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Immobilization of Xylanase Using a Protein-Inorganic Hybrid System
( Ashok Kumar ),( Sanjay K. S. Patel ),( Bharat Mardan ),( Raviteja Pagolu ),( Rowina Lestari ),( Seong-hoon Jeong ),( Taedoo Kim ),( Jung Rim Haw ),( Sang-yong Kim ),( In-won Kim ),( Jung-kul Lee ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.4
In this study, the immobilization of xylanase using a protein-inorganic hybrid nanoflower system was assessed to improve the enzyme properties. The synthesis of hybrid xylanase nanoflowers was very effective at 4°C for 72 h, using 0.25 mg/ml protein, and efficient immobilization of xylanase was observed, with a maximum encapsulation yield and relative activity of 78.5% and 148%, respectively. Immobilized xylanase showed high residual activity at broad pH and temperature ranges. Using birchwood xylan as a substrate, the V<sub>max</sub> and K<sub>m</sub> values of xylanase nanoflowers were 1.60 mg/ml and 455 μmol/min/mg protein, compared with 1.42 mg/ml and 300 μmol/min/mg protein, respectively, for the free enzyme. After 5 and 10 cycles of reuse, the xylanase nanoflowers retained 87.5% and 75.8% residual activity, respectively. These results demonstrate that xylanase immobilization using a proteininorganic hybrid nanoflower system is an effective approach for its potential biotechnological applications.
Otari, Sachin V.,Shinde, Vijay V.,Hui, Gao,Patel, Sanjay K.S.,Kalia, Vipin C.,Kim, In-Won,Lee, Jung-Kul Elsevier 2019 CERAMICS INTERNATIONAL Vol.45 No.5
<P><B>Abstract</B></P> <P>Noble-metal hybrid nanostructures have gained tremendous attention due to their potential roles in biomedical and catalytic applications. In this study, for the synthesis of silver nanoparticles (Ag NPs)–silica (SiO<SUB>2</SUB>) NPs, a novel green chemistry approach was employed, in which green tea biomolecule–encapsulated SiO<SUB>2</SUB> nanostructures were used for the reduction of silver ions to produce hybrid nanostructures within 300 s. The high-resolution transmission electron microscopy (HrTEM) revealed the formation of uniform ultrafine spherical Ag NPs that were evenly distributed in the nanostructures. The formed nanohybrid structures showed efficient catalytic activity for the formation of derivatives of dihydroquinoline, and retained 91% of their reusability capacity, even after 5 repeated cycles. Hence, this work provides a novel synthesis method not only for the synthesis of biomolecule-entrapped SiO<SUB>2</SUB> nanostructures, but also for the rapid formation of catalytically active hybrid nanostructures.</P>
Patel, Sanjay K.S.,Kumar, Virendra,Mardina, Primata,Li, Jinglin,Lestari, Rowina,Kalia, Vipin C.,Lee, Jung-Kul Elsevier 2018 Bioresource technology Vol.263 No.-
<P><B>Abstract</B></P> <P>In the present study, co-cultures of the methanotrophs <I>Methylocella tundrae</I>, <I>Methyloferula stellata</I>, and <I>Methylomonas methanica</I> were evaluated for improving methanol production with their application. Among the different combinations, the co-culture of <I>M. tundrae</I> and <I>M. methanica</I> increased methanol production to 4.87 mM using methane (CH<SUB>4</SUB>) as feed. When simulated biogas mixtures were used as feed, the maximum methanol production was improved to 8.66, 8.45, and 9.65 mM by free and encapsulated co-cultures in 2% alginate and silica-gel, respectively. Under repeated batch conditions, free and immobilized co-cultures using alginate and silica-gel resulted in high cumulative production, up to 24.43, 35.95, and 47.35 mM, using simulated biohythane (CH<SUB>4</SUB> and hydrogen), respectively. This is the first report of methanol production from defined free and immobilized co-cultures using simulated biogas mixtures as feed.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The co-culture of <I>Methanotrophs</I> is effective to produce methanol using simulated biogases. </LI> <LI> The silica-gel based encapsulated co-culture is an effective approach for methanol production. </LI> <LI> This is the first report of methanol production from immobilized co-cultures using biogas. </LI> </UL> </P>
