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Enzyme and Microbial Technology

Paula Nicolás, Verónica L Lassalle, María L Ferreira
The aim of this manuscript was to study the application of a new method of protein quantification in Candida antarctica lipase B commercial solutions. Error sources associated to the traditional Bradford technique were demonstrated. Eight biocatalysts based on C. antarctica lipase B (CALB) immobilized onto magnetite nanoparticles were used. Magnetite nanoparticles were coated with chitosan (CHIT) and modified with glutaraldehyde (GLUT) and aminopropyltriethoxysilane (APTS). Later, CALB was adsorbed on the modified support...
February 2017: Enzyme and Microbial Technology
Jong-Min Park, Jo-Il Kim, Joo-Yoon Noh, Mira Kim, Min-Jung Kang, Jae-Chul Pyun
Many kinds of susceptibility test for β-lactam antibiotics have been used to determine the antibiotic resistance of bacterial strains. Here, a sensitive antibiotic susceptibility test was presented by using a specialized reaction tool for laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS) based on parylene-matrix chip. The β-lactamase assay was carried out in a specialized reaction tool by (1) concentrating the bacterial strain and (2) incubating the bacteria with penicillin-G. The parylene-matrix chip was produced by deposition of a partially porous parylene-N thin film on a dried organic matrix array, and the products of β-lactamase reaction in the low range of mass-to-charge ratio (m/z<500) could be effectively analyzed by using a parylene-matrix chip...
February 2017: Enzyme and Microbial Technology
Weixiong Hu, Xiaoyun Liu, Yufeng Li, Daling Liu, Zhihe Kuang, Chuiwen Qian, Dongsheng Yao
β-Mannanase has been widely used in industries such as food and feed processing and thus has been a target enzyme for biotechnological development. In this study, we sought to improve the stability and protease resistance of a recombinant β-mannanase, MAN47 from Armillariella tabescens, through rationally designed N-glycosylation. Based on homology modeling, molecular docking, secondary structure analysis and glycosylation feasibility analysis, an enhanced aromatic sequon sequence was introduced into specific MAN47 loop regions to facilitate N-glycosylation...
February 2017: Enzyme and Microbial Technology
Wei Xiong, Jiang-Ke Yang, Fang-Yuan Chen, Zheng-Gang Han
The cellulase-mediated degradation of cellulosic materials, which is initiated by endoglucanases by the random cleavage of the glycosidic bonds between glucose units to break long cellulose molecules into shorter ones, represents a major carbon flow in the global carbon cycle. The structure of a typical endoglucanase contains a classical (α/β)8 barrel fold catalytic domain, a linker region and a cellulose-binding domain. In this study, we found that both the full-length enzyme and the catalytic domain of endoglucanase EGL1 cloned from Penicillium crustosum strain 601 have CMCase and FPase activity...
February 2017: Enzyme and Microbial Technology
Taku Orita, Makiko Sakka, Tetsuya Kimura, Kazuo Sakka
Three cellulosomal subunits of Ruminiclostridium josui, the full-length scaffolding protein CipA (RjCipA), a cellulase Cel5B (RjCel5B) and a xylanase Xyn10C (RjXyn10C), were successfully produced by Escherichia coli recombinant clones. RjCel5B and RjXyn10C were characterized as an endoglucanase and an endoxylanase, respectively. RjCipA, RjCel5B and Xyn10C adsorbed to microcrystalline cellulose (Funacel) and rice straw powder. Interaction between RjCel5B and RjCipA, and RjXyn10C and RjCipA were confirmed by qualitative assays...
February 2017: Enzyme and Microbial Technology
M Shivakumar, K L Nagashree, S Yallappa, S Manjappa, K S Manjunath, M S Dharmaprakash
Herein, we described for the first time, biosynthesis of silver nanoparticles (AgNPs) using pre-hydrolyzed liquor of Eucalyptus wood under ambient conditions. The pre-hydrolyzed liquor containing a high amount of metabolites such as polyphenols, hemicelluloses and its derivatives are mainly assisted for the reduction and stabilization process of Ag+ ions to AgNPs. The formation of AgNPs is monitored by recording the UV-vis spectrophotometer for surface plasmon resonance (SPR) peak observed at ∼415nm. The intensity of SPR increased linearly with increasing the reaction time at ambient condition...
February 2017: Enzyme and Microbial Technology
Karen O Osiro, Brenda R de Camargo, Rachel Satomi, Pedro Ricardo V Hamann, Jéssica Pinheiro Silva, Marcelo Valle de Sousa, Betania F Quirino, Elaine N Aquino, Carlos R Felix, André Melro Murad, Eliane F Noronha
The main goal of the present study was a complete proteomic characterization of total proteins eluted from residual substrate-bound proteins (RSBP), and cellulosomes secreted by Clostridium thermocellum B8 during growth in the presence of microcrystalline cellulose as a carbon source. The second goal was to evaluate their potential use as enzymatic blends for hydrolyzing agro-industrial residues to produce fermentable sugars. Protein identification through LC-MS/MS mass spectrometry showed that the RSBP sample, in addition to cellulosomal proteins, contains a wide variety of proteins, including those without a well-characterized role in plant cell wall degradation...
