Shailesh S. Sawant

Microorganisms as efficient biosystem for the synthesis of metal nanoparticles: current scenario and future possibilities

Nanoparticles, the elementary structures of nanotechnology, are important materials for fundamental studies and variety of applications. The different sizes and shapes of these materials exhibit unique physical and chemical properties than their bulk materials. There is a great interest in obtaining well-dispersed, ultrafine, and uniform nanoparticles to delineate and utilize their distinct properties. Nanoparticle synthesis can be achieved through a wide range of materials utilizing a number of methods including physical, chemical, and biological processes with various precursors from liquids and solids. There is a growing need to prepare environmentally friendly nanoparticles that do not produce toxic wastes in their process synthesis protocol. This kind of synthesis can be achieved by green environment benign processes, which happen to be mostly of a biological nature. Microorganisms are one of the most attractive and simple sources for the synthesis of different types of nanoparticles. This review is an attempt to provide the up-to-date information on current status of nanoparticle synthesis by different types of microorganisms such as fungi, yeast, bacteria, cyanobacteria, actinomycete, and algae. The probable biosynthesis mechanism and conditions for size/shape control are described. Various applications of microbially synthesized nanoparticles are summarized. They include antibacterial, antifungal, anticancer, larvicidal, medical imaging, biosensor, and catalytic applications. Finally, limitations and future prospects for specific research are discussed.

Degradation of corn stover by fungal cellulase cocktail for production of polyhydroxyalkanoates by moderate halophile Paracoccus sp. LL1.

Bioprocessing of lignocellulose as a renewable resource for fuels, chemicals or value added products is a necessity to fulfil demands of petroleum products. This study aims to convert corn stover to polyhydroxyalkanoates (PHA). Corn stover was hydrolyzed to crude sugars by an on-site prepared cellulase cocktail from co-culture of Trichoderma reesei and Aspergillus niger. The potent PHA producer, Paracoccus sp. LL1, was isolated from Lonar Lake, India and could accumulate PHA up to 72.4% of its dry cell weight. PHA production reached 9.71 g/L from corn stover hydrolysate containing 40 g/L sugar mixture. The PHA synthase gene (phaC) sequence of the isolate showed 79% identity with the phaC gene of Paracoccus seriniphilus (E71) strain from the NCBI database. The nature/type of PHA was found to be poly(3-hydroxybutyrate) by Fourier transform infrared spectroscopy.

A rapid, sensitive, simple plate assay for detection of microbial alginate lyase activity

Screening of microorganisms capable of producing alginate lyase enzyme is commonly carried out by investigating their abilities to grow on alginate-containing solid media plates and occurrence of a clearance zone after flooding the plates with agents such as 10% (w/v) cetyl pyridinium chloride (CPC), which can form complexes with alginate. Although the CPC method is good, advantageous, and routinely used, the agar in the media interferes with the action of CPC, which makes judgment about clearance zones very difficult. In addition, this method takes a minimum of 30 min to obtain the zone of hydrolysis after flooding and the hydrolyzed area is not sharply discernible. An improved plate assay is reported herein for the detection of extracellular alginate lyase production by microorganisms. In this method, alginate-containing agar plates are flooded with Grams iodine instead of CPC. Grams iodine forms a bluish black complex with alginate but not with hydrolyzed alginate, giving sharp, distinct zones around the alginate lyase producing microbial colonies within 2-3 min. Grams iodine method was found to be more effective than the CPC method in terms of visualization and measurement of zone size. The alginate-lyase-activity area indicated using the Grams iodine method was found to be larger than that indicated by the CPC method. Both methods (CPC and Grams iodine) showed the largest alginate lyase activity area for Saccharophagus degradans (ATCC 43961) followed by Microbulbifer mangrovi (KCTC 23483), Bacillus cereus (KF801505) and Paracoccus sp. LL1 (KP288668) grown on minimal sea salt medium. The rate of growth and metabolite production in alginate-containing minimal sea salt liquid medium, followed trends similar to that of the zone activity areas for the four bacteria under study. These results suggested that the assay developed in this study of Grams iodine could be useful to predict the potential of microorganisms to produce alginate lyase. The method also worked well for screening and identification of alginate lyase producers and non-producers from environmental samples on common laboratory media. They did this by clearly showing the presence or absence of clearance zones around the microbial colonies grown. This new method is rapid, efficient, and could easily be performed for screening a large number of microbial cultures. This is the first report on the use of Grams iodine for the detection of alginate lyase production by microorganisms using plate assay.

