Enantioselective Ethylation of Various Aldehydes Catalyzed by Readily Accessible Chiral Diols

Four chiral C₂‐symmetric diols were synthesized in a straightforward three‐step reaction and demonstrated excellent enantioselectivities and good overall yields. Their catalytic activities were examined via the addition of diethylzinc to various aldehydes. The enantioselective addition of diethylzinc to 2‐methoxybenzaldehyde gave the corresponding chiral secondary alcohol with high yields (up to 95%) and moderate to good enantiomeric excess (up to 88%). All synthesized ligands were evaluated in the addition of diethylzinc to various aldehydes in the presence of an additional metal such as Ti(IV) complexes. Chirality 28:593–598, 2016. © 2016 Wiley Periodicals, Inc.     

Structure-Activity Relationship and Mode of Action of a Frog Secreted Antibacterial Peptide B1CTcu5 Using Synthetically and Modularly Modified or Deleted (SMMD) Peptides

All life forms are equipped with rapidly acting, evolutionally conserved components of an innate immune defense system that consists of a group of unique and diverse molecules known as host defense peptides (HDPs). A Systematic and Modular Modification and Deletion (SMMD) approach was followed to analyse the structural requirement of B1CTcu5, a brevinin antibacterial peptide amide identified from the skin secretion of frog Clinotarsus curtipes, India, to show antibacterial activity and to explore the active core region. Seventeen SMMD-B1CTcu5 analogs were designed and synthesised by C and N-terminal amino acid substitution or deletion. Enhancement in cationicity by N-terminal Lys/Arg substitution or hydrophobicity by Trp substitution produced no drastic change in bactericidal nature against selected bacterial strains except S. aureus. But the sequential removal of N-terminal amino acids had a negative effect on bactericidal potency. Analog B1CTcu5-LIAG obtained by the removal of four N-terminal amino acids displayed bactericidal effect comparable to, or in excess of, the parent peptide with reduced hemolytic character. Its higher activity was well correlated with the improved inner membrane permeabilisation capacity. This region may act as the active core of B1CTcu5. Presence of C-terminal disulphide bond was not a necessary condition to display antibacterial activity but helped to promote hemolytic nature. Removal of the C-terminal rana box region drastically reduced antibacterial and hemolytic activity of the peptide, showing that this region is important for membrane targeting. The bactericidal potency of the D-peptide (DB1CTcu5) helped to rule out the stereospecific interaction with the bacterial membrane. Our data suggests that both the C and N-terminal regions are necessary for bactericidal activity, even though the active core region is located near the N-terminal of B1CTcu5. A judicious modification at the N-terminal region may produce a short SMMD analog with enhanced bactericidal activity and low toxicity against eukaryotic cells.     

First report of Perkinsus beihaiensis in Crassostrea madrasensis from the Indian subcontinent

Protozoan parasites of the genus Perkinsus are considered important pathogens responsible for mass mortalities in many wild and farmed bivalve populations. The present study was initiated to screen populations of the Indian edible oyster Crassostrea madrasensis, a promising candidate for aquaculture along the Indian coasts, for the presence of Perkinsus spp. The study reports the presence of P. beihaiensis for the first time in C. madrasensis populations from the Indian subcontinent and south Asia. Samples collected from the east and west coasts of India were subjected to Ray's fluid thioglycollate medium (RFTM) culture and histology which indicated the presence of Perkinsus spp. PCR screening of the tissues using specific primers amplified the product specific to the genus Perkinsus. The taxonomic affinities of the parasites were determined by sequencing both internal transcribed spacer (ITS) and actin genes followed by basic local alignment search tool (BLAST) analysis. Analysis based on the ITS sequences showed 98 to 100% identity to Perkinsus spp. (P. beihaiensis and Brazilian Perkinsus sp.). The pairwise genetic distance values and phylogenetic analysis confirmed that 2 of the present samples belonged to the P. beihaiensis clade while the other 4 showed close affinities with the Brazilian Perkinsus sp. clade. The genetic divergence data, close affinity with the Brazilian Perkinsus sp., and co-existence with P. beihaiensis in the same host species in the same habitat show that the remaining 4 samples exhibit some degree of variation from P. beihaiensis. As expected, the sequencing of actin genes did not show any divergence among the samples studied. They probably could be intraspecific variants of P. beihaiensis having a separate lineage in the process of evolution.     

