TY - JOUR A1 - Reinicke, Stefan A1 - Fischer, Thilo A1 - Bramski, Julia A1 - Pietruszka, Jörg A1 - Böker, Alexander T1 - Biocatalytically active microgels by precipitation polymerization of N-isopropyl acrylamide in the presence of an enzyme JF - RSC Advances N2 - We present a novel protocol for the synthesis of enzymatically active microgels. The protocol is based on the precipitation polymerization of N-isopropylacrylamide (NIPAm) in the presence of an enzyme and a protein binding comonomer. A basic investigation on the influence of different reaction parameters such as monomer concentration and reaction temperature on the microgel size and size distribution is performed and immobilization yields are determined. Microgels exhibiting hydrodynamic diameters between 100 nm and 1 mu m and narrow size distribution could be synthesized while about 31-44% of the enzyme present in the initial reaction mixture can be immobilized. Successful immobilization including a verification of enzymatic activity of the microgels is achieved for glucose oxidase (GOx) and 2-deoxy-d-ribose-5-phosphate aldolase (DERA). The thermoresponsive properties of the microgels are assessed and discussed in the light of activity evolution with temperature. The positive correlation of enzymatic activity with temperature for the GOx containing microgel originates from a direct interaction of the enzyme with the PNIPAm based polymer matrix whose magnitude is highly influenced by temperature. Y1 - 2019 U6 - https://doi.org/10.1039/c9ra04000e SN - 2046-2069 VL - 9 IS - 49 SP - 28377 EP - 28386 PB - Royal Society of Chemistry CY - Cambridge ER - TY - JOUR A1 - Zhang, Shuhao A1 - Bramski, Julia A1 - Tutus, Murat A1 - Pietruszka, Jörg A1 - Böker, Alexander A1 - Reinicke, Stefan T1 - A Biocatalytically Active Membrane Obtained from Immobilization of 2-Deoxy-D-ribose-5-phosphate Aldolase on a Porous Support JF - ACS applied materials & interfaces N2 - Aldol reactions play an important role in organic synthesis, as they belong to the class of highly beneficial C-C-linking reactions. Aldol-type reactions can be efficiently and stereoselectively catalyzed by the enzyme 2-deoxy-D-ribose-5-phosphate aldolase (DERA) to gain key intermediates for pharmaceuticals such as atorvastatin. The immobilization of DERA would open the opportunity for a continuous operation mode which gives access to an efficient, large-scale production of respective organic intermediates. In this contribution, we synthesize and utilize DERA/polymer conjugates for the generation and fixation of a DERA bearing thin film on a polymeric membrane support. The conjugation strongly increases the tolerance of the enzyme toward the industrial relevant substrate acetaldehyde while UV-cross-linkable groups along the conjugated polymer chains provide the opportunity for covalent binding to the support. First, we provide a thorough characterization of the conjugates followed by immobilization tests on representative, nonporous cycloolefinic copolymer supports. Finally, immobilization on the target supports constituted of polyacrylonitrile (PAN) membranes is performed, and the resulting enzymatically active membranes are implemented in a simple membrane module setup for the first assessment of biocatalytic performance in the continuous operation mode using the combination hexanal/acetaldehyde as the substrate. KW - 2-deoxy-D-ribose-5-phoshphate aldolase KW - enzyme immobilization KW - enzymatically active membrane KW - enzyme/polymer conjugate KW - self-assembly Y1 - 2019 U6 - https://doi.org/10.1021/acsami.9b12029 SN - 1944-8244 SN - 1944-8252 VL - 11 IS - 37 SP - 34441 EP - 34453 PB - American Chemical Society CY - Washington ER -