Characterization and biomedical application of Tetradecamethylcycloheptasiloxane, a silicone-type biosurfactant produced by Streptomyces castaneoglobisporus AJ9 isolated from solar salt works

694-704Tetradecamethylcycloheptasiloxane (C14H42O7Si7), a biosurfactant, was extracted by acid precipitation method from the haloalkaliphilic actinomycetes, Streptomyces castaneoglobisporus AJ9 (Gene Bank KC603900.1) and characterized by GC-MS analysis. The purified biosurfactant could effectively degrade dyes such as orange MR, direct violet, cotton red, reactive yellow and nitro green. Antimicrobial screening results showed that biosurfactants could effectively control bacterial pathogens such as Staphylococcus aureus and Vibrio parahaemolyticus and fungal pathogens such as Fusarium sp. and Penicillium sp. Even a small diminutive quantity of biosurfactant (100 μg) could totally block the multiplication of White Spot Syndrome Virus (WSSV) in shrimp in vivo. The suppression of cancer cells in vitro by the biosurfactant was 74 % as confirmed by Response Surface Methodology (RSM) and residual plot analysis. Wheat bran, groundnut oilcake and oilseed cake seem to enhance biosurfactant production

cDNA cloning and expression of vitamin-K Epoxide Reductase (VKOR) gene from orange spotted grouper <i>Epinephelus coioides</i><i><span style="font-family:"Book Antiqua"; mso-bidi-font-family:Arial" lang="EN-US"></span></i>

573-579The cDNA sequence encoding Vitamin-K Epoxide Reductase (VKOR) was constructed from hypothalamus and pituitary cDNA libraries of Orange spotted Grouper Epinephelus coioides. VKOR gene contains 610 nucleotides including the open reading frame (ORF) of length 399 bp.  ORF was started with ATG codon at the position of 199 and terminated with a stop codon TAA of 585.  ORF sequence had 128 amino acids and the predicted protein molecular weight is approximately 14.08 kDa (GenBank: FJ939281.1). Expression of VKOR gene in different tissues was further examined using one-step RT-PCR. RT PCR result revealed that, VKOR mRNA expression was detected only in liver, kidney, hypothalamus and pituitary. VKOR gene sequence of E. coioides aligned with related sequences by multiple sequence alignment by clustal X and phylogenetic tree was generated by Neighbor Joining method using Genious v 5.4. E. coioides VKOR had the highest identity and homology with the puffer fish, Tetraodon nigroviridis, Takifugu rubripes and Zebra fish Danio rerio. Based on the VKOR expression in the kidney, liver hypothalamus and pituitary of the seven year’s old (sex reversal stage) E. coioides, it may be involved in sex reverse

<em>Streptomyces castaneoglobisporus</em> AJ9, A haloalkaliphilic actinomycetes isolated from solar salt works in southern India and its pharmacological properties

475-488Five strains of haloalkaliphilic Streptomyces sp. were isolated and identified. Among the five strains, the Streptomyces sp. AJ9 which isolated Kovalam solar salt works had the potential ability of protease, gelatinase enzymes production and highly antagonistic against the pathogenic bacteria including Aeromonas hydrophila, Escherichia coli, Salmonella typhi, Staphylococcus aureus and Vibrio parahaemolyticus. The Streptomyces sp. AJ9 was identified by genomic level through 16S rRNA sequencing and confirmed as Streptomyces castaneoglobisporus AJ9 (GenBank KC603900.1). Further the secondary metabolites were extracted with ethyl acetate, purified through silica column chromatography and screened the fractions (FI to FVII) against pathogenic bacteria, fungi, the shrimp killer virus, White Spot Syndrome Virus (WSSV). Among the fractions FIV to FVI had highest bioactivities and the pooled fractions were structurally characterized by GC-MS analysis and confirmed as the benzoic acid derivatives of 2-hydroxy-4-methoxy-6-methyl-, methyl ester. The pooled fraction (FIV to FVI) were significantly (Pin vitro levels and able to kill the L 929 cancer cells at the rate of 75.58 % µg/ml

cDNA cloning and expression of vitamin-K Epoxide Reductase (VKOR) gene from orange spotted grouper Epinephelus coioides

The cDNA sequence encoding Vitamin-K Epoxide Reductase (VKOR) was constructed from hypothalamus and pituitary cDNA libraries of Orange spotted Grouper Epinephelus coioides. VKOR gene contains 610 nucleotides including the open reading frame (ORE) of length 399 bp. ORE was started with ATG codon at the position of 199 and terminated with a stop codon TAA of 585. ORE sequence had 128 amino acids and the predicted protein molecular weight is approximately 14.08 kDa (GenBank: FJ939281.1). Expression of VKOR gene in different tissues was further examined using one-step RT-PCR. RT PCR result revealed that, VKOR mRNA expression was detected only in liver, kidney, hypothalamus and pituitary. VKOR gene sequence of E. coioides aligned with related sequences by multiple sequence alignment by clustal X and phylogenetic tree was generated by Neighbor Joining method using Genious v 5.4. E. coioides VKOR had the highest identity and homology with the puffer fish, Tetraodon nigroviridis, Takifugu rubripes and Zebra fish Danio rerio. Based on the VKOR expression in the kidney, liver hypothalamus and pituitary of the seven year's old (sex reversal stage) E. coioides, it may be involved in sex reverse.The cDNA sequence encoding Vitamin-K Epoxide Reductase (VKOR) was constructed from hypothalamus and pituitary cDNA libraries of Orange spotted Grouper Epinephelus coioides. VKOR gene contains 610 nucleotides including the open reading frame (ORE) of length 399 bp. ORE was started with ATG codon at the position of 199 and terminated with a stop codon TAA of 585. ORE sequence had 128 amino acids and the predicted protein molecular weight is approximately 14.08 kDa (GenBank: FJ939281.1). Expression of VKOR gene in different tissues was further examined using one-step RT-PCR. RT PCR result revealed that, VKOR mRNA expression was detected only in liver, kidney, hypothalamus and pituitary. VKOR gene sequence of E. coioides aligned with related sequences by multiple sequence alignment by clustal X and phylogenetic tree was generated by Neighbor Joining method using Genious v 5.4. E. coioides VKOR had the highest identity and homology with the puffer fish, Tetraodon nigroviridis, Takifugu rubripes and Zebra fish Danio rerio. Based on the VKOR expression in the kidney, liver hypothalamus and pituitary of the seven year's old (sex reversal stage) E. coioides, it may be involved in sex reverse