• Journal of Microbiology and Biotechnology
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• v.30 no.9
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• pp.1436-1442
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• 2020

Amylosucrase (ASase, E.C. 2.4.1.4) is capable of efficient glucose transfer from sucrose, acting as the sole donor molecule, to various functional acceptor compounds, such as polyphenols and flavonoids. An ASase variant from Deinococcus geothermalis, in which the 226^th alanine is replaced with asparagine (DgAS-A226N), shows increased polymerization activity due to changes in the flexibility of the loop near the active site. In this study, we further investigated how the mutation modulates the enzymatic activity of DgAS using molecular dynamics and docking simulations to evaluate interactions between the enzyme and phenolic compounds. The computational analysis revealed that the A226N mutation could induce and stabilize structural changes near the substrate-binding site to increase glucose transfer efficiency to phenolic compounds. Kinetic parameters of DgAS-A226N and WT DgAS were determined with sucrose and 4-methylumbelliferone (MU) as donor and acceptor molecules, respectively. The k_cat/K_m value of DgAS-A226N with MU (6.352 mM^-1min^-1) was significantly higher than that of DgAS (5.296 mM^-1min^-1). The enzymatic activity was tested with a small phenolic compound, hydroquinone, and there was a 1.4-fold increase in α-arbutin production. From the results of the study, it was concluded that DgAS-A226N has improved acceptor specificity toward small phenolic compounds by way of stabilizing the active conformation of these compounds.

• Journal of Ginseng Research
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• v.14 no.1
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• pp.57-62
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• 1990

In order to Investigate the mechanism of the leaf-burning disease of ginseng (Panax ginseng C.A. Meyer), studies on the generation of singlet oxygen (1O2) and the photooxidation of the pigments were carried out in comparison with the ones of soybean (G1ycine max L). The studies were mainly focalized on the effects of light intensity, light intensity, inhibitor and electron donor/acceptor of the Photosynthetic electron transport system. When we measured the amounts of 1O2 generated in the thylakoids of ginseng and soybean by the irradiation of light (300 w/m2) as a function its time. It was identified that a higher amount of 1O2 was formed in the ginseng thylakoid than the case of soybean. A generation ratio of lO2 between ginseng and soybean sltbstantially identical in the range of light intensities 50∼150w/m2 However much higher amount of 1O2 was generated in ginseng by irradiation of strong intensity of light (200 500w/m2). Wave length dependency on the generation of 1O2 and the pigment photooxidation was observed on ginseng thylakoids; red light (600-700 nm) gave a maximum effect in the contrast with blur green light (400-60 nm). When the ginseng thylalioid was treated with the electron donor (Mn2+) and acceptors (DCPIP, FeCy) of the photosynthetic electron transport system. a drastic inhibition of 1O2 generation was observed. However, treatment with its inhibitors (DCMU, KCW) activated 1O2 generation. An interesting fact that an electron donor or acceptor of the photosystem II(P680) Inhibited 1O2 generation, suggests an intimate relationship between 1O2 generation and photosystem II.

• Journal of Photoscience
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• v.9 no.2
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• pp.385-387
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• 2002

Direct EPR evidence of the photo-generation of superoxide radicals ( $O_2$$^{-.}$) was obtained by using spin trapping techniques in spinach photosystem II (PSII) membranes. $O_2$$^{-.}$ was detected by following the formation of 5-diethoxyphosphoryl-5-methyl-1 -pyrroline-N-oxide (DEPMPO) superoxide adducts, DEPMPO-OOH. The significant increase of the EPR signal amplitude of DEPMPO-OOH in N$H_2O$H-, CaC $l_2$- and NaCl-treated PSII membranes showed that the oxygen-evolving system has a close relation to the $O_2$$^{-.}$ production. PSII membranes with inactivated donor side could not prevent the $O_2$$^{-.}$ production efficiently. Treatments on PSII donor side also influence the maximum level and the kinetics of Chlorophyll (Chi) a fluorescence. Results suggested that manganese cluster and extrinsic proteins might affect Chi a fluorescence in ways different from that happens at the acceptor side of PSII.SII.SII.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1845-1854
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• 2016

The transglycosylation activity of amylosucrase (ASase) has received significant attention owing to its use of an inexpensive donor, sucrose, and broad acceptor specificity, including glycone and aglycone compounds. The transglycosylation reaction of recombinant ASase from Deinococcus radiopugnans (DRpAS) was investigated using various phenolic compounds, and quercetin-3-O-rutinoside (rutin) was found to be the most suitable acceptor molecule used by DRpAS. Two amino acid residues in DRpAS variants (DRpAS Q299K and DRpAS Q299R), assumed to be involved in acceptor binding, were constructed by site-directed mutagenesis. Intriguingly, DRpAS Q299K and DRpAS Q299R produced 10-fold and 4-fold higher levels of rutin transglycosylation product than did the wild-type (WT) DRpAS, respectively. According to in silico molecular docking analysis, the lysine residue at position 299 in the mutants enables rutin to more easily position inside the active pocket of the mutant enzyme than in that of the WT, due to conformational changes in loop 4.

