Gram-scale Production of Recombinant Microbial Enzymes in Shake Flasks

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F18%3A00100791" target="_blank" >RIV/00216224:14310/18:00100791 - isvavai.cz</a>

Alternative codes found

RIV/00159816:_____/18:00068669

Result on the web

<a href="http://dx.doi.org/10.1093/femsle/fnx265" target="_blank" >http://dx.doi.org/10.1093/femsle/fnx265</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1093/femsle/fnx265" target="_blank" >10.1093/femsle/fnx265</a>

Result language

angličtina

Original language name

Gram-scale Production of Recombinant Microbial Enzymes in Shake Flasks

Original language description

Heterologous production of recombinant proteins is a cornerstone of microbiological and biochemical research as well as various biotechnological processes. Yields and quality of produced proteins have a tremendous impact on structural and enzymology studies, development of new biopharmaceuticals and establishing new biocatalytic processes. Majority of current protocols for recombinant protein expression in Escherichia coli exploit batch cultures with complex media, often providing low yields of the target protein due to oxygen transfer limitation, rapid depletion of carbon sources, and pH changes during the cultivation. Recently introduced EnBase technology enables fed-batch-like cultivations in shake flasks with continuous glucose release from a soluble starch. In this study, we critically compare the yields of fourteen model enzymes in E. coli cultured in a novel semi-defined medium and in a complex medium. Significant improvements in the volumetric yields 2-31 times were observed for all tested enzymes expressed in enzymatic fed-batch-like cultures with no adverse impact on enzyme structure, stability, or activity. Exceptional yields, higher than one gram of protein per litre of culture, were obtained with six enzymes. We conclude that the novel semi-defined medium tested in this study provides a robust improvement of protein yields in shake flasks without investment into costly bioreactors.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10606 - Microbiology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2018

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

FEMS Microbiology Letters

ISSN

0378-1097

e-ISSN

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Volume of the periodical

365

Issue of the periodical within the volume

2

Country of publishing house

GB - UNITED KINGDOM

Number of pages

22

Pages from-to

1-22

UT code for WoS article

000429310300008

EID of the result in the Scopus database

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