Recombinant Strains Producing Thermomyces lanuginosus Thermostable Lipase and their Use in Heterogeneous Biocatalysis, Including Processes of Low-Temperature Synthesis of Esters Full article
| Journal |
Applied Biochemistry and Microbiology
ISSN: 0003-6838 , E-ISSN: 1608-3024 |
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| Output data | Year: 2022, Volume: 58, Number: 8, Pages: 887–898 Pages count : 12 DOI: 10.1134/S0003683822080026 | ||||
| Tags | Escherichia coli, Pichia pastoris, recombinant producer strains, Thermomyces lanuginosus lipase gene, immobilization, biocatalysts, esterification | ||||
| Authors |
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| Affiliations |
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Funding (1)
| 1 | Ministry of Science and Higher Education of the Russian Federation | 0239-2021-0005 |
Abstract:
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.
Cite:
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 AN OpenAlex
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 AN OpenAlex
Original:
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
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
Dates:
| Submitted: | Jul 6, 2021 |
| Accepted: | Sep 9, 2021 |
| Published print: | Dec 1, 2022 |
| Published online: | Dec 16, 2022 |
Identifiers:
| Web of science: | WOS:000899987600001 |
| Scopus: | 2-s2.0-85144198053 |
| Elibrary: | 53763040 |
| Chemical Abstracts: | 2022:3131543 |
| OpenAlex: | W4311897066 |