Studies on the Mode of Membrane Interaction of C-terminally Amidated Brevinin1 HYba1 and 2 Peptides Against Bacteria

In this study, we investigated the possible mode of action of two C-terminally amidated novel peptides identified from the frog skin on both Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Vibrio cholerae). The results indicate that the peptides initially induce membrane depolarization followed by pore formation in a concentration-dependent manner. A microscopic examination revealed the fact that the peptides are capable of destroying bacterial cells physically. The activity of these peptides against Gram-negative bacteria was dependent on the presence of divalent cations (Ca²⁺ and Mg²⁺) but not in Gram-positive bacteria. This study also emphasizes that the endemic frogs of the Western Ghats may provide a valuable source of its skin peptides having the potential for further improvement as lead drug molecules.     

Combined evidence indicates that <Emphasis Type="Italic">Perna indica</Emphasis> Kuriakose and Nair 1976 is <Emphasis Type="Italic">Perna perna</Emphasis> (Linnaeus, 1758) from the Oman region introduced into southern India more than 100 years ago

The specific status of the brown mussel, Perna indica, in southern India and northern Sri Lanka has long been questioned. Its limited geographical distribution within the extensive range of the Asian green mussel, P. viridis, and its morphometric similarity to the African brown mussel, P. perna, have led several authorities to suggest that P. indica is not a distinct species but is in fact introduced P. perna. Analysis of DNA sequences for nuclear ITS and mitochondrial COI sequence data from newly collected mussels from southern India examined in the context of data from GenBank for mussels of the genus Perna reveals that P. indica is indeed P. perna from the Oman region. A literature review indicates that P. indica (now P. perna) has been established in southern India for at least 100 years. Reports of ecological interactions between the native P. viridis and the introduced P. perna reveal that P. perna grows to a larger size than P. viridis and outcompetes the native mussel. P. perna forms high density populations that have long been fished by local coastal communities and recently the fishery for P. perna has expanded to markets beyond the coastal area. Eradication of the introduced P. perna is not feasible and is not desirable given its important role as a source of protein and revenue for the local community. Because no monitoring of the range of P. perna in southern India occurs it is not possible to know if this species is increasing its area of distribution, but a qualitative assessment of ongoing range expansion is made based on limited evidence from published reports. The possibility exists for inter-specific hybridisation between P. viridis and P. perna and we recommend that testing commence to check for this. This research highlights the value of a detailed understanding of the genetic structure (four different clades) of the genus Perna, without which the status of P. indica as P. perna from Oman in southern India could not have been made.     

In situ pH management for microbial culture in shake flasks and its application to increase plasmid yield

Shake flasks are widely used to culture microorganisms, but they do not allow for pH control without additional infrastructure. In the presence of a carbon source like glucose, culture pH typically decreases due to overflow metabolism and can limit the growth of microorganisms in shake flasks. In this study, we demonstrate the use of magnesium hydroxide-loaded pH managing hydrogels (m-pHmH) for in situ base release to counter the decrease in culture pH in shake flasks using Escherichia coli as a model organism, in both complex and mineral salts medium. Base release from m-pHmH is shown to increase with decreasing pH (22-fold increase in release rate from pH 8 to 5), thus providing feedback from culture pH. The addition of m-pHmH resulted in better pH maintenance and higher biomass yields of E. coli K12 in media containing glucose as a carbon source. The use of m-pHmH with additional buffer resulted in pH being maintained above 6.9 while pH decreases below 5 without m-pHmH. We demonstrate one application of such in situ pH management to increase the volumetric plasmid yield from E. coli in shake flask culture. In situ glucose release through a hydrogel to mimic fed-batch culture along with the addition of m-pHmH resulted in a 395 % increase in volumetric plasmid yield to 38 μg/ml in shake flask culture.