Polyhydroxybutyrate production from marine source and its application.

The increasing significance of non-degradable plastic wastes is an emerging concern. As a substitute, researches are being endeavoured from existing reserve to yield bioplastics based on their properties of biodegradability. Owing to their cost, now the experts are quest for a substitute source like bacteria, microalgae, actinomycetes, cyanobacteria and plants. PHB is biodegradable, environmental friendly and biocompatible thermoplastics. Varying in toughness and flexibility, depending on their formulation, they are used in various ways similar to many non-biodegradable petrochemical plastics currently in use. Promising strategies contain genetic engineering of microorganisms to introduce production pathways examined for the past two decades. Such kind of researches focusing on the use of unconventional substrates, novel extraction methods, and genetically enhanced species with assessment to make PHB from marine microbes are commercially attractive field. Hence, this biopolymer synthesis may displayed as one of the survival mechanisms of endosymbiotic, macroalgae, or sponge-associated bacteria, which exist in a highly competitive and stressful marine microenvironment. This review throws light on the promising and growing awareness of using marine microbes as PHB source, along with their applications in different fields of aquaculture, medicine, antifouling and tissue engineering.

Synthesis of chitosan capped copper oxide nanoleaves using high intensity (30kHz) ultrasound sonication and their application in antifouling coatings.

The synthesis of chitosan capped copper oxide nanoleaves (CCCO NLs) was carried out under three different reaction conditions viz. 1) room temperature, 2) 70°C and 3) high intensity ultrasound (30kHz) sonication method and it has been found that the high intensity ultrasound (30kHz) sonication is the best method when compared to other two methods. The advantages of the present synthetic method are: i) easy one step process, ii) lesser reaction time, iii) good yield, iv) reproducible and v) calcination is not required. The resulting chitosan capped copper oxide nanoleaves were characterized by Diffuse Reflectance UV-Visible Spectroscopy (DRS), Fourier Transform Infra-Red Spectroscopy (FT-IR), X-ray Diffraction (XRD), X-ray Photoelectron Spectroscopy (XPS), Field Emission Scanning Electron Microscopy (FESEM), High Resolution Transmission Electron Microscopy (HRTEM) and Thermo gravimetric analysis (TGA). The CCCO NLs were blended with commercial paints such as polyurethane clear, polyurethane white and acrylic emulsion and applied on to three different surfaces (wood, mild steel and cement slab panels). The hydrophilicity of CCCONP coated panels was analyzed by water contact angle measurement and their antifouling behavior was investigated against three different green and marine algae viz. Arthrospira, Chlorella and Amphora. The antifouling efficiency of the CCCO NLs against the algae was found to be 78-92%.

Mass cultivation of UV-B adapted Arthrospira platensis RRGK under open raceway pond for the production of Poly-ß-hydroxy butyrate.

Six different strains of cyanobacteria were isolated from the freshwater lake, Arakkonam, India. Staining of cells with Nile Red showed the presence of large quantities of PHB granules in the cell cytoplasm of Arthrospira. Molecular identification of the strain was carried out using 16S rRNA analysis and their systematic position was ascertained as Arthrospira platensis RRGK. Studies were carried out on random mutagenesis approach using UV-B radiation for enhancing the production of PHB. Further, Response Surface Methodology was used for optimization of pH, temperature, and sodium bicarbonate for higher biomass and PHB production. Under open raceway pond A. platensis RRGK produced biomass concentration of 2.2±0.13gL-1 and 131±0.36gL-1 of PHB content. It was chemically characterized through FTIR, DSC, TGA and XRD analyses. Hence, PHB can be produced from cyanobacteria by sequestering harmful CO2. It can also be used as a substitute for synthetic polymers in tissue engineering.

Optimization of polyhydroxybutyrate production utilizing waste water as nutrient source by Botryococcus braunii Kütz using response surface methodology.

Investigations have been made to optimize various factors including pH, temperature, and substrate for enhanced polyhydroxybutyrate (PHB) production in Botryococcus braunii which serves as a pioneer for production of bioplastic (PHB). Polyhydroxybutyrate is a natural, decomposable polymers accumulated by the microorganism under different nutritional condition. Strain selection was done by staining method using Sudan black and Nile red dye. Using response surface methodology (RSM), three level- three variables Box Behnken design (BBD), the best potential combination of pH (4-11), temperature (30-50°C) and sewage waste water as substrate fed at different concentrations at 20%-100% for maximum PHB production was investigated. Maximum yield (247±0.42mg/L) of PHB dry weight was achieved from the 60% concentration of sewage waste water as a growth medium at pH 7.5 at 40°C. It was well in close agreement with the value predicted by RSM model yield (246± 0.32mg/L). Thus the study shows the production of PHB by B. braunii along with the basic characterization of PHB by using FTIR and TEM analysis. These preliminary studies indicated that PHB can also be produced by B. braunii utilizing waste water. There is no report on the optimization of PHB production in this microalgae have been documented.

