Nano-Porous Light-Emitting Silicon Chip as a Potential Biosensor Platform

Nano-porous silicon (PS) offers a potential platform for biosensors with benefits both in terms of light emission and the large functional surface area. A light emitting PS chip with a stable and functional surface was fabricated in our laboratory. When protein was deposited on it, the light emission was reduced in proportion to the protein concentration. Based on this property, we developed a rudimentary demonstration of a label-free sensor to detect bovine serum albumin (BSA). A serial concentration of BSA was applied to the light chip and the reduction in light emission was measured. The reduction of the light intensity was linearly related to the concentration of the BSA at concentrations below 10-5 M. The detection limit was 8×10-9 M

Biocatalytic Synthesis of a Novel Bioactive Ginsenoside Using UDP-Glycosyltransferase from Bacillus subtilis 168

Ginsenoside Rg3 is a bioactive compound from Panax ginseng and exhibits diverse notable biological properties. Glycosylation catalyzed by uridine diphosphate-dependent glycosyltransferase (UGT) is the final biosynthetic step of ginsenoside Rg3 and determines its diverse pharmacological activities. In the present study, promiscuous UGT Bs-YjiC from Bacillus subtilis 168 was expressed in Escherichia coli and purified via one-step nickel chelate affinity chromatography. The in vitro glycosylation reaction demonstrated Bs-Yjic could selectively glycosylate the C12 hydroxyl group of ginsenoside Rg3 to synthesize an unnatural ginsenoside Rd12. Ginsenoside Rd12 was about 40-fold more water-soluble than that of ginsenoside Rg3 (90 μM). Furthermore, in vitro cytotoxicity of ginsenoside Rd12 against diverse cancer cells was much stronger than that of ginsenoside Rg3. Our studies report the UGT-catalyzed synthesis of unnatural ginsenoside Rd12 for the first time. Ginsenoside Rd12 with antiproliferative activity might be further exploited as a potential anticancer drug

Biocatalytic Synthesis of Calycosin-7-O-β-D-Glucoside with Uridine Diphosphate–Glucose Regeneration System

Calycosin-7-O-β-D-glucoside (Cy7G) is one of the principal components of Radix astragali. This isoflavonoid glucoside is regarded as an indicator to assess the quality of R. astragali and exhibits diverse pharmacological activities. In this study, uridine diphosphate-dependent glucosyltransferase (UGT) UGT88E18 was isolated from Glycine max and expressed in Escherichia coli. Recombinant UGT88E18 could selectively and effectively glucosylate the C7 hydroxyl group of calycosin to synthesize Cy7G. A one-pot reaction by coupling UGT88E18 to sucrose synthase (SuSy) from G. max was developed. The UGT88E18–SuSy cascade reaction could recycle the costly uridine diphosphate glucose (UDPG) from cheap sucrose and catalytic amounts of uridine diphosphate (UDP). The important factors for UGT88E18–SuSy cascade reaction, including UGT88E18/SuSy ratios, different temperatures, and pH values, different concentrations of dimethyl sulfoxide (DMSO), UDP, sucrose, and calycosin, were optimized. We produced 10.5 g L−1 Cy7G in the optimal reaction conditions by the stepwise addition of calycosin. The molar conversion of calycosin was 97.5%, with a space–time yield of 747 mg L−1 h−1 and a UDPG recycle of 78 times. The present study provides a new avenue for the efficient and cost-effective semisynthesis of Cy7G and other valuable isoflavonoid glucosides by UGT–SuSy cascade reaction

Effect of protein immunogenicity and PEG size and branching on the anti-PEG immune response to PEGylated proteins

PEGylation has successfully improved the pharmacological properties of therapeutic proteins. However, polyethylene glycol (PEG) has been burdened by immunogenicity that renders a negative clinical effect on therapeutic proteins. The anti-PEG immune response to PEGylated proteins possibly depends on the nature of proteins and the conjugated methoxy PEG (mPEG). Thus, it is necessary to investigate the effects of protein immunogenicity, the extent of PEGylation, the molecular weight (Mw), and the branching of mPEG on the anti-PEG immune response. Ovalbumin, tetanus toxoid cm, TT-TT conjugate, and TT-bovine serum albumin conjugate were used as target proteins. PEGylated proteins with different extents of PEGylation were obtained by fractionation of the PEGylated IT with size exclusion chromatography. The PEGylated proteins with different Mw and branching of mPEG were obtained by modification of TT with linear mPEG (5 kDa and 20 kDa) and branched mPEG (20 kDa). The PEGylated proteins elicited high levels of anti-PEG antibodies (predominantly IgM and IgG1). The anti-PEG immune response depended on the immunogenicity of proteins, the extent of PEGylation, and the Mw of mPEG. In contrast, branching of mPEG had an insignificant effect on the anti-PEG immune response to the PEGylated proteins. (C) 2016 Elsevier Ltd. All rights reserved.</p

A label free electrochemical nanobiosensor study

Nano-porous silicon (PS) is an attractive material for incorporation into biosensors, because it has a large surface area combined with the ability to generate both optical and electrical signals. In this paper, we describe a label-free nanobiosensor for bovine serum albumin (BSA). Nano-porous silicon produced in our laboratory was functionalized prior to immobilization of anti-BSA antibody on the surface. Reaction with BSA in phosphate buffered saline (PBS) buffer resulted in an impedance change which was inversely proportional to the concentration of the analyte. The system PBS buffer/antigen-antibody/PS constitutes an electrolyte-insulator-semiconductor (EIS) structure, thus furnishing an impedance EIS nanobiosensor. The linear range of the sensor was 0-0.27 mg mL-1 and the sensitivity was less than 10 µg mL-1

LABEL-FREE ELECTROCHEMICAL DETECTION OF TETRACYCLINE BY AN APTAMER NANO-BIOSENSOR

A novel aptamer nano-porous silicon (PS) biosensor was investigated for the rapid determination of tetracyclines. Electrochemical impedance spectroscopy (EIS) was used to analyze the behavior of the sensor. The specific binding of tetracycline to the aptamer biosensor led to a decrease in impedance. The corresponding impedance spectra (Nyquist plots) were obtained when serial concentrations of tetracycline were added into the system. An equivalent electrical circuit was used to fit the impedance data. The linear range of the sensor was 2.1-62.4 nM

Isolation and Characterization of a Marine Microalga for Biofuel Production with Astaxanthin as a Co-Product

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