Chitosan-based hybrid immobilization in bienzymatic reactions and its application to the production of laminaribiose.

A hybrid-immobilization method was developed to improve the long-term stability of laminaribiose phosphorylase immobilized on epoxy supports Sepabeads EC-EP/S. Entrapment in chitosan retained all of the enzyme activity depending on the amount of entrapped solid materials and increased half-life by a factor of 10-94.4 h. No enzyme activity loss was determined during 12 times reuse. The immobilization method is also applicable to sucrose phosphorylase immobilized on Sepabeads EC-EP/S. Up to 31.9 g/L laminaribiose were produced in bienzymatic batch experiments with reaction-integrated product separation by adsorption on zeolites.

Development and characterization of a disposable plastic microarray printhead.

During the last decade microarrays have become a powerful analytical tool. Commonly microarrays are produced in a non-contact manner using silicone printheads. However, silicone printheads are expensive and not able to be used as a disposable. Here, we show the development and functional characterization of 8-channel plastic microarray printheads that overcome both disadvantages of their conventional silicone counterparts. A combination of injection-molding and laser processing allows us to produce a high quantity of cheap, customizable and disposable microarray printheads. The use of plastics (e.g., polystyrene) minimizes the need for surface modifications required previously for proper printing results. Time-consuming regeneration processes, cleaning procedures and contaminations caused by residual samples are avoided. The utilization of plastic printheads for viscous liquids, such as cell suspensions or whole blood, is possible. Furthermore, functional parts within the plastic printhead (e.g., particle filters) can be included. Our printhead is compatible with commercially available TopSpot devices but provides additional economic and technical benefits as compared to conventional TopSpot printheads, while fulfilling all requirements demanded on the latter. All in all, this work describes how the field of traditional microarray spotting can be extended significantly by low cost plastic printheads.

Studying the Impact of Wall Shear Stress on the Development and Performance of Electrochemically Active Biofilms.

A laminar flow reactor was designed that provides constant and reproducible growth conditions for the bioelectrochemical observation of electroactive bacteria (EAB). Experiments were performed using four reactors in parallel to enable the comparison of EAB growth behavior and bioelectrochemical performance under different hydrodynamic conditions while simultaneously keeping biological conditions identical. With regard to the moderate flow conditions found in wastewater treatment applications, the wall shear stress was adjusted to a range between 0.4 mPa to 2.9 mPa. Chronoamperometric data indicate that early stage current densities are improved by a moderate increase of the wall shear stress. In the same way, current onset times were increasing slightly towards higher values of the applied wall shear stress. Long-term observations of EAB performance showed a decrease in current density and a leveling of the trend observed for the early stages of biofilm growth.

Continuous enzymatic production and adsorption of laminaribiose in packed bed reactors.

Bienzymatic production of laminaribiose from sucrose and glucose was combined with adsorption on zeolite BEA to introduce a first capture and purification step. Downstream processing including washing and desorption steps was characterized and optimized on a milliliter scale in batch mode. Results were then transferred to a packed bed system for enzymatic production and adsorption where the influence of adsorbent particle diameter on purity and productivity was evaluated. Finally, a continuous enzymatic production of laminaribiose was conducted over 10 days. The subsequent downstream processing of the loaded zeolites led to purities of over 0.5 gLaminaribiose gsugar -1 in the desorbate with a total productivity of 5.6 mgLaminaribiose Lenzyme bed -1 h-1 without the use of recycles.

Immobilization and Characterization of E. gracilis Extract with Enriched Laminaribiose Phosphorylase Activity for Bienzymatic Production of Laminaribiose.

Immobilization methods and carriers were screened for immobilization of Euglena gracilis extract with laminaribiose phosphorylase activity. The extract was successfully immobilized on three different carriers via covalent linkage. Suitable immobilization carriers were Sepabeads EC-EP/S and ECR 8209M with epoxy groups and ECR 8309M with amino groups as functional units. Immobilization on Sepabeads EC-EP/S resulted in highest retained activity (65%). The immobilizates were characterized for pH, temperature, and buffer molarity preferences. The immobilized enzyme lost 48% of its activity when used seven times. Together with sucrose phosphorylase, laminaribiose phosphorylase was successfully applied for bienzymatic production of laminaribiose from sucrose and glucose with a final laminaribiose concentration of 14.3 ± 2.1 g/L (20% yield).

Expanding the elution by characteristic point method to columns with a finite number of theoretical plates.

The elution by characteristic point (ECP) method provides a rapid approach to determine whole isotherm data with small material usage. It is especially desired wherever the adsorbent or the adsorbate is expensive, toxic or only available in small amounts. However, the ECP method is limited to adsorbents that are well optimized for chromatographic use and therefore provide a high number of theoretical plates when packed into columns (2000 or more for Langmuir type isotherms are suggested). Here we present a novel approach that uses a new profile correction to apply the ECP method to poorly optimized adsorbents with less than 200 theoretical plates. Non-ideality effects are determined using a dead volume marker injection and the resulting marker profile is used to compensate the named effects considering their dependency from the actual concentration instead of assuming rectangular profiles. Experimental and literature data are used to compare the new ECP approach with batch method results.