Patel, Sanjay K.S.,Singh, Raushan K,Kumar, Ashok,Jeong, Jae-Hoon,Jeong, Seong Hun,Kalia, Vipin C.,Kim, In-Won,Lee, Jung-Kul Elsevier 2017 Bioresource technology Vol.241 No.-
<P><B>Abstract</B></P> <P>Biohythane may be used as an alternative feed for methanol production instead of costly pure methane. In this study, methanol production potential of <I>Methylocella tundrae</I> immobilized through covalent immobilization, adsorption, and encapsulation was evaluated. Cells covalently immobilized on groundnut shells and chitosan showed a relative methanol production potential of 83.9 and 91.6%, respectively, compared to that of free cells. The maximum methanol production by free cells and cells covalently immobilized on groundnut shells and chitosan was 6.73, 6.20, and 7.23mM, respectively, using simulated biohythane as a feed. Under repeated batch conditions of eight cycles, cells covalently immobilized on chitosan and groundnut shells, and cells encapsulated in sodium-alginate resulted in significantly higher cumulative methanol production of 37.76, 31.80, and 25.58mM, respectively, than free cells (18.57mM). This is the first report on immobilization of methanotrophs on groundnut shells and its application in methanol production using biohythane as a feed.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Biohythane is used as a feed to produce methanol by <I>Methylocella tundrae</I>. </LI> <LI> Compared to pure CH<SUB>4</SUB> as a feed, biohythane results in 1.9-fold higher methanol production. </LI> <LI> Covalently immobilized cells result in higher methanol production than free cells. </LI> <LI> Repeated use of the immobilized cells founds effective to improve methanol production. </LI> </UL> </P>
Production of Methanol from Methane by Encapsulated Methylosinus sporium
( Sanjay K. S. Patel ),( Jae-hoon Jeong ),( Sanjeet Mehariya ),( Sachin V. Otari ),( Bharat Madan ),( Jung Rim Haw ),( Jung-kul Lee ),( Liaoyuan Zhang ),( In-won Kim ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.12
Massive reserves of methane (CH<sub>4</sub>) remain unexplored as a feedstock for the production of liquid fuels and chemicals, mainly because of the lack of economically suitable and sustainable strategies for selective oxidation of CH4 to methanol. The present study demonstrates the bioconversion of CH<sub>4</sub> to methanol mediated by Type I methanotrophs, such as Methylomicrobium album and Methylomicrobium alcaliphilum. Furthermore, immobilization of a Type II methanotroph, Methylosinus sporium, was carried out using different encapsulation methods, employing sodium-alginate (Na-alginate) and silica gel. The encapsulated cells demonstrated higher stability for methanol production. The optimal pH, temperature, and agitation rate were determined to be pH 7.0, 30oC, and 175 rpm, respectively, using inoculum (1.5 mg of dry cell mass/ml) and 20% of CH<sub>4</sub> as a feed. Under these conditions, maximum methanol production (3.43 and 3.73 mM) by the encapsulated cells was recorded. Even after six cycles of reuse, the Na-alginate and silica gel encapsulated cells retained 61.8% and 51.6% of their initial efficiency for methanol production, respectively, in comparison with the efficiency of 11.5% observed in the case of free cells. These results suggest that encapsulation of methanotrophs is a promising approach to improve the stability of methanol production.
