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Novel device and strategy for growing large, high-quality protein crystals by controlling crystallization conditions

DOI: 10.3390/cryst11111311 DOI Help

Authors: Naoki Tanigawa (Chiyoda Corporation) , Sachiko Takahashi (Confocal Science Incorporated) , Bin Yan (Confocal Science Incorporated) , Masayuki Kamo (Maruwa Foods and Biosciences Incorporated) , Naoki Furubayashi (Maruwa Foods and Biosciences Incorporated) , Koji Kubota (Chiyoda Corporation) , Koji Inaka (Maruwa Foods and Biosciences Incorporated) , Hiroaki Tanaka (Confocal Science Incorporated)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Crystals , VOL 11

State: Published (Approved)
Published: November 2021
Diamond Proposal Number(s): 12421

Open Access Open Access

Abstract: Neutron diffraction experiments are informative for determining the locations of hydrogen atoms in protein molecules; however, much larger crystals are needed than those required for X-ray diffraction. Thus, additional techniques are required to grow larger crystals. Here, a unique crystallization device and strategy for growing large protein crystals are introduced. The device uses two micropumps to control crystal growth by altering the precipitant concentration and regulating the pinpoint injection of dry air flow to the crystallization cell. Furthermore, the crystal growth can be observed in real time. Preliminary microbatch crystallization experiments at various concentration ranges of polyethylene glycol (PEG) 4000 and sodium chloride were first performed to elucidate optimized crystallization conditions. Based on these results, a device to precisely control the sodium chloride and PEG concentrations and the supply of dry air to the crystallization cell was used, and 1.8 mm lysozyme and 1.5 mm alpha-amylase crystals with good reproducibility were obtained. X-ray data sets of both crystals were collected at room temperature at BL2S1 of the Aichi Synchrotron Radiation Center and confirmed that these crystals were of high quality. Therefore, this crystallization device and strategy were effective for growing large, high-quality protein crystals.

Journal Keywords: neutron diffraction; large crystal; reservoir control; real-time observation

Subject Areas: Biology and Bio-materials, Technique Development


Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)

Other Facilities: BL2S1 at Aichi Science & Technology Foundation

Added On: 03/11/2021 09:39

Documents:
crystals-11-01311-v2.pdf

Discipline Tags:

Technique Development - Life Sciences & Biotech Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)