February 2017: Enzyme and Microbial Technology
Thien-Kim Le, Hyun-Hee Jang, Ha Thi Huong Nguyen, Tiep Thi My Doan, Ga-Young Lee, Ki Deok Park, Taeho Ahn, Young Hee Joung, Hyung-Sik Kang, Chul-Ho Yun
Enzymatic conversion of natural glycosides to their corresponding hydroxylated products using cytochromes P450 has significant advantages over synthetic chemistry and even enzyme-catalyzed glycosylation of chemicals. At present, the basic strategy for making glycosides of stilbenoid compounds is to use the glycosylation activity of enzymes, such as glycosyltransferases. Here, an efficient synthesis of a valuable (E)-astringin, a piceatannol glucoside, was developed using CYP102A1 via the highly regioselective C-3' hydroxylation of polydatin, a resveratrol glucoside...
February 2017: Enzyme and Microbial Technology
Manisha J Patel, Rekha C Akhani, Arti T Patel, Samir R Dedania, Darshan H Patel
l-ribose and d-tagatose are biochemically synthesized using sugar isomerases. The l-arabinose isomerase gene from Shigella flexneri (Sf-AI) was cloned and expressed in Escherichia coli BL-21. Sf-AI was applied for the bioproduction of d-tagatose from d-galactose. l-ribose synthesis was performed by two step isomerization using Sf-AI and d-lyxose/ribose isomerase from Cohnella laevoribosii. The overall 22.3% and 25% conversion rate were observed for d-tagatose and l-ribose production from d-galactose and l-arabinose respectively...
February 2017: Enzyme and Microbial Technology
Gökçe Şeker Karatoprak, Gamze Aydin, Berrak Altinsoy, Cevahir Altinkaynak, Müberra Koşar, Ismail Ocsoy
Herein, we report the biosynthesis of Ag NPs, for the first time, using identified antimicrobial molecules (gallic acid+apocynin) and (gallic acid+apocynin+quercetin) from the medicinal plant Pelargonium endlicherianum Fenzl. and dramatically enhanced antimicrobial activity. We also investigate the role of each molecule on formation Ag NPs and explain the increase in the antimicrobial activity of identified molecules mediated Ag NPs. The extraction protocols, 11% ethanol and 70% methanol, resulted in identification of different constituents of gallic acid+apocynin (M1) and gallic acid+apocynin+quercetin (M2) with respective concentrations...
February 2017: Enzyme and Microbial Technology
Veronica da Silva Ferreira, Mateus Eugenio ConzFerreira, Luís Maurício T R Lima, Susana Frasés, Wanderley de Souza, Celso Sant'Anna
Silver nanoparticles are powerful antimicrobial agents. Here, the synthesis of silver chloride nanoparticles (AgCl-NPs) was consistently evidenced from a commercially valuable microalgae species, Chlorella vulgaris. Incubation of C. vulgaris conditioned medium with AgNO3 resulted in a medium color change to yellow/brown (with UV-vis absorbance at 415nm), indicative of silver nanoparticle formation. Energy-dispersive X-ray spectroscopy (EDS) of purified nanoparticles confirmed the presence of both silver and chlorine atoms, and X-ray diffraction (XRD) showed the typical pattern of cubic crystalline AgCl-NPs...
February 2017: Enzyme and Microbial Technology
Rhudith B Cabulong, Kris Niño G Valdehuesa, Kristine Rose M Ramos, Grace M Nisola, Won-Keun Lee, Chang Ro Lee, Wook-Jin Chung
The microbial production of renewable ethylene glycol (EG) has been gaining attention recently due to its growing importance in chemical and polymer industries. EG has been successfully produced biosynthetically from d-xylose through several novel pathways. The first report on EG biosynthesis employed the Dahms pathway in Escherichia coli wherein 71% of the theoretical yield was achieved. This report further improved the EG yield by implementing metabolic engineering strategies. First, d-xylonic acid accumulation was reduced by employing a weak promoter which provided a tighter control over Xdh expression...
February 2017: Enzyme and Microbial Technology
Yihai Li, Gaku Arakawa, Gaku Tokuda, Hirofumi Watanabe, Manabu Arioka
A β-glucosidase (BG), PaBG1b, from the xylophagous cockroach Panesthia angustipennis spadica was heterologously expressed in the methylotrophic yeast Pichia pastoris, purified, and biochemically characterized. Post-translational modification and N-terminal sequencing analysis demonstrated that the expression product was comprised of two polypeptides with different N-terminal sequences, presumably due to the presence of lysine-arginine (KR) sequence in the putative mature region. Substrate specificity analysis showed that PaBG1b hydrolyzed a broad range of substrates including cellohexaose, with the preference for aryl β-d-fucosyl linkage and laminaribiose...