Rapid biological synthesis of silver nanoparticles using Kalopanax pictus plant extract and their antimicrobial activity

Silver nanoparticles (AgNPs) have promising potential in biomedicine, energy science, optics, and health care applications. We synthesized AgNPs using plant, Kalopanax pictus leaf extract. UV-visible spectrophotometric study showed the characteristic peak for AgNPs at wavelength 430 nm. The optical density at 430 nm increased after addition of plant leaf extract, indicating increase in formation of nanoparticles. Comparative time course analyses for AgNP synthesis carried out at different reaction temperatures (20, 60, and 90 °C) revealed higher reaction rate for K. pictus than Magnolia kobus plant leaf extract, which showed highest AgNP synthesis rate in the previous report. Electron microscopy analyses confirmed the presence of well dispersed AgNPs, predominantly with spherical shapes. In transmission electron microscopy, the particle size decreased with increase in temperature. Electron dispersive X-ray spectroscopy analyses indicated that Ag content increased with increase in reaction temperature. Fourier transform-infrared spectroscopy studies revealed capping of bioorganics from plant to the synthesized AgNPs. The antimicrobial activity of the synthesized AgNPs against Escherichia coli increased with increase in reaction temperature. The observations in this study will prove beneficial in approaching rapid synthesis of AgNPs and their antimicrobial application.

Lignocellulosic and marine biomass as resource for production of polyhydroxyalkanoates

Polyhydroxyalkanoates (PHAs) are considered as sustainable ‘green/bio plastics’ because they have potential to replace their depleting petroleum-based competitors in the recent future. To reach this goal, PHAs must be able to compete with the established petroleum-based plastics in both technical and economic aspects. The current PHA production is based on high-priced substrates of high nutritional value and simple carbon sources such as glucose, sucrose, starch, or vegetable oils. Non-food based carbon-rich complex polysaccharides of lignocellulosic and marine biomass can be used as alternative and suitable feedstock through consolidated bioprocessing (CBP). CBP is a promising strategy that involves the production of lytic enzymes, hydrolysis of biomass, and fermentation of resulting sugars to desired products in a single process step. CBP offers very large cost reductions if microorganisms possessing the abilities are found or microbial processes are developed to utilize substrate and simultaneously produce products. This review focuses on possible available complex polysaccharides of lignocellulosic and marine biomass that can be used as resources to produce PHAs in biorefineries, including CBP.

Potential of biosynthesized silver nanoparticles as nanocatalyst for enhanced degradation of cellulose by cellulase

Silver nanoparticles (AgNPs) as a result of their excellent optical and electronic properties are promising catalytic materials for various applications. In this study, we demonstrate a novel approach for enhanced degradation of cellulose using biosynthesized AgNPs in an enzyme catalyzed reaction of cellulose hydrolysis by cellulase. AgNPs were synthesized through reduction of silver nitrate by extracts of five medicinal plants (Mentha arvensis var. piperascens, Buddleja officinalisMaximowicz, Epimedium koreanum Nakai, Artemisia messer-schmidtiana Besser, and Magnolia kobus). An increase of around twofold in reducing sugar formation confirmed the catalytic activity of AgNPs as nanocatalyst. The present study suggests that immobilization of the enzyme onto the surface of the AgNPs can be useful strategy for enhanced degradation of cellulose, which can be utilized for diverse industrial applications.

Production of polyhydroxyalkanoates by Ralstonia eutropha from volatile fatty acids

Polyhydroxyalkanoates (PHAs) are biodegradable and biocompatible thermoplastics that can be synthesized in various microorganisms. Volatile fatty acids (VFAs) are produced by anaerobic treatment of organic wastes that can be utilized as inexpensive substrates for PHA synthesis. In this study, several Ralstonia eutropha strains were grown on the mixture of VFAs (acetic, propionic, and butyric acid) as its carbon and energy source for growth and PHA synthesis. R. eutropha KCTC 2658 accumulated PHAs up to 50% of dry cell weight from total 5 g/L of mixed VFAs (acetic acid: propionic acid: butyric acid=1: 2: 2). In batch culture of R. eutropha KCTC2658 in a 5 L fermentor, a homopolymer of poly(3-hydroxybutyrate) [P(3HB)] was produced from 20 g/L glucose as a sole carbon source with dry cell weight of 8.4 g/L and PHA content of 30%. In fed-batch culture, two feeding strategies, pulse or pH-stat, were applied to add VFAs to the fermentor. When VFAs were fed using pH-stat feeding strategy after 40 h, a copolymer of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] was produced with dry cell weight of 8.1 g/L, PHA content of 50%, and 3HV fraction of 20 mol%.

Consolidated bioprocessing for production of polyhydroxyalkanotes from red algae Gelidium amansii

Noncompetitive carbon sources such as algae are unconventional and promising raw material for sustainable biofuel production. The capability of one marine bacterium, Saccharophagus degradans 2-40 to degrade red seaweed Gelidium amansii for production of polyhydroxyalkanoates (PHA) was evaluated in this study. S. degradans can readily attach to algae, degrade algal carbohydrates, and utilize that material as main carbon source. Minimal media containing 8g/L G. amansii were used for the growth of S. degradans. The PHA content obtained was 17-27% of dry cell weight by pure culture of S. degradans and co-culture of S. degradans and Bacillus cereus, a contaminant found with S. degradans cultures. The PHA type was found to be poly(3-hydroxybutyrate) by gas chromatography and Fourier transform-infrared spectroscopy. This work demonstrates PHA production through consolidated bioprocessing of insoluble, untreated red algae by bacterial pure culture and co-culture.