Nanda Gopal Sahoo

Recent Trends of Polymer-Protein Conjugate Application in Biocatalysis: A Review

Proteins, particularly enzymes, are often used in bioreactors to catalyze biosynthetic reactions. However, one of the major challenges in applying proteins in bioreactors is enzyme stability and recovery. This review is an enlightening discussion on recent trends and advancements used to alleviate these limitations completely or to minimize them. Two major strategies, polymer-based enzyme immobilization and polymeric enzyme nanoreactors have been discussed systematically. Further, this review puts light on various methods such as smart polymer-based supports which have been exploited for their ability to regulate ligand-protein interactions. In addition, chemical moieties like protein-based microcrystals, cross-linked enzyme aggregates, and polymer-based nanoreactors have also been discussed in a comprehensive way focusing more on their unique applicability and target-specific actions. Polymer-based nanoreactors are described in detail in vivo together with enhancement of the enzyme stability and controlled function based on the compartmentalization of enzymes. Among the two novel nanoreactor approaches, dendrimers have been exploited as multifunctional enzymatic carriers, while capsomes are designed to regulate poly-enzymatic reactions through intravesicular compartmentalization of a multitude of enzymatic reactions.

An experimental modeling of trinomial bioengineering- crp, rDNA, and transporter engineering within single cell factory for maximizing two-phase bioreduction

A carbonyl reductase (cr) gene from Candida glabrata CBS138 has been heterologously expressed in cofactor regenerating E. coli host to convert Ethyl-4-chloro-3-oxobutanoate (COBE) into Ethyl-4-chloro-3-hydroxybutanoate (CHBE). The CR enzyme exhibited marked velocity at substrate concentration as high as 363mM with highest turnover number (112.77±3.95s-1). Solitary recombineering of such catalytic cell reproduced CHBE 161.04g/L per g of dry cell weight (DCW). Introduction of combinatorially engineered crp (crp*, F136I) into this heterologous E. coli host yielded CHBE 477.54g/L/gDCW. Furthermore, using nerolidol as exogenous cell transporter, the CHBE productivity has been towered to 710.88g/L/gDCW. The CHBE production has thus been upscaled to 8-12 times than those reported so far. qRT-PCR studies revealed that both membrane efflux channels such as acrAB as well as ROS scavenger genes such as ahpCF have been activated by engineering crp. Moreover, membrane protecting genes such as manXYZ together with solvent extrusion associated genes such as glpC have been upregulated inside mutant host. Although numerous proteins have been investigated to convert COBE to CHBE; this is the first approach to use engineering triad involving crp engineering, recombinant DNA engineering and transporter engineering together for improving cell performance during two-phase biocatalysis.

Genome mining, in silico validation and phase selection of a novel aldo-keto reductase from Candida glabrata for biotransformation

ABSTRACT Previously, we published cloning, overexpression, characterization and subsequent exploitation of a carbonyl reductase (cr) gene, belonging to general family aldo-keto reductase from Candida glabrata CBS138 to convert keto ester (COBE) to a chiral alcohol (ethyl-4-chloro-3-hydroxybutanoate or CHBE). Exploiting global transcription factor CRP, rDNA and transporter engineering, we have improved batch production of CHBE by trinomial bioengineering. Herein, we present the exploration of cr gene in Candida glabrata CBS138 through genome mining approach, in silico validation of its activity and selection of its biocatalytic phase. For exploration of the gene under investigation, 3 template genes were chosen namely Saccharomyces cerevisae YDR541c, YGL157w and YOL151w. The CR showed significant homology match, overlapping of substrate binding site and NADPH binding site with the template proteins. The binding affinity of COBE toward CR (−4.6 Kcal/ mol) was found higher than that of the template proteins (−3.5 to −4.5 Kcal/ mol). Biphasic biocatalysis with cofactor regeneration improved product titer 4∼5 times better than monophasic biotransformation. Currently we are working on DNA Shuffling as a next level of strain engineering and we demonstrate this approach herein as a future strategy of biochemical engineering.

Surface Modification of Carbon-Based Nanomaterials for Polymer Nanocomposites

Abstract Carbon nanomaterials such as graphene, carbon nanotubes, and fullerenes are superior materials over the other materials in the field of nanoscience and material science, due to their potential applications in various fields of science and technology. These materials are used as nanofiller for the preparation of polymer nanocomposites and are useful to enhance their properties exceptionally. However, their problem is associated with their poor solubility in aqueous and organic solvents which restricts their compatibility with polymer matrices, stopping these materials rfom achieving their full potential. To overcome this issue, surface modification of carbon nanomaterials via covalent and noncovalent functionalization with different polymers has previously been studied. This chapter highlights the noncovalent and covalent functionalization of carbon nanomaterials (graphene and CNT) through various types of interactions between polymers and carbon nanomaterials.

Electrical, thermal, and dielectric studies of ionic liquid-based polymer electrolyte for photoelectrochemical device

In this work, solution cast method was adapted for the preparation of 1-ethyl-3-methylimidazolium dicyanamide (EMImDCN)-doped solid polymer electrolyte. Optimum composition of polymer electrolyte (polyethylene oxide + sodium iodide) was treated as the host polymer. The ionic conductivity was further enhanced by adding low-viscosity ionic liquid (IL) EMImDCN. Electrical, thermal, dielectric, and photoelectrochemical properties of polymer host and IL-doped solid polymer electrolyte (ILDPE) are presented in detail. An electrochemical device, that is, dye-sensitized solar cell was fabricated using maximum conducting ILDPE film, which shows short-circuit current density of 0.118 mA/cm2, open-circuit voltage of 0.71 V, and overall efficiency of 0.061% at 1 sun condition.

Functionalized graphene oxides for drug loading, release and delivery of poorly water soluble anticancer drug: A comparative study

In this work, the modification of graphene oxides (GOs) have been done with hydrophilic and biodegradable polymer, polyvinylpyrrolidone (PVP) and other excipient β -cyclodextrin (β-CD) through covalent functionalization for efficient loading and compatible release of sparingly water soluble aromatic anticancer drug SN-38 (7-ethyl-10-hydroxy camptothecin). The drug was loaded onto both GO-PVP and GO-β-CD through the π-π interactions.The release of drug from both the nanocarriers were analyzed in different pH medium of pH 7 (water, neutral medium), pH 5 (acidic buffer) and pH 12 (basic buffer). The loading capacity and the cell killing activity of SN-38 loaded on functionalized GO were investigated comprehensively in human breast cancer cells MCF-7.Our findings shown that the cytotoxicity of SN-38 loaded to the polymer modified GO was comparatively higher than free SN-38. In particular, SN-38 loaded GO-PVP nanocarrier has more cytotoxic effect than GO-β-CD nanocarrier against MCF-7 cells, indicating that SN-38 loaded GO-PVP nanocarrier can be used as promising material for drug delivery and biological applications.

Schottky diodes from 2D germanane

We report on the fabrication and characterization of a Schottky diode made using 2D germanane (hydrogenated germanene). When compared to germanium, the 2D structure has higher electron mobility, an optimal band-gap, and exceptional stability making germanane an outstanding candidate for a variety of opto-electronic devices. One-atom-thick sheets of hydrogenated puckered germanium atoms have been synthesized from a CaGe2 framework via intercalation and characterized by XRD, Raman, and FTIR techniques. The material was then used to fabricate Schottky diodes by suspending the germanane in benzonitrile and drop-casting it onto interdigitated metal electrodes. The devices demonstrate significant rectifying behavior and the outstanding potential of this material.