Recombinant Strains Producing Thermomyces lanuginosus Thermostable Lipase and their Use in Heterogeneous Biocatalysis, Including Processes of Low-Temperature Synthesis of Esters | Sciact - CRIS-система Института катализа

Recombinant Strains Producing Thermomyces lanuginosus Thermostable Lipase and their Use in Heterogeneous Biocatalysis, Including Processes of Low-Temperature Synthesis of Esters Научная публикация

Журнал

Applied Biochemistry and Microbiology
ISSN: 0003-6838 , E-ISSN: 1608-3024

Вых. Данные

Год: 2022, Том: 58, Номер: 8, Страницы: 887–898 Страниц : 12 DOI: 10.1134/S0003683822080026

Ключевые слова

Escherichia coli, Pichia pastoris, recombinant producer strains, Thermomyces lanuginosus lipase gene, immobilization, biocatalysts, esterification

Авторы

Beklemishev A.B. ^1,2 , Pykhtina M.B. ¹ , Perminova L.V. ² , Kovalenko G.A. ²

Организации

1	Research Institute of Biochemistry, Federal Research Center for Fundamental and Translational Medicine, Novosibirsk, 630117 Russia
2	Boreskov Institute of Catalysis, Russian Academy of Sciences, Siberian Branch, Novosibirsk, 630090 Russia

Информация о финансировании (1)

Министерство науки и высшего образования Российской Федерации (с 15 мая 2018)

0239-2021-0005

This work was devoted to the construction of recombinant strains Escherichia coli BL21 (DE3) and Pichia pastoris X33, which produce a 1,3-specific thermostable lipase from Thermomyces lanuginosus. The sequences of two lipase genes were optimized for expression in bacteria and methylotrophic yeasts, then synthesized and cloned in the corresponding expression vectors. As a result of genetic engineering manipulations, E. coli and P. pastoris strains were constructed that efficiently produced T. lanuginosus recombinant lipase. Recombinant E. coli clones accumulated lipase in the cytoplasm at a level of 30–40% of the total cellular protein. Recombinant P. pastoris clones secreted lipase into the culture medium at a concentration of at least 1 g/L. Lipases produced by the recombinant clones, designated as rE.coli/lip and rPichia/lip, contained a six-histidine sequence (-His6) in the C-terminal region. The resulting lipases were immobilized on/in solid inorganic supports in order to develop heterogeneous biocatalysts (HBs) for the enzymatic conversion of triglycerides and fatty acids. The rPichia/lip enzyme was adsorbed on mesoporous silica and macroporous carbon aerogel. The properties of the prepared HBs, their enzymatic activity, substrate specificity, and operational stability were studied in the reaction of esterification of fatty acids with aliphatic alcohols in organic solvents at 20 ± 2°C. It was found that immobilized lipases had a relatively wide substrate specificity, as well as high operational stability, and the prepared HBs almost completely retained their high esterifying activity for several tens of reaction cycles.

Beklemishev A.B. , Pykhtina M.B. , Perminova L.V. , Kovalenko G.A.
Recombinant Strains Producing Thermomyces lanuginosus Thermostable Lipase and their Use in Heterogeneous Biocatalysis, Including Processes of Low-Temperature Synthesis of Esters
Applied Biochemistry and Microbiology. 2022. V.58. N8. P.887–898. DOI: 10.1134/S0003683822080026 WOS Scopus РИНЦ CAPlus OpenAlex

Беклемишев А.Б. , Пыхтина М.Б. , Перминова Л.В. , Коваленко Г.А.
Рекомбинантные штаммы-продуценты термостабильной липазы из Thermomyces lanuginosus и их применение в гетерогенном биокатализе, в том числе, в процессах низкотемпературного синтеза сложных эфиров
Биотехнология. 2021. Т.37. №5. С.5-19. DOI: 10.21519/0234-2758-2021-37-5-5-19 Scopus РИНЦ OpenAlex

Поступила в редакцию:	6 июл. 2021 г.
Принята к публикации:	9 сент. 2021 г.
Опубликована в печати:	1 дек. 2022 г.
Опубликована online:	16 дек. 2022 г.

Web of science:	WOS:000899987600001
Scopus:	2-s2.0-85144198053
РИНЦ:	53763040
Chemical Abstracts:	2022:3131543
OpenAlex:	W4311897066

БД	Цитирований
Scopus	3
Web of science	3
РИНЦ	3
OpenAlex	5