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Keywords
The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates.
Murib, M. S. ; Yeap, W. S. ; Martens, D. ; Liu, X. ; Bienstman, P. ; Fahlman, M. ; Schöning, Michael Josef ; Michiels, L. ; Haenen, K. ; Serpengüzel, A. ; Wagner, Patrick
In this study we show an optical biosensor concept, based on elastic light scattering from sapphire microspheres. Transmitted and elastic scattering intensity of the microspheres (radius 500 μm, refractive index 1.77) on an optical fiber half coupler is analyzed at 1510 nm. The 0.43 nm angular mode spacing of the resonances is comparable to the angular mode spacing value estimated using the optical size of the microsphere. The spectral linewidths of the resonances are in the order of 0.01 nm, which corresponds to quality factors of approximately 105. A polydopamine layer is used as a functionalizing agent on sapphire microspherical resonators in view of biosensor implementation. The varying layer thickness on the microsphere is determined as a function of the resonance wavelength shift. It is shown that polymer functionalization has a minor effect on the quality factor. This is a promising step toward the development of an optical biosensor.
Mikulics, M. ; Marso, M. ; Cámara Mayorga, I. ; Gusten, R. ; Stancek, S. ; Michael, E. A. ; Schieder, R. ; Wolter, M. ; Buca, D. ; Förster, Arnold ; Kordos, P. ; Lüth, H.
Ermelenko, Y. ; Yoshinobu, T ; Mourzina, Y. ; Levichev, S. ; Furuichi, K. ; Vlasov, Y. ; Schöning, Michael Josef ; Iwasaki, H.
Miyamoto, Ko-ichiro ; Wagner, Torsten ; Yoshinobu, Tatsuo ; Kanoh, Shin`ichiro ; Schöning, Michael Josef
Miyamoto, Ko-ichiro ; Wagner, Torsten ; Yoshinobu, Tatsuo ; Kanoh, Shin`ichiro ; Schöning, Michael Josef
Buniatyan, Vahe V. ; Abouzar, Maryam H. ; Martirosyan, Norayr W. ; Schubert, Jürgen ; Gevorgian, Spartak ; Schöning, Michael Josef ; Poghossian, Arshak
Christiaens, P. ; Abouzar, Maryam H. ; Poghossian, Arshak ; Wagner, Torsten ; Bijnens, N. ; Williams, O. A. ; Daenen, M. ; Haenen, K. ; Douthéret, O. ; Haen, J. d´ ; Mekhalif, Z. ; Schöning, Michael Josef ; Wagner, P.
Abouzar, Maryam H. ; Poghossian, Arshak ; Razavi, Arash ; Besmehn, Astrid ; Bijnens, Nathalie ; Williams, Oliver A. ; Haenen, Ken ; Wagner, Patrick ; Schöning, Michael Josef
The presentation of enzymes on viral scaffolds has beneficial effects such as an increased enzyme loading and a prolonged reusability in comparison to conventional immobilization platforms. Here, we used modified tobacco mosaic virus (TMV) nanorods as enzyme carriers in penicillin G detection for the first time. Penicillinase enzymes were conjugated with streptavidin and coupled to TMV rods by use of a bifunctional biotin-linker. Penicillinase-decorated TMV particles were characterized extensively in halochromic dye-based biosensing. Acidometric analyte detection was performed with bromcresol purple as pH indicator and spectrophotometry. The TMV-assisted sensors exhibited increased enzyme loading and strongly improved reusability, and higher analysis rates compared to layouts without viral adapters. They extended the half-life of the sensors from 4 - 6 days to 5 weeks and thus allowed an at least 8-fold longer use of the sensors. Using a commercial budget-priced penicillinase preparation, a detection limit of 100 µM penicillin was obtained. Initial experiments also indicate that the system may be transferred to label-free detection layouts.
Penicillin detection by means of field-effect based sensors: EnFET, capacitive EIS sensor or LAPS?
(2001)
Poghossian, Arshak ; Yoshinobu, Tatsuo ; Simonis, A. ; Ecken, H. ; Lüth, Hans ; Schöning, Michael Josef
Penicillin detection by means of field-effect based sensors: EnFET, capacitive EIS sensor or LAPS?
(2000)
Siqueira, José R. Jr. ; Abouzar, Maryam H. ; Poghossian, Arshak ; Zucolotto, Valtencir ; Oliveira, Osvaldo N. Jr. ; Schöning, Michael Josef
Gun, Jenny ; Rizkov, Dan ; Lev, Ovadia ; Abouzar, Maryam H. ; Poghossian, Arshak ; Schöning, Michael Josef
Optoelectronic Properties of Nanostructured Ensembles Controlled by Biomolecular Logic Systems
(2008)
Pita, Marcos ; Krämer, Melina ; Zouh, Jian ; Poghossian, Arshak ; Schöning, Michael Josef ; Fernandez, Victor M. ; Katz, Evgeny
Optimization of the immobilization of bacterial spores on glass substrates with organosilanes
(2016)
Arreola, Julio ; Mätzkow, Malte ; Durán, Marlena Palomar ; Greeff, Anton ; Keusgen, Michael ; Schöning, Michael Josef
Spores can be immobilized on biosensors to function as sensitive recognition elements. However, the immobilization can affect the sensitivity and reproducibility of the sensor signal. In this work, three different immobilization strategies with organosilanes were optimized and characterized to immobilize Bacillus atrophaeus spores on glass substrates. Five different silanization parameters were investigated: nature of the solvent, concentration of the silane, silanization time, curing process, and silanization temperature. The resulting silane layers were resistant to a buffer solution (e.g., Ringer solution) with a polysorbate (e.g., Tween®80) and sonication.