Identification of “Hot Spots” to Improve Maturation of the moxSAASoti Fluorescent Protein at 37°C

N. K. Marynicha, *, and A. P. Savitskya, b

aBach Institute of Biochemistry, Federal Research Center “Fundamentals of Biotechnology”, Russian Academy of Sciences, Moscow, 119071 Russia

bDepartment of Chemistry, Moscow State University, Moscow, 119991 Russia

email: *marynich_n@mail.ru

Received 30 October, 2023

Abstract— A search for amino acid residues, whose replacement could contribute to the optimal maturation of the moxSAASoti fluorescent protein at 37°C, is carried out. For many other fluorescent proteins, this characteristic was improved by chance through many rounds of random mutagenesis; however, we managed to find two positions, 74 and 125, which clearly affect the process of moxSAASoti maturation, which is verified by introducing substitutions at these positions by site-directed and site-saturation mutagenesis.

Keywords: fluorescent proteins, site-saturation mutagenesis, maturation of fluorescent proteins

DOI: 10.3103/S0027131424700196