12 research outputs found
Facilities for macromolecular crystallography at the Helmholtz Zentrum Berlin
Three macromolecular crystallography MX beamlines at the Helmholtz Zentrum Berlin HZB are available for the regional, national and international structural biology user community. The state of the art synchrotron beamlines for MX BL14.1, BL14.2 and BL14.3 are located within the low section of the BESSY II electron storage ring. All beamlines are fed from a superconducting 7 T wavelength shifter insertion device. BL14.1 and BL14.2 are energy tunable in the range 5 16 keV, while BL14.3 is a fixed energy side station operated at 13.8 keV. All three beamlines are equipped with CCD detectors. BL14.1 and BL14.2 are in regular user operation providing about 200 beam days per year and about 600 user shifts to approximately 50 research groups across Europe. BL14.3 has initially been used as a test facility and was brought into regular user mode operation during the year 2010. BL14.1 has recently been upgraded with a microdiffractometer including a mini k goniometer and an automated sample changer. Additional user facilities include office space adjacent to the beamlines, a sample preparation laboratory, a biology laboratory safety level 1 and high end computing resources. In this article the instrumentation of the beamlines is described, and a summary of the experimental possibilities of the beamlines and the provided ancillary equipment for the user community is give
Pulp properties resulting from different pretreatments of wheat straw and their influence on enzymatic hydrolysis rate
Wheat straw was subjected to three different processes prior to saccharification, namely alkaline pulping, natural pulping and autohydrolysis, in order to study their effect on the rate of enzymatic hydrolysis. Parameters like medium concentration, temperature and time have been varied in order to optimize each method. Milling the raw material to a length of 4 mm beforehand showed the best
cost–value-ratio compared to other grinding methods studied. Before saccharification the pulp can be stored in dried form, leading to a high yield of glucose. Furthermore the relation of pulp properties (i.e. intrinsic viscosity, KLASON-lignin and hemicelluloses content, crystallinity, morphology) to cellulose hydrolysis is discussed
Workflow and Tools for Crystallographic Fragment Screening at the Helmholtz Zentrum Berlin
Fragment screening is a technique that helps to identify promising starting points for ligand design. Given that crystals of the target protein are available and display reproducibly high resolution X ray diffraction properties, crystallography is among the most preferred methods for fragment screening because of its sensitivity. Additionally, it is the only method providing detailed 3D information of the binding mode of the fragment, which is vital for subsequent rational compound evolution. The routine use of the method depends on the availability of suitable fragment libraries, dedicated means to handle large numbers of samples, state of the art synchrotron beamlines for fast diffraction measurements and largely automated solutions for the analysis of the results. Here, the complete practical workflow and the included tools on how to conduct crystallographic fragment screening CFS at the Helmholtz Zentrum Berlin HZB are presented. Preceding this workflow, crystal soaking conditions as well as data collection strategies are optimized for reproducible crystallographic experiments. Then, typically in a one to two day procedure, a 96 membered CFS focused library provided as dried ready to use plates is employed to soak 192 crystals, which are then flash cooled individually. The final diffraction experiments can be performed within one day at the robot mounting supported beamlines BL14.1 and BL14.2 at the BESSY II electron storage ring operated by the HZB in Berlin Adlershof Germany . Processing of the crystallographic data, refinement of the protein structures, and hit identification is fast and largely automated using specialized software pipelines on dedicated servers, requiring little user input. Using the CFS workflow at the HZB enables routine screening experiments. It increases the chances for successful identification of fragment hits as starting points to develop more potent binders, useful for pharmacological or biochemical application
Pulp properties resulting from different pretreatments of wheat straw and their influence on enzymatic hydrolysis rate
The macromolecular crystallography beamlines at BESSY II of the Helmholtz Zentrum Berlin Current status and perspectives
For a little over a decade now, the Macromolecular Crystallography MX group at the Helmholtz Zentrum Berlin HZB has been operating three state of the art synchrotron beamlines for MX at the BESSY II storage ring in Berlin. The three HZB MX beamlines, BL14.1, BL14.2 and BL14.3, serve a stable and growing user community of currently more than 100 independent research groups from Berlin, Germany and Europe. Every year, the beamlines provide close to 200 days of MX beamtime. Over time, the HZB MX beamlines and endstations, in particular BL14.1, have been continually developed and upgraded and, since 2010, they operate as the most productive MX beamlines in Germany. The environment of the beamlines includes various ancillary equipment as well as additional facilities, such as office space adjacent to the beamlines, a sample preparation laboratory, a safety level 1 biology laboratory HZB MX BioLab and all necessary computing resources. In this paper, the current status of the beamlines as well as the ongoing developments are describe