Biosynthesis, purification and characterization of polyhydroxybutyrate from Botryococcus braunii kütz.

Polyhydroxybutyrate (PHB) is completely biodegradable which is metabolised by microorganisms in the soil as their sole food source in few years. The level of PHB up to 10.6% of algal dry weight is of great potential of the eco-friendly nature. Botryococcus braunii is mainly used for the production of biodiesel and is also capable of producing biopolymer polyhydroxy butyrate (PHB). In this study, Botryococcus braunii is used which generally produce PHB to around 20% of the dry weight. Three different microalgae were isolated from the fresh water of Kolavoi lake of Tamil Nadu. They were identified by their morphological features under the light microscope. The primary screening of PHB intracellular granules was made by using Nile red dye under a fluorescent microscope. Among them, Botryococcus braunii showed high accumulation of PHB granules. For authentic confirmation, the chloroform extracted PHB was analysed by FTIR, XRD and DSC-TGA analyses to characterize PHB with commercial biodegradable thermoplastic. This is the first report in B. braunii for its PHB production.

Further studies and biological activities of macromolecular protein R-Phycoerythrin from Portieria hornemannii.

In the present study, the purified R-Phycoerythrin (R-PE) from a red alga Portieria hornemannii was subjected to the analysis of stability under the influence of different agents. Among the various inhibitors tested on R-PE EDTA at lower concentrations (<1 mM) supported the activity of R-PE. When R-PE was exposed to different organic solvents, ethanol supported the activity at the maximum followed by acetone, ethyl acetate, chloroform and methanol. Citric acid, as a preservative maintained the stability of R-PE both under 0 ± 5 °C and 30 ± 5 °C with 59.34% and 56.23% respectively, on 30th day. Thermal decomposition of the R-PE began near 60 °C. Maximum weight loss was occurred between 150 °C and 500 °C. Complete weight loss was recorded around 875 °C. Thermal denaturation was observed between 19 °C and 40 °C. Moderate to low antioxidant activities were observed in R-PE in relation to total antioxidant activities. After characterization, R-PE was taken for in vitro anticancer studies against selected cancer cell lines. Further studies involving AO/EB fluorescence staining and phase contrast microscope revealed characteristic apoptotic features like cell shrinkage, membrane blebbing, and nuclear DNA fragmentation, etc. Likewise, FACS analysis revealed the cell cycle distribution pattern of A549 and HepG2 cells.

Isolation and characterization of macromolecular protein R-Phycoerythrin from Portieria hornemannii.

R-Phycoerythrin (R-PE) is one of the three phycobiliproteins which are extensively used as fluorescent probes, and it is prepared from red macro-algae. This macromolecular protein has gained importance in many biotechnological applications in food science, immunodiagnostic, therapy, cosmetics, protein and cell labeling, and analytical processes. In the present investigation, R-PE was isolated and purified from a red alga Portieria hornemannii. R-PE extracted and purified through ammonium sulfate precipitation (55%) followed by Q-Sepharose column chromatography had yielded a maximum purity of 5.2%. R-PE exhibited a typical "three-peak" with absorption maxima at 499, 545 and 565 nm. CD spectrum of R-PE yielded the following secondary structure data: alpha helix (14.30%), beta helix (28.10%), turn helix (19.20%) and random coil helix (38.40%). The molecular mass of R-PE was 240 kDa under Native-PAGE. Three different subunits such as α, ß and γ of 16 kDa, 21 kDa and 39 kDa were segregated under SDS-PAGE. On two dimensional gel electrophoresis, one basic and four acidic subunits were detected. Five different tryptic peptides were assigned under MALDI-TOF. The sequences of N-terminus of R-PE of 10 different amino acids are Met Lys Gln Met Trp Asp Arg Met Val Val. The preparative procedures of the R-PE extraction and purification established based on the experiments exhibit advantages and can offer a reference for R-PE preparation from other marine red macro-alga P. hornemannii.