Creation of biocatalysts with prescribed properties

Abstract

Oxidoreductases are widely used in different branches of industry (particularly, in pharma ceutical) and analytical biotechnology. To optimize the use and to reduce production cost of a target product, the properties of a biocatalyst should be modified to meet requirements of a specific process or method of analysis. Problem of creation of biocatalysts with prescribed properties includes complex set of R&D such as an engineering of enzyme properties, con struction of recombinant strains overproducing the target protein, development of a large scale downstream process and preparation of a working form of a biocatalysts. In the present work, the basic features and practical examples of such an over all approach are considered for formate dehydrogenase and D amino acid oxidase.

The role of residues Arg169 and Arg220 in intersubunit interactions of yeast D-amino acid oxidase

“…An analo gous effect was observed in the case of the Cys108Phe, Cys108Ala, and Cys108Ser single mutations. 13 The first mutation leads to an increase in thermal stability of TvDAAO, whereas the second and third mutations cause a decrease in thermal stability.…”

“…The latter enzyme was obtained 12 in a highly purified state in 1976 and is most widely used in practice. [3][4][5] One can assume that, in view of the practical importance of this enzyme, numerous attempts were made in the last 30 years to crystallize TvDAAO. However, no structural data on this enzyme are available in the Protein Data Bank (PDB) and there are no publications concern ing this problem, which suggests that all the attempts were unsuccessful.…”

“…We have also carried out a large scale screen ing of the crystallization conditions for wild type TvDAAO but failed to obtain crystals. Only after obtaining the Cys108Phe mutant of TvDAAO, 13 we succeeded in grow ing crystals of this enzyme and then solved its structure first at 2.8 Å resolution 14 and then at 1.8 Å resolution. 2 Porcine kidney D amino acid oxidase in vivo is a mono mer, whereas the enzymes from other sources exist in na ture as dimers.…”

Soils of Chernevaya Taiga of Western Siberia — Morphology, Agrochemical Features, Microbiome

“…Почти все эти леса подстилочные, с преобладанием зеленых мхов, мелкотравных и кустарничковых видов. Среди них лишь бореально-неморальновысокотравные леса рассматриваются некоторыми авторами как относительно полно отвечающие заключительному этапу аутогенной сукцессии (24).…”