February 2017: Enzyme and Microbial Technology
Usman Liaqat, Hyuk Ko, Hwal Suh, Misu Lee, Jae-Chul Pyun
PC-12 cells originate from neuroblastic cells, which have an ability to differentiate into neuronlike cells. In this work, the purpose was to estimate the influence of microenvironments on cell attachment and neuritogenesis capacity of PC-12 cells on parylene-N and parylene-C films with and without ultraviolet (UV) light treatment. The estimate of total cell number after incubation for 72h, the ratio of adherent to suspended cells, counting of neurite outgrowths on parylene-N or parylene-C films after UV exposure suggested that these films were suitable for proliferation as well as differentiation of PC-12 cells...
February 2017: Enzyme and Microbial Technology
Zheng-Yu Yan, Qing-Qing Du, Jing Qian, Dong-Yu Wan, Sheng-Mei Wu
In the paper, a green and efficient biosynthetical technique was reported for preparing cadmium sulfide (CdS) quantum dots, in which Escherichia coli (E. coli) was chosen as a biomatrix. Fluorescence emission spectra and fluorescent microscopic photographs revealed that as-produced CdS quantum dots had an optimum fluorescence emission peak located at 470nm and emitted a blue-green fluorescence under ultraviolet excitation. After extracted from bacterial cells and located the nanocrystals' foci in vivo, the CdS quantum dots showed a uniform size distribution by transmission electron microscope...
January 2017: Enzyme and Microbial Technology
Seo-Yoon Chang, Ji-Hong Bong, Gu Yoo, Misu Lee, Min-Jung Kang, Joachim Jose, Jae-Chul Pyun
The autodisplay technology has been applied for expression of a desired protein on the outer membrane (OM) of Escherichia coli. In this work, the OM fractions of E. coli with two autodisplayed proteins were separately prepared and mixed to demonstrate the feasibility of control over the ratio of two autodisplayed proteins. As the first model, Z-domain and streptavidin were autodisplayed, and their activities were tested by means of the combined OM layer in a 96-well microplate and a surface plasmon resonance (SPR) biosensor...
January 2017: Enzyme and Microbial Technology
Junjarus Sermsathanaswadi, Sirilak Baramee, Chakrit Tachaapaikoon, Patthra Pason, Khanok Ratanakhanokchai, Akihiko Kosugi
A newly isolated endo-β-1,4-xylanase (Xyn10E) from Paenibacillus curdlanolyticus B-6 has a modular structure consisting of a family 22 carbohydrate-binding module (CBM), a glycoside hydrolase (GH) family 10 catalytic domain, two fibronectin type III (Fn3) domains, and a family 3 CBM at the C-terminus. Intact Xyn10E (rXyn10E), CBM22-deleted Xyn10E (X-CBM3), CBM3-deleted Xyn10E (X-CBM22), and GH10 catalytic domain only (X-GH10) were expressed in Escherichia coli. rXyn10E showed bifunctional degradation activity toward xylan and β-glucan and also degraded microcrystalline cellulose...
January 2017: Enzyme and Microbial Technology
Fidal V T K P, Saikumar Inguva, Satheesh Krishnamurthy, Enrico Marsili, Jean-Paul Mosnier, Chandra T S
Al doped and undoped ZnO thin films were deposited by pulsed-laser deposition on polycarbonate sheets. The films were characterized by optical transmission, Hall effect measurement, XRD and SEM. Optical transmission and surface reflectometry studies showed good transparency with thicknesses ∼100nm and surface roughness of 10nm. Hall effect measurements showed that the sheet carrier concentration was -1.44×10(15)cm(-2) for AZO and -6×10(14)cm(-2) for ZnO. The films were then modified by drop-casting glucose oxidase (GOx) without the use of any mediators...
January 2017: Enzyme and Microbial Technology
Somaieh Rahnama, Behnaz Saffar, Zahra Fanaei Kahrani, Mahboobeh Nazari, Rahman Emamzadeh
Renilla luciferase is a bioluminescent enzyme which is broadly used as a reporter protein in molecular biosensors. In this study, a novel luciferase with desired light emission wavelength and thermostability is reported. The results indicated that the new luciferase, namely super RLuc8, had a red-shifted spectrum and showed stable light emission. Super RLuc8 showed a 10-fold (p-value=0.0084) increase in the thermostability at 37°C after 20min incubation, in comparison to the native enzyme. The optimum temperature of the mutant increased from 30 to 37°C...
January 2017: Enzyme and Microbial Technology
Mojtaba Mortazavi, Saman Hosseinkhani
Protein engineering can provide useful approaches for loop anchoring and mutation of surface-exposed loop residues to Arg for the design of thermostable proteins. In this context and due to the high proportion of surface loops, some of the solvent-exposed residues in the Lampyris turkestanicus luciferase were mutated to Arg. Using the red-emitter mutant luciferase (E354R/Arg356), the single (-Q35R, -I182R, -I232R and -L300R), double (-Q35R/I232R) and triple (-Q35R/I232R/I182R) mutant luciferases were introduced...
January 2017: Enzyme and Microbial Technology
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