title: Ð Ð°Ð·ÑÐ°Ð±Ð¾ÑÐºÐ° ÑÐ¸ÑÑÐµÐ¼ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Escherichia coli Ð¸ Pichia pastoris Ð´Ð»Ñ Ð¿ÐµÑÐµÑÐ°Ð±Ð¾ÑÐºÐ¸ Ð±Ð¸Ð¾ÑÐ°Ð·Ð»Ð°Ð³Ð°ÐµÐ¼ÑÑ Ð¾ÑÑ Ð¾Ð´Ð¾Ð² Ð¿Ð¸ÑÐµÐ²Ð¾Ð¹ Ð¿ÑÐ¾Ð¼ÑÑÐ»ÐµÐ½Ð½Ð¾ÑÑÐ¸
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Creation of expression systems for production of recombinant proteins in Escherichia coli and Pichia pastoris in order to resolve biodegradable pollutants of food industry
Creators: name: ÐÐ°ÐºÐ¾Ð²ÐµÐµÐ², ÐÐ¾Ð½ÑÑÐ°Ð½ÑÐ¸Ð½
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2024
Subjects: ÑÐ¸ÑÑÐµÐ¼Ñ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸
Ð¾ÑÑ Ð¾Ð´Ñ
Ð¿Ð¾Ð»Ð¸Ð¼ÐµÑÑ
Ð±Ð¸Ð¾Ð´ÐµÐ³ÑÐ°Ð´Ð°ÑÐ¸Ñ
ÐºÑÑÐ¸Ð½Ð°Ð·Ð°
expression systems
waste
polymers
biodegradation
cutinase

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° ÑÐ¾Ð·Ð´Ð°Ð½Ð¸Ñ ÑÑÐ°Ð¼Ð¼Ð¾Ð²-Ð¿ÑÐ¾Ð´ÑÑÐµÐ½ÑÐ¾Ð² ÑÐµÑÐ¼ÐµÐ½ÑÐ° ÐºÑÑÐ¸Ð½Ð°Ð·Ñ, ÑÐ¿Ð¾ÑÐ¾Ð±Ð½Ð¾Ð³Ð¾ ÑÑÐ¸Ð»Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°ÑÑ Ð±Ð¸Ð¾ÑÐ°Ð·Ð»Ð°Ð³Ð°ÐµÐ¼ÑÐµ Ð¾ÑÑÐ¾Ð´Ñ, Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ ÐºÑÐ»ÑÑÑÑÑ Ð¼ÐµÑÐ¸Ð»Ð¾ÑÑÐ¾ÑÐ½ÑÑ Ð´ÑÐ¾Ð¶Ð¶ÐµÐ¹ Pichia pastoris Ð¸ Ð¼Ð¸ÐºÑÐ¾Ð²Ð¾Ð´Ð¾ÑÐ¾ÑÐ»Ð¸ Chlamydomonas reinhardtii. Ð ÑÐ°Ð¼ÐºÐ°Ñ ÑÑÐ¾Ð¹ ÑÐµÐ»Ð¸ ÑÐµÑÐ°Ð»Ð¸ÑÑ ÑÐ»ÐµÐ´ÑÑÑÐ¸Ðµ Ð·Ð°Ð´Ð°ÑÐ¸: ÐÐ»Ð¾Ð½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ Ð¸ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ñ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÑ Ð³ÐµÐ½Ð¾Ð² ÐºÑÑÐ¸Ð½Ð°Ð· Ð² P. Ñastoris, Ð²ÑÑÐ²Ð»ÐµÐ½Ð¸Ðµ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÐ¸ Ñ ÑÐºÑÐ¿ÑÐµÑÑÐ¸ÐµÐ¹ Ð³ÐµÐ½Ð° ÑÐµÑÐ¼ÐµÐ½ÑÐ° Ñ Ð½Ð°Ð¸Ð±Ð¾Ð»ÑÑÐµÐ¹ ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑÑ Ð¸ ÐºÐ¾Ð½ÑÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ ÑÑÐ°Ð¼Ð¼Ð°-Ð¿ÑÐ¾Ð´ÑÑÐµÐ½ÑÐ° Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ ÐºÐ»ÐµÑÐ¾Ðº Ð´ÑÐ¾Ð¶Ð¶ÐµÐ¹. Ð¡Ð¾Ð·Ð´Ð°Ð½Ð¸Ðµ ÑÑÐ°Ð¼Ð¼Ð°-Ð¿ÑÐ¾Ð´ÑÑÐµÐ½ÑÐ° ÑÐ½Ð´Ð¾Ð½ÑÐºÐ»ÐµÐ°Ð·Ñ Cas9 Ð¸ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ ÑÐ¾Ð¾ÑÐ²ÐµÑÑÑÐ²ÑÑÑÐµÐ³Ð¾ Ð±ÐµÐ»ÐºÐ° Ð´Ð»Ñ ÐºÐ¾Ð½ÑÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ð¿ÑÐ¾Ð´ÑÑÐµÐ½ÑÐ° Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð¼Ð¸ÐºÑÐ¾Ð²Ð¾Ð´Ð¾ÑÐ¾ÑÐ»ÐµÐ¹ Chlamydomonas Ð ÐºÐ°ÑÐµÑÑÐ²Ðµ Ð¾Ð±ÑÐµÐºÑÐ° Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð²ÑÑÑÑÐ¿Ð°Ð» Ð¼Ð¸ÐºÑÐ¾Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð¼ Ð¸Ð· ÐºÐ¾Ð»Ð»ÐµÐºÑÐ¸Ð¸ Ð»Ð°Ð±Ð¾ÑÐ°ÑÐ¾ÑÐ¸Ð¸ Ð±Ð¸Ð¾ÑÐµÑÐ½Ð¾Ð»Ð¾Ð³Ð¸Ð¹ ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¾Ð³Ð¾ Ð¸Ð½ÑÑÐ¸ÑÑÑÐ° ÑÐ´ÐµÑÐ½Ð¾Ð¹ ÑÐ¸Ð·Ð¸ÐºÐ¸ Ð¸Ð¼. Ð.Ð. ÐÐ¾Ð½ÑÑÐ°Ð½ÑÐ¸Ð½Ð¾Ð²Ð°, ÐÐÐ¦ Â«ÐÑÑÑÐ°ÑÐ¾Ð²ÑÐºÐ¸Ð¹ Ð¸Ð½ÑÑÐ¸ÑÑÑÂ»: Aureobasidium pullulans ÐÐÐ 1116. ÐÑÐµÐ´Ð¼ÐµÑÐ¾Ð¼ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ ÑÐ²Ð»ÑÐµÑÑÑ Ð²ÑÑÐ²Ð»ÐµÐ½Ð¸Ðµ Ð³ÐµÐ½Ð¾Ð², ÐºÐ¾Ð´Ð¸ÑÑÑÑÐ¸Ñ ÐºÑÑÐ¸Ð½Ð°Ð·Ñ, Ð±ÐµÐ»ÐºÐ¾Ð², Ð¾Ð±Ð»Ð°Ð´Ð°ÑÑÐ¸Ñ Ð¿Ð¾Ð»Ð¸ÐºÐ°Ð¿ÑÐ¾Ð»Ð°ÐºÑÐ¾Ð½-Ð´ÐµÐ³ÑÐ°Ð´Ð¸ÑÑÑÑÐµÐ¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑÑ, Ð² ÐºÐ»ÐµÑÐºÐ°Ñ ÑÑÐ°Ð¼Ð¼Ð° A. pullulans ÐÐÐ 1116. Ð Ð°Ð±Ð¾ÑÐ° Ð²ÑÐ¿Ð¾Ð»Ð½ÐµÐ½Ð° Ð½Ð° Ð±Ð°Ð·Ðµ Ð»Ð°Ð±Ð¾ÑÐ°ÑÐ¾ÑÐ¸Ð¸ Ð±Ð¸Ð¾ÑÐµÑÐ½Ð¾Ð»Ð¾Ð³Ð¸Ð¸ ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¾Ð³Ð¾ Ð¸Ð½ÑÑÐ¸ÑÑÑÐ° ÑÐ´ÐµÑÐ½Ð¾Ð¹ ÑÐ¸Ð·Ð¸ÐºÐ¸ Ð¸Ð¼. Ð. Ð. ÐÐ¾Ð½ÑÑÐ°Ð½ÑÐ¸Ð½Ð¾Ð²Ð°, ÐÐÐ¦ Â«ÐÑÑÑÐ°ÑÐ¾Ð²ÑÐºÐ¸Ð¹ Ð¸Ð½ÑÑÐ¸ÑÑÑÂ» ÐÑÐ¸ Ð½Ð°Ð¿Ð¸ÑÐ°Ð½Ð¸Ðµ Ð»Ð¸ÑÐµÑÐ°ÑÑÑÐ½Ð¾Ð³Ð¾ Ð¾Ð±Ð·Ð¾ÑÐ° Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð»Ð¸ÑÑ ÑÐ»ÐµÐ´ÑÑÑÐ¸Ðµ Ð±Ð°Ð·Ñ Ð´Ð°Ð½Ð½ÑÑ Scopus, PubMed, Elibrary. Ð ÑÐ¾Ð´Ðµ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¸ Ð¿Ð¾Ð»ÑÑÐµÐ½Ñ: ÑÑÐ°Ð¼Ð¼-Ð¿ÑÐ¾Ð´ÑÑÐµÐ½Ñ ÐºÑÑÐ¸Ð½Ð°Ð·Ñ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð¼ÐµÑÐ¸Ð»Ð¾ÑÑÐ¾ÑÐ½ÑÑ Ð´ÑÐ¾Ð¶Ð¶ÐµÐ¹ Pichia pastoris, ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð° ÐºÑÑÐ¸Ð½Ð°Ð·Ð½Ð°Ñ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ñ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸ÐµÐ¼ ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ¾Ð³Ð¾ ÑÑÐ±ÑÑÑÐ°ÑÐ°; Ð¿ÑÐ¾Ð´ÑÑÐµÐ½Ñ ÑÐ½Ð´Ð¾Ð½ÑÐºÐ¾Ð»ÐµÐ°Ð·Ñ Cas 9 Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð±Ð°ÐºÑÐµÑÐ¸Ð°Ð»ÑÐ½Ð¾Ð¹ ÐºÑÐ»ÑÑÑÑÑ E. coli, Ð²ÑÐ´ÐµÐ»ÐµÐ½ ÑÐµÑÐ¼ÐµÐ½Ñ, ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð° ÐµÐ³Ð¾ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ.
