Publications by authors named "Chinmay Hazra"

8 Publications

  • Page 1 of 1

Production, partial purification and characterization of a proteoglycan bioemulsifier from an oleaginous yeast.

Bioprocess Biosyst Eng 2020 Oct 14;43(10):1747-1759. Epub 2020 May 14.

Department of Biotechnology, Indian Institute of Technology Kharagpur, West Bengal, Kharagpur, 721302, India.

In this study, Meyerozyma caribbica, an indigenously isolated oleaginous yeast, produced in media containing glucose a bioemulsifier that was partially characterized as a proteoglycan based on preliminary analysis. Optimization of carbon:nitrogen (C:N) ratio revealed 30:1 as the suitable ratio for enhanced production. Apart from higher emulsification activity (E: 70-80%), this molecule showed strong emulsion stability over a wide range of pH (2.0-9.0), salinity (0.05%-10%, w/v) and temperature (- 80 °C to + 50 °C). The current study emphasizes on the determination of critical media parameters for improved and stable bioemulsifier production coupled with partial characterization and identification of the molecule. Thus, a proteoglycan-based bioemulsifier with such a stable emulsifying property can serve as a versatile and potential component in food, cosmetics and pharmaceutical formulations.
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http://dx.doi.org/10.1007/s00449-020-02361-1DOI Listing
October 2020

Biofunctionalized nanomaterials for in situ clean-up of hydrocarbon contamination: A quantum jump in global bioremediation research.

J Environ Manage 2020 Feb 10;256:109913. Epub 2019 Dec 10.

Department of Biotechnology, Indian Institute of Technology Kharagpur, West Bengal, 721302, India. Electronic address:

Interfacing organic or inorganic nanoparticles with biological entities or molecules or systems with the aim of developing functionalized nano-scale materials or composites for remediation of persistent organic hydrocarbon pollutants (such as monocyclic and polycyclic aromatic hydrocarbons, MAH/PAH) has generated great interest and continues to grow almost unabated. However, the usefulness and potency of these materials or conjugates hinges over several key barriers, including structural assembly with fine-tuned control over nanoparticle/biomolecule ratio, spatial orientation and activity of biomolecules, the nano/bio-interface strategy and hierarchical architecture, water-dispersibility and long term colloidal stability in environmental media, and non-specific toxicity. The present review thus critically analyses, discusses and interprets recently reported attempts and approaches to functionalize nanoparticles with biomolecules. Since there is no comprehensive and critical reviews on the applications of nanotechnology in bioremediation of MAHs/PAHs, this overview essentially captures the current global scenario and vision on the use and future prospects of biofunctionalized nanomaterials with respect to their strategic interactions involved at the nano/bio-interface essential to understand and decipher the structural and functional relationships and their impact on persistent hydrocarbon remediation.
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http://dx.doi.org/10.1016/j.jenvman.2019.109913DOI Listing
February 2020

A strategic approach of enzyme engineering by attribute ranking and enzyme immobilization on zinc oxide nanoparticles to attain thermostability in mesophilic Bacillus subtilis lipase for detergent formulation.

Int J Biol Macromol 2019 Sep 8;136:66-82. Epub 2019 Jun 8.

Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India. Electronic address:

The present study envisaged rationalized protein engineering approach to attain thermostability in a mesophilic Bacillus subtilis lipase. Contributing amino acids for thermostability were analyzed from homologous thermophilic-mesophilic protein dataset through relative abundance and generated ranking model. Analyses divulged priority of charged amino acids for thermostability. Ranking model was used to predict thermostabilizing mutations. Three lipase mutants, bsl_the1 (V149K, Q150E), bsl_the2 (F41K, W42E, V149K, Q150E) and bsl_the3 (F41K, W42E, P119E, Q121K, V149K, Q150E) were generated and validated through in silico and in vitro approaches for improved activity and thermostability. ZnO nanoparticles were synthesized by precipitation method and functionalized using polyethylenimine, APTES and glutaraldehyde for lipase immobilization. The immobilization was confirmed through various analytical techniques. Analysis revealed bsl_wt showed optimum activity at 35 °C and pH 8 which was increased to 60 °C and pH 10 in case of ZnO-bsl_the3. The ZnO-bsl_the3 showed 80% of their initial activity after 60 days of storage stability and retained 78% of activity after 20 cycles of reuse. Lipases were applied for oil and grease stain removal from fabric. ZnO-bsl_the3 removed 90% and 82% of oil and grease stains, respectively. Conclusively, it revealed a promising perspective of low-cost nanobiocatalysts in detergent formulation.
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http://dx.doi.org/10.1016/j.ijbiomac.2019.06.042DOI Listing
September 2019

Electrodeposited functionally graded coating inhibits Gram-positive and Gram-negative bacteria by a lipid peroxidation mediated membrane damage mechanism.

