Enzyme Bioprospection of Marine-Derived Actinobacteria from the Chilean Coast and New Insight in the Mechanism of Keratin Degradation in Streptomyces sp. G11C

Marine actinobacteria are viewed as a promising source of enzymes with potential technological applications. They contribute to the turnover of complex biopolymers, such as pectin, lignocellulose, chitin, and keratin, being able to secrete a wide variety of extracellular enzymes. Among these, keratinases are a valuable alternative for recycling keratin-rich waste, which is generated in large quantities by the poultry industry. In this work, we explored the biocatalytic potential of 75 marine-derived actinobacterial strains, focusing mainly on the search for keratinases. A major part of the strains secreted industrially important enzymes, such as proteases, lipases, cellulases, amylases, and keratinases. Among these, we identified two streptomycete strains that presented great potential for recycling keratin wastes—Streptomyces sp. CHA1 and Streptomyces sp. G11C. Substrate concentration, incubation temperature, and, to a lesser extent, inoculum size were found to be important parameters that influenced the production of keratinolytic enzymes in both strains. In addition, proteomic analysis of culture broths from Streptomyces sp. G11C on turkey feathers showed a high abundance and diversity of peptidases, belonging mainly to the serine and metallo-superfamilies. Two proteases from families S08 and M06 were highly expressed. These results contributed to elucidate the mechanism of keratin degradation mediated by streptomycetes.     

Glutamic acid supplementation recovers the reduced performance of weanling pigs fed reduced crude protein diets

This study was conducted to evaluate the supplementation of glutamic acid(Glu)to reduced protein diets on the performance of weanling pigs.One hundred and eighty crossbred weanling pigs([Yorkshire
Landrace]
Duroc,21 d old)having similar body weight(BW)of 6.45 kg were randomly allotted to 1 of 6 dietary treatments(5 pigs per pen[2 barrows and 3 gilts];6 pens per treatment)based on BW and sex during a 6-week trial.Dietary treatments consisted of positive control(PC)diet formulated to have 226.9,205.6,and 188.8 g crude protein(CP)during phases 1,2,and 3,respectively,and negative control(NC)diets with 20 g CP reduction from PC diets and addition of Glu with increasing levels,resulting in the calculated Lys-to-Glu ratios of 1:2.25,1:2.30.1:2.35,1:2.40,and 1:2.45,designated as NC,NC1,NC2,NC3,and NC4,respectively.The BW of pigs receiving PC diet was higher(P<0.05)than those receiving NC diet at d 7,21 and 42.A higher(P<0.05)average daily gain(ADG)from d 1 to 7,8 to 21,22 to 42 and during the overall experiment period was observed in pigs fed PC than NC diet.Pigs fed NC diets including the graded level of Glu linearly increased(P<0.05)BW at d 42,ADG and gain-to-feed ratio(G:F)during the overall experimental period.In addition,trends in linear increase in BW(P=0.056)at d 7 and ADG from d 1 to 7 and d 22 to 42(linear effect,P=0.081,P=0.058 respectively)were observed.A tendency in the linear increment of NH3(P=0.082)at d 21 and linear reduction in methyl mercaptans(P=0.054)emission at d 42 was observed in pigs fed NC diets supplemented with graded level of Glu.In conclusion,supplementing the reduced protein diet with Glu enhanced the growth performance in weanling pigs suggesting that supplementation of Glu can compensate the reduction of 2%CP in the basal diets.     

Viewpoints on synergising ASEAN academic visibilities through research collaboration and the establishment of an ASEAN Citation Index Database1

This opinion article expresses two key viewpoints regarding the options for Association of South‐East Asian Nations (ASEAN) universities in pursuit of fulfilling the demand for greater research visibility and academic reputation. The first viewpoint outlines the importance of promoting research collaboration among ASEAN researchers and their international peers in increasing the visibility and impact of ASEAN research. The second viewpoint examines the ASEAN research characteristics, the challenges and existing best practices of its journal publishing landscape, the potential role of non‐ASEAN international journals in improving the profile of ASEAN research and explores the needs and benefits of establishing an ASEAN Integrated Journal Publishing Network (IJPN). It is through these combined approaches, we believe, that effective policy change, the standardisation of publishing structure, procedures, and systems to drive journal development, improvement of journal quality, and establishment of a consistent framework for measuring researcher performance will result. This article also includes a proposal for the IJPN structure and implementation plan, and highlights the key benefits of taking a collaborative approach to achieving this common goal.     

