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Article

Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions

1
Chiyoda Corporation, 3-13 Moriya-cho, Kanagawa-ku, Kanagawa, Yokohama 221-0022, Japan
2
Confocal Science Incorporated, 5-14-15 Fukasawa, Setagaya-ku, Tokyo 158-0081, Japan
3
Maruwa Foods and Biosciences Incorporated, 170-1 Tsutsui-cho, Yamatokoriyama, Nara 639-1123, Japan
*
Author to whom correspondence should be addressed.
Academic Editor: Borislav Angelov
Crystals 2021, 11(11), 1311; https://doi.org/10.3390/cryst11111311
Received: 30 September 2021 / Revised: 11 October 2021 / Accepted: 11 October 2021 / Published: 27 October 2021
(This article belongs to the Special Issue Protein Crystallography: Achievements and Challenges)
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. View Full-Text
Keywords: neutron diffraction; large crystal; reservoir control; real-time observation neutron diffraction; large crystal; reservoir control; real-time observation
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MDPI and ACS Style

Tanigawa, N.; Takahashi, S.; Yan, B.; Kamo, M.; Furubayashi, N.; Kubota, K.; Inaka, K.; Tanaka, H. Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions. Crystals 2021, 11, 1311. https://doi.org/10.3390/cryst11111311

AMA Style

Tanigawa N, Takahashi S, Yan B, Kamo M, Furubayashi N, Kubota K, Inaka K, Tanaka H. Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions. Crystals. 2021; 11(11):1311. https://doi.org/10.3390/cryst11111311

Chicago/Turabian Style

Tanigawa, Naoki, Sachiko Takahashi, Bin Yan, Masayuki Kamo, Naoki Furubayashi, Koji Kubota, Koji Inaka, and Hiroaki Tanaka. 2021. "Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions" Crystals 11, no. 11: 1311. https://doi.org/10.3390/cryst11111311

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