Mater Sci Eng C Mater Biol Appl 2019 Sep 29;102:623-633. Epub 2019 Apr 29.

School of Nanoscience and Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India; Department of Metallurgical and Materials Engineering, Indian Institute of Technology Kharagpur, Kharagpur 721302, India. Electronic address:

The current work deals with a time-dependent study to track the antibacterial action of electrodeposited Cu, Cu-SiC functionally graded coating (FGC) against Escherichia coli NCIM 2931 (Gram-negative) and Bacillus subtilis NCIM 2063 (Gram-positive). After 24 h of incubation, the Cu, Cu-SiC FGC causes 7 Escherichia coli NCIM 2931 and 10 Bacillus subtilis NCIM 2063 log reduction of planktonic cells. The outer membrane permeabilization experiment proves that the intake of excessive Cu ions leads to the damage of bacterial cell membrane followed by lipid degradation. The thiobarbituric acid reactive substances assay reveals that Cu ions released from the surface of Cu, Cu-SiC FGC triggers the oxidative degeneration of phospholipids (most abundant constituent of bacterial cell membrane). This was further cross-verified using atomic absorption spectroscopy. From 0 to 24 h, the bacterial morphology is characterized using transmission electron microscope and scanning electron microscope which shows the cytoplasmic leakage and cell death. The Cu, Cu-SiC FGC also exhibits hydrophobic surface (contact angle of 144°) which prevents the bacterial adherence to the surface and thus, inhibits them to penetrate into its bulk. The observed results of antibacterial and anti-adhesion properties of Cu, Cu-SiC FGC are compared with single-layered metallic Cu and Cu-SiC nanocomposite coatings. Hence, the electrodeposited Cu, Cu-SiC FGC has the potential to serve as an inexpensive touch surface alternative for the healthcare industries.
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http://dx.doi.org/10.1016/j.msec.2019.04.087DOI Listing
September 2019

Statistical modeling and optimization of culture conditions by response surface methodology for 2,4- and 2,6-dinitrotoluene biodegradation using Rhodococcus pyridinivorans NT2.

3 Biotech 2016 Dec 19;6(2):155. Epub 2016 Jul 19.

School of Life Sciences, North Maharashtra University, Jalgaon, 425 001, Maharashtra, India.

To improve biodegradability (% biodegradation) and specific growth rate of Rhodococcus pyridinivorans NT2, culture medium and environmental parameters were screened and optimized using the statistical design techniques of Plackett-Burman and response surface methodology. Of the process variables screened, DNTs (2,4-DNT and 2,6-DNT), MgSO·7HO, temperature and inoculum size (O.D.) were selected as the most important (P value <0.05) factors. In multiresponse analysis of central composite design, medium formulation consisting of 474/470 mg l 2,4-DNT/2,6-DNT, 0.11 g l MgSO·7HO, 37.5 °C temperature and 1.05 OD inoculum size were found to predict maximum % degradation and specific growth rate of 97.55 % and 0.19 h, respectively. The validity of the optimized variables was verified in shake flasks. The optimized media significantly shortened the time required for biodegradation of DNTs while providing a nearly 30 % (for 2,4-DNT) and 70 % (for 2,6-DNT) increased biodegradation along with 5.64-fold increase in specific growth rate for both DNTs.
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http://dx.doi.org/10.1007/s13205-016-0468-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4951380PMC
December 2016

Extracellular biosynthesis of zinc oxide nanoparticles using Rhodococcus pyridinivorans NT2: multifunctional textile finishing, biosafety evaluation and in vitro drug delivery in colon carcinoma.

J Photochem Photobiol B 2014 Nov 12;140:194-204. Epub 2014 Aug 12.

University Institute of Chemical Technology, North Maharashtra University, Jalgaon, Maharashtra, India.

In this study, zinc oxide (ZnO) nanoparticles (NPs) were rapidly synthesized from zinc sulfate solution at room temperature using a metabolically versatile actinobacteria Rhodococcus pyridinivorans NT2. The morphology, structure and stability of the synthesized ZnO NPs were studied using UV-visible absorption spectroscopy, X-ray Diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, field emission scanning electron microscopy (FESEM) with energy dispersive spectroscopy (EDS), transmission electron microscopy (TEM), Zeta potential, and thermogravimetry. The data indicated that the synthesized nanoparticles were moderately stable, hexagonal phase, roughly spherical with average particle diameter in the range of 100-120 nm. Results obtained on examination of protein expression revealed that cell enzymes and extracellular protein systems of Rhodococcus sp. may take part in synthesis process. Furthermore, the ZnO NPs were coated onto textile fabrics to enhance UV-blocking, self-cleaning and antibacterial properties. Ultraviolet protecting factor (UPF) indicating UV-blocking properties of ZnO NPs coated textile fabrics were determined as 65, 88, 121, 172 and 241 for 1, 2, 3, 4 and 5 gm(-2) of ZnO NPs, respectively. Besides, self-cleaning activity was assessed by investigating photocatalytic activity on malachite green as well as antibacterial activity against aerobic Gram-positive Staphylococcus epidermidis NCIM 2493 (ATCC 12228). The antibacterial effects of these textiles were evaluated using ISO 20743 standard. In addition, ZnO NPs exhibited a preferential ability to kill HT-29 cancerous cells as compared with normal peripheral blood mononuclear cells (PBMCs).
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http://dx.doi.org/10.1016/j.jphotobiol.2014.08.001DOI Listing
November 2014

Ultrasound-assisted/biosurfactant-templated size-tunable synthesis of nano-calcium sulfate with controllable crystal morphology.