Parental participation in progeny and effective population sizes in experimental seed orchards of wild cherry Prunus avium L. (Batsch)

Parents and progenies genetic diversity, and male and female contributions to the seed crop were assessed in three experimental Prunus avium seed orchards. Collected data were used to compare different effective population sizes, based on phenological, seed crop and paternity analysis. Our results did not show any difference of genetic diversity between parents and progenies. A limited pollen pollution was detected. We showed that distance and coflowering among clones had a significant effect on effective pollination, and a significant effect of the production of flowers was revealed in one of the seed orchards. Our study also revealed a quite low number of effective size of fathers per mother, but high effective sizes of mothers, fathers and parents at the level of the seed orchard. Finally, the calculation of effective size of mothers, fathers and parents was not highly modified when having the complete information based on the paternity analysis.     

Macrostachyols A-D, new oligostilbenoids from the roots of Gnetum macrostachyum

Four new oligostilbenes, macrostachyols A-D (1-4) along with six known stilbenoids (5-10), were isolated from the roots of Gnetum macrostachyum. These compounds were also evaluated for their cytotoxicity towards HeLa and KB cell lines. Only compound 4 showed the significant cytotoxicity to HeLa cells with IC₅₀ value of 4.13 μM.     

A new dimeric aporphine from the roots of Artabotrys spinosus

A new dimeric aporphine, artabotrysine along with five known compounds was isolated from the roots of Artabotrys spinosus. Their structures were fully established on the basis of spectral evidence. All isolated compounds were evaluated for their cytotoxicity on HeLa and KB cells.     

Atomic structure of thymidylate synthase: target for rational drug design

The atomic structure of thymidylate synthase from Lactobacillus casei was determined at 3 angstrom resolution. The native enzyme is a dimer of identical subunits. The dimer interface is formed by an unusual association between five-stranded beta sheets present in each monomer. Comparison of known sequences with the Lactobacillus casei structure suggests that they all have a common core structure around which loops are inserted or deleted in different sequences. Residues from both subunits contribute to each active site. Two arginine side chains can contribute to binding phosphate on the substrate. The side chains of several conserved amino acids can account for other determinants of substrate binding.     

Stable amplified DNA in drug-resistant Leishmania exists as extrachromosomal circles

The relative stability of amplified DNA in drug-resistant Leishmania major was previously reported to be dependent on location, that is, unstable amplified DNA was extrachromosomal and stable amplified DNA was chromosomal. Leishmanial chromosomes have now been directly examined by means of orthogonal-field-alternation gel electrophoresis (OFAGE). The amplified DNA's in three resistant cell lines displayed unusual migration and were clearly extrachromosomal, regardless of whether the amplified DNA's were stable or unstable. Thus, contrary to conclusions from earlier studies of drug resistance in cultured animal cells, stable amplified DNA in Leishmania can be extrachromosomal. In addition, these amplified DNA's were shown to be circular on the basis of their resistance to exonuclease III digestion and their behavior on OFAGE. Their mobility was also greatly changed after treatment with topoisomerase II, suggesting that the amplified DNA's were either supercoiled or concatenated circles.     

The stress of weaning influences serum levels of acute-phase proteins, iron-binding proteins, inflammatory cytokines, cortisol, and leukocyte subsets in Holstein calves

The purpose of our study was to investigate changes in immunological parameters induced by weaning stress (including milk restriction) in calves. Fifteen Holstein calves were subjected to weaning at 6 weeks of age. Blood samples were collected at -14, -7, -2, 1, 3, and 5 days post-weaning (DPW; 0 DPW = 42 days). Weaning caused significant (p < 0.01) increases in the neutrophil (NE):lymphocyte (LY) ratio at 5 DPW with a significant (p < 0.05) reduction of LYs. The concentration of acute-phase proteins (haptoglobin and serum amyloid A) also increased significantly (p < 0.05) at 3 and 5 DPW compared to -2 DPW. Levels of the iron-binding protein lactoferrin decreased significantly (p < 0.05) after weaning. Serum tumor necrosis factor-α and cortisol levels were elevated (p < 0.05) at 3 DPW, while those of serum interferon-γ decreased (p < 0.05) at 1 and 3 DPW compared to levels observed before weaning. Weaning significantly (p < 0.05) decreased the percentage of CD25⁺ T cells in the peripheral blood. In conclusion, weaning stress affected the NE:LY ratio along with the levels of acute phase proteins, lactoferrin, cortisol, and inflammatory cytokines in the peripheral blood of calves. Weaning stress may induce an acute phase response possibly through the elevation of cortisol production and modulation of inflammatory cytokines.