Potential of Pichia pastoris for the production of industrial penicillin G acylase

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F17%3A00478086" target="_blank" >RIV/61388971:_____/17:00478086 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/17:10362915

Výsledek na webu

<a href="http://dx.doi.org/10.1007/s12223-017-0512-0" target="_blank" >http://dx.doi.org/10.1007/s12223-017-0512-0</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s12223-017-0512-0" target="_blank" >10.1007/s12223-017-0512-0</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Potential of Pichia pastoris for the production of industrial penicillin G acylase

Popis výsledku v původním jazyce

This study deals with the potential of Pichia pastoris X-33 for the production of penicillin G acylase (PGA(A)) from Achromobacter sp. CCM 4824. Synthetic gene matching the codon usage of P. pastoris was designed for intracellular and secretion-based production strategies and cloned into vectors pPICZ and pPICZ alpha under the control of AOX1 promoter. The simple method was developed to screen Pichia transformants with the intracellularly produced enzyme. The positive correlation between acylase production and pga gene dosage for both expression systems was demonstrated in small scale experiments. In fed-batch bioreactor cultures of X-33/PENS2, an extracellular expression system, total PGA(A) expressed from five copies reached 14,880 U/L of an active enzyme after 142 h, however, 60% of this amount retained in the cytosol. The maximum PGA(A) production of 31,000 U/L was achieved intracellularly from nine integrated gene copies of X-33/PINS2 after 90 h under methanol induction. The results indicate that in both expression systems the production level of PGA(A) is similar but there is a limitation in secretion efficiency.

Název v anglickém jazyce

Potential of Pichia pastoris for the production of industrial penicillin G acylase

Popis výsledku anglicky

This study deals with the potential of Pichia pastoris X-33 for the production of penicillin G acylase (PGA(A)) from Achromobacter sp. CCM 4824. Synthetic gene matching the codon usage of P. pastoris was designed for intracellular and secretion-based production strategies and cloned into vectors pPICZ and pPICZ alpha under the control of AOX1 promoter. The simple method was developed to screen Pichia transformants with the intracellularly produced enzyme. The positive correlation between acylase production and pga gene dosage for both expression systems was demonstrated in small scale experiments. In fed-batch bioreactor cultures of X-33/PENS2, an extracellular expression system, total PGA(A) expressed from five copies reached 14,880 U/L of an active enzyme after 142 h, however, 60% of this amount retained in the cytosol. The maximum PGA(A) production of 31,000 U/L was achieved intracellularly from nine integrated gene copies of X-33/PINS2 after 90 h under methanol induction. The results indicate that in both expression systems the production level of PGA(A) is similar but there is a limitation in secretion efficiency.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2017

Kód důvěrnosti údajů

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

Název periodika

Folia Microbiologica

ISSN

0015-5632

e-ISSN

—

Svazek periodika

62

Číslo periodika v rámci svazku

5

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

8

Strana od-do

417-424

Kód UT WoS článku

000409167900007

EID výsledku v databázi Scopus

2-s2.0-85014712491