Gram-scale Production of Recombinant Microbial Enzymes in Shake Flasks

Kód výsledku v 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>

Nalezeny alternativní kódy

RIV/00159816:_____/18:00068669

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

Gram-scale Production of Recombinant Microbial Enzymes in Shake Flasks

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

Gram-scale Production of Recombinant Microbial Enzymes in Shake Flasks

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

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

Rok uplatnění

2018

Kód důvěrnosti údajů

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

Název periodika

FEMS Microbiology Letters

ISSN

0378-1097

e-ISSN

—

Svazek periodika

365

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

22

Strana od-do

1-22

Kód UT WoS článku

000429310300008

EID výsledku v databázi Scopus

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