An indigenous Bacillus thuringiensis strain B.t.LDC-391 producing cytocidal proteins against human colon cancer cell line, HCT-116,
was subjected to phenotypic and genotypic characterization to evaluate its relatedness to B.anthracis. The morphological features of
this strain were meta-analyzed with data of other parasporin and insecticidal protein producing Bacillus thuringiensis strains. The
conventional biochemical analysis and antibiotic sensitivity test proved it as an ampicillin resistant which is a salient feature,
absent in B.anthracis Ames. PCR analysis showed the absence of cyt and parasporin related genes in the genome of B.t.LDC-391. But
the strain was positive for cap gene. The sequencing and bio-informatic analysis of cap gene and 16S rDNA of B.t.LDC-391 placed
it closer to B.thuringiensis and revealed significant divergence from that of any B.anthracis strain. However our strain lacked β–
hemolysis on human erythrocytes which is a common feature of B.anthracis strains and parasporin producers.
Polyhydroxyalkanoates (PHAs) are natural polyesters produced by microbes, a potential alternative to synthetic plastics. Various methods ranging from gravimetry to spectrophotometry are routinely used for qualitative analysis of extracted PHA. There is a great need for accurate quantification of intracellular PHA during bioprocess. Hence, the present study aims to improvise the existing Nile red-based flow cytometry protocol. It was achieved using respective cells in a non-PHA accumulating state as gating control to minimize non-specific staining. The optimal Nile red concentration required for PHA staining is 5 × 10(3) pg mL(-1), which is ~10(3)-fold less than that of earlier reports. Further, it was inferred that flow-based quantification was more accurate than the gravimetric method. The intracellular PHA content was highest in Pseudomonas sp. MNNG-S (52.06 %) among the Pseudomonas strains tested by the flow-based method. Both gravimetric and flow-based cell cycle analyses revealed that DNA synthesis (S phase) and PHA production (log phase) are synchronous at 24-48 h of culture. This study supports flow-based PHA quantification for real time online measurement of intracellular PHA for bioreactor monitoring, control and optimization enduing industrial applications.
Cowpea (Vigna unguiculata L.) leaf discs incubated in the dark in CaCl2 and benzyladenine maintained higher levels of chlorophyll and protein than controls. The CaCl2 or benzyladenine treatments reduced lipoxygenase activity, and the effect of these compounds in combination was additive. HPLC analysis of the product profile of lipoxygenase activity with arachidonic acid as a substrate showed a single peak comigrating with standard 15‐hydroperoxyeicosatetraenoic acid. There appears to be a strong temporal correlation between the CaCl2+ benzyladenine delay of senescence and lipoxygenase activity.
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