• Macromolecular Research
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• v.16 no.5
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• pp.446-456
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• 2008

A recent theoretical approach based on the coupling of both the Flory-Huggins (FH) and the Association Equilibria thermodynamic (AET) theories was modified and adapted to study the miscibility properties of a multi-component system formed by two polymers (a proton-donor and a proton-acceptor) and a proton-acceptor solvent, named copolymer(A)/solvent(B)/polymer(C). Compatibility between polymers was mainly attained by hydrogen-bonding between the hydroxyl group on the phenol unit of the poly(styrene-co-vinyl phenol) (PSVPh) and the carbonyl group of the biodegradable and environmentally friendly poly(3-hydroxybutyrate) (PHB). However, the self-association of PSVPh and specific interactions between the PSVPh and the H-acceptor group (an ether oxygen atom) of the epichlorohydrin (ECH) solvent were also established in a lower extension, which competed with the polymer-polymer association. All the binary specific interactions and their dependence with the system composition as well as with the copolymer content were evaluated and quantified by means of two excess functions of the Gibbs tree energy, ${\Delta}g_{AB}$ and ${\Delta}g_{AC}$. Experimental results from fluorescence spectroscopy were consistent with the theoretical simulations derived with the model, which could also be applied and extended to predict the miscibility in solution of any polymer blend with specific interactions.

• Archives of Pharmacal Research
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• v.24 no.4
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• pp.316-322
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• 2001

Arylsulfate sulfotransferase (ASST) transfers a sulfate group from a phenolic sulfate ester to a phenolic acceptor substrate. In the present study, the gene encoding ASST was cloned from a genomic library copy of Citrobacter freundii, subcloned into the vector pGEM3Zf(-) and sequenced. Sequencing revealed two contiguous open reading frames (ORF1 and ORF2) on the same strand and based on amino acid sequence homologyl they were designated as astA and dsbA, respectively. The amino acid sequence of astA deduced from C. freundii was highly similar to that of the Salmonella typhimurium, Enterobacter amnigenus, Klebsiella, Pseudomonas putida, and Campylobacter jejuni, encoded by the astA genes. However, the ASST activity assay revealed different acceptor specificities. Using p-nitrophenyl sulfate (PNS) as a donor substrate, $\alpha$-naphthol was found to be the best acceptor substrate, followed by phenol, resorcinol, p-acetaminophen, tyramine and tyrosine.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.127-131
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• 1999

Anaerobically grown cells of an Fe(III)-reducing bacterium, Shewanella putrefaciens IR-l, were electrochemically active with an apparent reduction potential of about 0.15 V against a saturated calomel electrode in the cyclic voltammetry. The bacterium did not grow fermentatively on lactate, but grew in an anode compartment of a three-electrode electrochemical cell using lactate as an electron donor and the electrode as the electron acceptor. This property was shared by a large number of Fe(III)-reducing bacterial isolates. This is the first observation of a direct electrochemical reaction by an intact bacterial cell, which is believed to be possible due to the electron carrier(s) located at the cell surface involved in the reduction of the natural water insoluble electron acceptor, Fe(III).

• Journal of Microbiology and Biotechnology
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• v.4 no.4
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• pp.360-363
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• 1994

A colorimetric assay for $\gamma$-glutamyl transpeptidase ($\gamma$-CTP; E.C 2.3.2.2) employing 2, 4, 6-trinitrobenzene sulfonate (TNBS) to detect the amount of disappeared acceptor via transpeptidation, has been developed. Under the experimental conditions using L-$\gamma$-glutamyl ethyl ester and L-phenylalanine as $\gamma$-glutamyl donor and acceptor, respectively, it was found that the decreased absorbance of yellow color at 420 nm was strictly related to the amount of L-$\gamma$-glutamyl-L-phenylalanine (L-$\gamma$-Glu-L-Phe) formed, which was determined by DEAE-cellu-lose column chromatography. Concentrations of the enzyme and $\gamma$-glutamyl products were able to be determinedin the nanogram and nanomoles per milliliter range, respectively, with high precision and reliability. This novel assay system may therefore be a useful means for understanding of catalytic function of the $\gamma$-CTP spectrophotometrically without any usage of sophisticated instruments.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.104-110
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• 2012

Glycosynthase is an active site nucleophile mutant enzyme, prepared from glycosidase, which is capable of synthesizing oligosaccharide derivatives without the hydrolysis of the product. Thermoacidophilic ${\alpha}$-glucosidase of Thermoplasma acidophilum (AglA) exhibits a transglycosylating activity yielding various glycosides. AglA was converted to glycosynthase by the substitution of the catalytic nucleophile Asp-408 residue into non-nucleophile glycine in order to increase its ability to synthesize various glycosides by transglycosylation. The glycosynthase mutant was purified by Ni-NTA chromatography and its glycoside-synthesizing activity was measured by using an external nucleophile, sodium formate buffer, providing maltose as a donor and p-nitrophenyl-${\alpha}$-D-glucopyranoside ($pNP{\alpha}G$) as an acceptor, respectively. In addition, $pNP{\alpha}G$ was examined for its feasibility to act as both a donor and an acceptor, and products were compared with those of the wildtype enzyme. The mutant enzyme was found to catalyze the formation of a specific product from $pNP{\alpha}G$ with a yield of 42.5% without further hydrolysis, while the wild-type enzyme produced two $pNP{\alpha}G$ products at low yields. The results demonstrate the possibility of satisfactory yields for the reactions in the presence of small amounts of acceptor, and demonstrate that the high activity of the mutant, at pHs below neutrality, was applicable in the transfer of glucose from the natural donor.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.6
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• pp.646-651
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• 2018

This paper reports that the electronic properties at a $P3HT/TiO_2$ interface associated with exciton dissociation and transport can be tailored by the insertion of a graphene quantum dot (GQD) layer. For donor/acceptor interface modification in an $ITO/TiO_2/P3HT/Al$ photovoltaic (PV) device, a continuous GQD film was prepared by a sonication treatment in solution that simplifies the conventional processes, including laser fragmentation and hydrothermal treatment, which limits a variety of component layers and involves low cost processing. The high conductivity and favorable energy alignment for exciton dissociation of the GQD layer increased the fill factor and short circuit current. The origin of the improved parameters is discussed in terms of the broad light absorption and enhanced interfacial carrier transport.