Patel, Sanjay K.S.,Anwar, Muhammad Z.,Kumar, Ashok,Otari, Sachin V.,Pagolu, Ravi T.,Kim, Sang-Yong,Kim, In-Won,Lee, Jung-Kul Elsevier 2018 Biochemical engineering journal Vol.132 No.-
<P><B>Abstract</B></P> <P>The structural morphology and composition of a support play a key role in the performance of nanoparticle-based enzymatic biosensors. In the present study, the influence of different functional groups, including glutaraldehyde, 3-aminopropyltriethoxysilane, carbodiimide, cyano, and polyethyleneimine for the immobilization of laccase on synthesized Fe<SUB>2</SUB>O<SUB>3</SUB> yolk-shell and commercially available Fe<SUB>2</SUB>O<SUB>3</SUB>, SrFe<SUB>12</SUB>O<SUB>19</SUB>, and Y<SUB>3</SUB>Fe<SUB>5</SUB>O<SUB>12</SUB> particles was analyzed. Glutaraldehyde-activated particles showed higher laccase activity after immobilization and higher relative detection currents for 2,6-dimethoxyphenol (2,6-DMP). The multi-shelled structural morphology of Fe<SUB>2</SUB>O<SUB>3</SUB> yolk-shell particles significantly improved the biosensing properties of immobilized laccase compared to that of spherical pure Fe<SUB>2</SUB>O<SUB>3</SUB> and composite SrFe<SUB>12</SUB>O<SUB>19</SUB> and Y<SUB>3</SUB>Fe<SUB>5</SUB>O<SUB>12</SUB> particles. The prepared biosensors showed high selectivity towards 2,6-DMP, with a sensitivity of 452 μA/mM/cm<SUP>2</SUP>. Under optimum conditions, the linear ranges of detection were as follows: 2,6-DMP (0.025–750 μM), guaiacol (0.10–250 μM), pyrogallol (0.25–250 μM), and 3,4-dihydroxy-<SMALL>L</SMALL>-phenylalanine (1.0–125 μM), with limit of detection values of 0.010, 0.052, 0.093, and 0.273 μM, respectively. Laccase immobilized on bio-friendly multi-shelled Fe<SUB>2</SUB>O<SUB>3</SUB> yolk-shell particles showed a broad linear range of detection, the lowest limit of detection, high sensitivity and stability, good reproducibility, anti-interference and recovery, and insignificant inhibition by laccase inhibitors.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fe<SUB>2</SUB>O<SUB>3</SUB> yolk-shell particles were used to prepare laccase biosensors. </LI> <LI> Particle composition and morphology exhibited significant variation in biosensing. </LI> <LI> Fe<SUB>2</SUB>O<SUB>3</SUB> yolk-shell biosensor showed a high selectivity towards 2,6-dimethoxyphenol. </LI> <LI> A broad linear range of detection with the lowest limit of detection (0.01 μM) was observed. </LI> </UL> </P>
( S. V. Otari ),( S. H. Pawar ),( Sanjay K. S. Patel ),( Raushan K. Singh ),( Sang-yong Kim ),( Jai Hyo Lee ),( Liaoyuan Zhang ),( Jung-kul Lee ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.4
A novel approach to synthesize silver nanoparticles (AgNPs) using leaf extract of Canna edulis Ker-Gawl. (CELE) under ambient conditions is reported here. The as-prepared AgNPs were analyzed by UV-visible spectroscopy, transmission emission microscopy, X-ray diffraction, Fourier transform-infrared spectroscopy, energy-dispersive analysis of X-ray spectroscopy, zeta potential, and dynamic light scattering. The AgNPs showed excellent antimicrobial activity against various pathogens, including bacteria and various fungi. The biocompatibility of the AgNPs was analyzed in the L929 cell line using NRU and MTT assays. Acridine orange/ ethidium bromide staining was used to determine whether the AgNPs had necrotic or apoptotic effects on L929 cells. The concentration of AgNPs required for 50% inhibition of growth of mammalian cells is far more than that required for inhibition of pathogenic microorganisms. Thus, CELE is a candidate for the eco-friendly, clean, cost-effective, and nontoxic synthesis of AgNPs.