Ultrason Sonochem 2014 May 30;21(3):1117-31. Epub 2013 Dec 30.

University Institute of Chemical Technology, North Maharashtra University, Jalgaon 425 001, India. Electronic address:

Nano-sized crystals of alpha calcium sulfate hemihydrate (α-HH) with considerable morphology-dependent properties find promising applications in the clinical fields as a cementitious material. Towards this end, ultrasound-assisted rhamnolipid and surfactin biosurfactant-template route is explored to control the morphology and aspect ratio of nano-CaSO4 by adjusting the mass ratio of rhamnolipid/H2O, surfactin/H2O and rhamnolipid/surfactin. The change in the molar ratio of [SO4(2-)]:[Ca(2+)] results in modification in variable morphology and size of nano-CaSO4 including long, short rods and nanoplates. With increase in the rhamnolipid/H2O ratio from 1.3 to 4.5, the crystal length decreases from 3 μm to 600 nm with the corresponding aspect ratio reduced sharply from 10 to 3. Similarly, the crystal morphology gradually changes from submicrometer-sized long rod to hexagonal plate, and then plate-like appearance with increase in surfactin concentration. The preferential adsorption of rhamnolipid on the side facets and surfactin on the top facets contributes to the morphology control. The process using 50% amplitude with a power input of 45.5 W was found to be the most ideal as observed from the high yields and lower average l/w aspect ratio, leading to more than 94% energy savings as compared to that utilized by the conventional process. As a morphology and crystal habit modifier, effects of Mg(2+) and K(+) ions on α-HH growth were investigated to find an optimal composition of solution for α-HH preparation. Mg(2+) ions apparently show an accelerating effect on the α-HH growth; however, the nucleation of α-HH is probably retarded by K(+) ions. Thus, the present work is a simple, versatile, highly efficient approach to controlling the morphology of α-HH and thereby, offers more opportunities for α-HH multiple applications.
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http://dx.doi.org/10.1016/j.ultsonch.2013.12.020DOI Listing
May 2014

Biodegradation of 4-nitrotoluene with biosurfactant production by Rhodococcus pyridinivorans NT2: metabolic pathway, cell surface properties and toxicological characterization.

Biodegradation 2013 Nov 7;24(6):775-93. Epub 2013 Feb 7.

School of Life Sciences, North Maharashtra University, Jalgaon, 425 001, Maharashtra, India.

A novel 4-nitrotoluene-degrading bacterial strain was isolated from pesticides contaminated effluent-sediment and identified as Rhodococcus pyridinivorans NT2 based on morphological and biochemical properties and 16S rDNA sequencing. The strain NT2 degraded 4-NT (400 mg l(-1)) with rapid growth at the end of 120 h, reduced surface tension of the media from 71 to 29 mN m(-1) and produced glycolipidic biosurfactants (45 mg l(-1)). The biosurfactant was purified and characterized as trehalose lipids. The biosurfactant was stable in high salinity (10 % w/v NaCl), elevated temperatures (120 °C for 15 min) and a wide pH range (2.0-10.0). The noticeable changes during biodegradation were decreased hydrophobicity; an increase in degree of fatty acid saturation, saturated/unsaturated ratio and cyclopropane fatty acid. Biodegradation of 4-NT was accompanied by the accumulation of ammonium (NH4 (+)) and negligible amount of nitrite ion (NO2 (-)). Product stoichiometry showed a carbon (C) and nitrogen (N) mass balance of 37 and 35 %, respectively. Biodegradation of 4-NT proceeded by oxidation at the methyl group to form 4-nitrobenzoate, followed by reduction and hydrolytic deamination yielding protocatechuate, which was metabolized through β-ketoadipate pathway. In vitro and in vivo acute toxicity assays in adult rat (Rattus norvegicus) showed sequential detoxification and the order of toxicity was 4-NT >4-nitrobenzyl alcohol >4-nitrobenzaldehyde >4-nitrobenzoate >> protocatechuate. Taken together, the strain NT2 could be used as a potential bioaugmentation candidate for the bioremediation of contaminated sites.
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http://dx.doi.org/10.1007/s10532-013-9627-4DOI Listing
November 2013
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