihypertensive activities were obtained from sarcoplasmic proteins of canned sardine by-product and proteases extracted from Brewer’s spent yeast. Using response surface methodology, hydrolysis time and temperature were selected to achieve the maximum bioactivity. Hydrolysates produced using the substrate/enzyme ratio 1:0.27 (mg/U), 7 h and 50ºC, presenting an angiotensin I-converting enzyme-inhibitory activity of 164 µg protein/mL and an antioxidant activity of 293 μM TE/mL. Experimental results agreed with predicted values within a 95% confidence interval. Within this work the simultaneous valorisation of two agro-industrial by-products was successfully achieved.     

Influence of frozen storage on aptitude of sardine and dolphinfish for cold-smoking process

Stability of dolphinfish (Coryphaena hippurus) and sardines (Sardina pilchardus) of different fat content (lean and fatty sardines) during frozen storage and its suitability for cold-smoking throughout storage were evaluated in order to overcome seasonal and excess catches of these species. Dolphinfish showed a relative stability regarding protein functionality (protein solubility, apparent viscosity, water and lipid holding capacity), lipid oxidation and total volatile basic nitrogen (TVBN) accumulation, which led to high acceptability ratings of the resulting smoked product throughout frozen storage (340 days). However, both lean and fatty sardines showed a marked loss of protein functionality, which coincided with the accumulation of oxidation products and TVBN. Freezing of raw muscle may become a valuable preservation method for the smoking industry to overcome the short caught period of dolphinfish in the Balearic Islands and to make use of excess catches of both, lean and fatty sardine. High quality smoked products may be obtained from the frozen muscle during approximately twelve, four and two months of frozen storage for Dolphinfish, lean and fatty sardine, respectively.     

The sequence and X-ray structure of the trypsin from Fusarium oxysporum

The trypsin from Fusarium oxysporum is equally homologous totrypsins from Streptomyces griseus, Streptomyces erythraeusand to bovine trypsin. A DFP (diisopropylfluorophosphate) inhibitedform of the enzyme has been crystallized from 1.4 M Na₂SO₄,buffered with citrate at pH 5.0–5.5. The crystals belongto space group P2₁ with cell parameters a=33.43 Å, b=67.65Å, c=39.85 Å and ß=107.6°. There isone protein molecule in the asymmetric unit. X-ray diffractiondata to a resolution of 1.8 Å were collected on film usingsynchrotron radiation. The structure was solved by molecularreplacement using models of bovine and S.griseus trypsins andrefined to an R-factor of 0.141. The overall fold is similarto other trypsins, with some insertions and deletions. Thereis no evidence of the divalent cation binding sites seen inother trypsins. The covalently bound inhibitor molecule is clearlyvisible.