Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P-pastoris: functional characterisation of a novel promoter

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F17%3A43914615" target="_blank" >RIV/60461373:22330/17:43914615 - isvavai.cz</a>

Result on the web

<a href="https://link.springer.com/article/10.1007%2Fs00253-017-8123-x" target="_blank" >https://link.springer.com/article/10.1007%2Fs00253-017-8123-x</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s00253-017-8123-x" target="_blank" >10.1007/s00253-017-8123-x</a>

Result language

angličtina

Original language name

Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P-pastoris: functional characterisation of a novel promoter

Original language description

As Pichia pastoris (syn. Komagataella sp.) yeast can secrete pure recombinant proteins at high rates, it is a desirable production system. The function of a novel synthetic variant of the AOX1 promoter was characterised comprehensively using a strain secreting Candida antarctica lipase B (CALB) as a model. A new time-saving approach was introduced to determine, in only one experiment, the hitherto unknown relationship between specific product formation rate (q (p)) and specific growth rate (mu). Tight control of recombinant protein formation was possible in the absence of methanol, while using glycerol as a sole carbon/energy source. CALB was not synthesised during batch cultivation in excess glycerol (&gt; 10 g l(-1)) and at a growth rate close to mu (max) (0.15 h(-1)). Between 0.017 and 0.115 h(-1) in glycerol-limited fedbatch cultures, basal levels of q (p) &gt; 0.4 mg g(-1) h(-1) CALB were reached, independent of the mu at which the culture grew. At mu &gt; 0.04 h(-1), an elevated q (p) occurred temporarily during the first 20 h after changing to fedbatch mode and decreased thereafter to basal. In order to accelerate the determination of the q (p)(mu) relationship (kinetics of product formation), the entire mu range was covered in a single fedbatch experiment. By linearly increasing and decreasing glycerol addition rates, mu values were repeatedly shifted from 0.004 to 0.074 h(-1) and vice versa. Changes in q (p) were related to changes in mu. A rough estimation of mu range suitable for production was possible in a single fedbatch, thus significantly reducing the experimental input over previous approaches comprising several experiments.

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

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Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2017

Confidentiality

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

Name of the periodical

Applied Microbiology and Biotechnology

ISSN

0175-7598

e-ISSN

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Volume of the periodical

101

Issue of the periodical within the volume

8

Country of publishing house

US - UNITED STATES

Number of pages

14

Pages from-to

3163-3176

UT code for WoS article

000399167800012

EID of the result in the Scopus database

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