description: Current work is devoted to development of strains producing enzyme cutinase, capable of utilizing biodegradable waste, based on the culture of methylotrophic yeast Pichia pastoris and microalgae Chlamydomonas reinhardtii.Â  Within this goal, the following objectives were pursued: Cloning and expression of potential cutinase genes in P. prastoris, identification of the sequence with expression of the enzyme gene with the highest enzymatic activity and construction of a yeast cell-based producer strain. Creation of a Cas9 endonuclease producer strain and isolation of the corresponding protein for the construction of a producer strain based on Chlamydomonas microalgae. The object of the study was a microorganism from the collection of the biotechnology laboratory of the St. Petersburg Institute of Nuclear Physics named after B.P. Konstantinov. Aureobasidium pullulans VKM 1116. The subject of the study is the identification of genes encoding cutinases, proteins possessing polycaprolactone-degrading activity, in cells of A. pullulans strain VKM 1116. The work was carried out at the Laboratory of Biotechnology of the St. Petersburg Institute of Nuclear Physics named after B. P. Konstantinov., SIC âKurchatov Instituteâ The following databases Scopus, PubMed, Elibrary were used in the literature review. In the course of work were obtained: a strain-producer of cutinase on the basis of methylotrophic yeast Pichia pastoris, the cutinase activity was established using a specific substrate; a producer of endonuclease Cas 9 on the basis of bacterial culture of E. coli., the enzyme was isolated and its activity was determined.