A new fluorogenic and chromogenic probe rhodamine B spirolactam (1) was developed that in aqueous solutions exhibited a highly selective and sensitive "turn-on" type spectral response toward NO detection following a NO-induced spiro-ring opening reaction in 1.
Menaquinone-7 (MK-7), a valuable vitamin K 2 , plays an important role in the prevention of osteoporosis and cardiovascular calcification. We chose B. subtilis 168 as the chassis for the modular metabolic engineering design to promote the biosynthesis of MK-7. The biosynthetic pathway of MK-7 was categorized into four modules, namely, the MK-7 pathway (Module I), the shikimate (SA) pathway (Module II), the methylerythritol phosphate (MEP) pathway (Module III), and the glycerol metabolism pathway (Module IV). Overexpression of menA (Module I) resulted in 6.6 ± 0.1 mg/L of MK-7 after 120 h fermentation, which was 2.1-fold that of the starting strain BS168NU (3.1 ± 0.2 mg/L). Overexpression of aroA, aroD, and aroE (Module II) had a negative effect on the synthesis of MK-7. Simultaneous overexpression of dxs, dxr, yacM, and yacN (Module III) enabled the yield of MK-7 to 12.0 ± 0.1 mg/L. Moreover, overexpression of glpD (Module IV) resulted in an increase of the yield of MK-7 to 13.7 ± 0.2 mg/L. Furthermore, deletion of dhbB reduced the consumption of the intermediate metabolite isochorismate, thus promoting the yield of MK-7 to 15.4 ± 0.6 mg/L. Taken together, the final resulting strain MK3-MEP123-Gly2-ΔdhbB with simultaneous overexpression of menA, dxs, dxr, yacM-yacN, glpD and deletion of dhbB enabled the yield of MK-7 to 69.5 ± 2.8 mg/L upon 144 h fermentation in a 2 L baffled flask.
Gold nanoparticle (AuNP) based colorimetric aptasensor have been developed for many analytes recently largely because of the ease of detection, high sensitivity, and potential for high-throughput analysis. Most of the target aptamers for detection have short sequences. However, the approach shows poor performance in terms of detection sensitivity for most of the long-sequence aptamers. To address this problem, for the first time, we split the 76 mer aptamer of 17β-estradiol into two short pieces to improve the AuNP based colorimetric sensitivity. Our results showed that the split P1 + P2 still retained the original 76 mer aptamer's affinity and specificity but increased the detection limit by 10-fold, demonstrating that as low as 0.1 ng/mL 17β-estradiol could be detected. The increased sensitivity may be caused by lower aptamer adsorption concentration and a lower affinity to the AuNPs of a short single-strand DNA (ssDNA) sequence. Our study provided a new way to use long-sequence aptamers to develop a highly sensitive AuNP-based colorimetric aptasensor.
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