Search

Your search keyword '"Honda, Kohsuke"' showing total 218 results
Start Over Author "Honda, Kohsuke" Search Limiters Available in Library Collection
218 results on '"Honda, Kohsuke"'

13. Chitin- and Streptavidin- Mediated Affinity Purification Systems: A Screening Platform for Enzyme Discovery

Author

1000060909125, Kato, Shunsuke, Takeuchi, Koki, Iwaki, Motonao, 1000060344123, 0000-0002-4349-566X, Miyazaki, Kentaro, 1000090403162, 0000-0001-9069-7574, Honda, Kohsuke, 1000020222226, 0000-0002-2215-935X, Hayashi, Takashi, 1000060909125, Kato, Shunsuke, Takeuchi, Koki, Iwaki, Motonao, 1000060344123, 0000-0002-4349-566X, Miyazaki, Kentaro, 1000090403162, 0000-0001-9069-7574, Honda, Kohsuke, 1000020222226, 0000-0002-2215-935X, and Hayashi, Takashi

Abstract

This is the peer reviewed version of the following article: Kato Shunsuke, Takeuchi Koki, Iwaki Motonao, et al., "Chitin- and Streptavidin- Mediated Affinity Purification Systems: A Screening Platform for Enzyme Discovery", Angewandte Chemie International Edition, 26, June, 2023, which has been published in final form at https://doi.org/10.1002/anie.202303764. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited., Affinity purification of recombinant proteins is an essential technique in biotechnology. However, current affinity purification methods are very cost-intensive, and this imposes limits on versatile use of affinity purification for obtaining purified proteins for a variety of applications. To overcome this problem, we developed a new affinity purification system which we call CSAP (chitin- and streptavidin-mediated affinity purification) for low-cost purification of Strep-tag II fusion proteins. The CSAP system is designed to utilize commercially available chitin powder as a chromatography matrix, thereby significantly improving the cost-efficiency of protein affinity purification. We investigated the CSAP system for protein screening in 96-well format as a demonstration. Through the screening of 96 types of purified hemoproteins, several proteins capable of the catalytic diastereodivergent synthesis of cyclopropanes were identified as candidates for an abiotic carbene transfer reaction.

Published
2023

35. Synthetic metabolic engineering-a novel, simple technology for designing a chimeric metabolic pathway

Author

Ye Xiaoting, Honda Kohsuke, Sakai Takaaki, Okano Kenji, Omasa Takeshi, Hirota Ryuichi, Kuroda Akio, and Ohtake Hisao

Subjects
Microbiology, QR1-502
Abstract

Abstract Background The integration of biotechnology into chemical manufacturing has been recognized as a key technology to build a sustainable society. However, the practical applications of biocatalytic chemical conversions are often restricted due to their complexities involving the unpredictability of product yield and the troublesome controls in fermentation processes. One of the possible strategies to overcome these limitations is to eliminate the use of living microorganisms and to use only enzymes involved in the metabolic pathway. Use of recombinant mesophiles producing thermophilic enzymes at high temperature results in denaturation of indigenous proteins and elimination of undesired side reactions; consequently, highly selective and stable biocatalytic modules can be readily prepared. By rationally combining those modules together, artificial synthetic pathways specialized for chemical manufacturing could be designed and constructed. Results A chimeric Embden-Meyerhof (EM) pathway with balanced consumption and regeneration of ATP and ADP was constructed by using nine recombinant E. coli strains overproducing either one of the seven glycolytic enzymes of Thermus thermophilus, the cofactor-independent phosphoglycerate mutase of Pyrococcus horikoshii, or the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase of Thermococcus kodakarensis. By coupling this pathway with the Thermus malate/lactate dehydrogenase, a stoichiometric amount of lactate was produced from glucose with an overall ATP turnover number of 31. Conclusions In this study, a novel and simple technology for flexible design of a bespoke metabolic pathway was developed. The concept has been testified via a non-ATP-forming chimeric EM pathway. We designated this technology as “synthetic metabolic engineering”. Our technology is, in principle, applicable to all thermophilic enzymes as long as they can be functionally expressed in the host, and thus would be potentially applicable to the biocatalytic manufacture of any chemicals or materials on demand.

Published
2012
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results