Heterologous expression, purification, and phylogenetic analysis of oil-degrading biosurfactant biosynthesis genes from the marine sponge-associated Bacillus licheniformis NIOT-06.

Surfactin is a lipopeptide, composed of one ß-hydroxy fatty acid, a long fatty acid moiety, and seven amino acids. In this study, the biosurfactant biosynthesis genes; 4'-pantetheinyl transferase (sfp), phosphopantetheinyl transferase (sfpO), and surfactin synthetase (srfA) have been characterized from the marine sponge-associated Bacillus licheniformis NIOT-06 from the Andaman and Nicobar Islands. The purified recombinant biosurfactant revealed excellent emulsification activity with crude oil and kerosene. Reverse-phase high-performance liquid chromatography resolved the purified recombinant biosurfactant into several fractions and one of which had significant surface tension reducing property. Fourier transform infrared spectroscopy spectrum also revealed the presence of C-N-N, alkenes, and N-H as the functional groups, and a similar overlapping pattern was observed with that of standard lipopeptide surfactin. The diversity and phylogeny of sfp, sfpO, and srfA gene sequences were compared with other eubacteria. The sfp, sfpO, and srfA gene sequences obtained from Bacillus licheniformis NIOT-06 were diverse and appeared to be partially conserved when compared with the GenBank reported sequences of several eubacteria.

Complex bacterial communities in the deep-sea sediments of the Bay of Bengal and volcanic Barren Island in the Andaman Sea.

Deep-sea environments are gaining global attention as potential sources of useful microorganisms, thereby warranting a better understanding of the diversity and genomic potential of the microbes present. To this end, here we provide the first insights into the composition of the bacterial communities in deep-sea sediment samples from the southwestern Bay of Bengal and the geographically distinct volcanic Barren Island in the Andaman Sea. High-throughput 16S rRNA gene sequencing of the sediments revealed the presence of >44,000 operational taxonomic units (OTUs) in each of the samples, suggesting high bacterial diversity. Actinobacteria was the most dominant phylum, representing >20% of the taxonomically assignable OTUs, followed by Firmicutes, Proteobacteria, and Chloroflexi. Numerous bacteria that are potentially involved in the sulfur cycle were observed in the Barren Island sediment sample, while bacteria with clinical and industrial potential were observed in the samples from the southwestern Bay of Bengal. Correlation analysis of the biotic and abiotic parameters showed that the differences in bacterial richness and community composition between the sampling sites were mainly dependent on sediment texture. Using a predictive functional metagenomic approach, this study also discusses the genetic variations that may provide an adaptive advantage to sediment bacterial communities for survival in these extreme deep-sea environments. The results from this study should aid future studies focused on bioprospecting and geochemical cycling in the deep sea.

Molecular expression of l-asparaginase gene from Nocardiopsis alba NIOT-VKMA08 in Escherichia coli: A prospective recombinant enzyme for leukaemia chemotherapy.

l-Asparaginase is an antineoplastic agent that selectively reduces the level of l-asparagine in blood and diminishes the proliferation of cancerous cells. Studies were carried out on the cloning and heterologous expression of l-asparaginase biosynthesis gene (ansA) from Nocardiopsis alba NIOT-VKMA08 to achieve the stable inducible system that overproduces the glutaminase-free recombinant l-asparaginase. Overexpression of recombinant l-asparaginase was achieved with an optimized final concentration of 1.5mM of isopropyl-ß-d-thiogalactoside (IPTG) and the enzyme was expressed as a soluble protein. The recombinant enzyme was purified using nickel-nitrilotriacetic acid (Ni-NTA) chromatography and the purified enzyme disclosed an elevated level of asparaginase activity (158.1IU/mL). Optimum pH and temperature of the purified l-asparaginase for the hydrolysis of l-asparagine were 8.0 and 37°C and it was very specific for its natural substrate, l-asparagine. Detailed studies were carried out on the kinetics of enzyme reaction, catalytic activity, temperature and ionic strength and the thermostability of the l-asparaginase enzyme. The functional characterisation of the recombinant l-asparaginase was studied through Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), in silico sequence analysis and protein structural modelling. Glutaminase activity was not detected in the recombinant l-asparaginase, which could reduce the probable side effects during leukaemia therapy.

Enterococcus species diversity and molecular characterization of biomarker genes in Enterococcus faecalis in Port Blair Bay, Andaman and Nicobar Islands, India.

This study was performed to evaluate the abundance and diversity of Enterococcus sp. and the distribution of biomarker genes in Enterococcus faecalis in Port Blair Bay, Andaman and Nicobar Islands. The Enterococcus sp. densities at the seven sampling stations were highly influenced by tidal fluctuations and season. The distributions and diversities of species varied in the inner and outer regions of Port Blair Bay. Among the 1816 total isolates, the occurrence of fecal Enterococcus was high (1.78×10(4) CFU/100 mL) in Phoenix Bay. Moreover, 67.76% of the isolates were identified as Enterococcus, and the most frequently identified species were E. hirae, E. avium and E. faecalis. Assessments of antibiotic resistance and biomarker genes revealed the maximum occurrence in the Aberdeen Bay isolates. The most prevalent biomarker genes observed in the E. faecalis isolates were gelE and asa1, whereas cyl was not found among the isolates. In silico sequence analysis of biomarker genes of E. faecalis also revealed that they are evolutionarily well conserved with those of earlier reports. Further, multivariate analysis distinguished the JB, PB and OS stations from the other stations according to distinctive microbial densities and compositions. In addition, the Shannon-Wiener diversity indices and box-whisker plots further facilitated and supported the multivariate results.