In-silico driven engineering of enantioselectivity of a penicillin G acylase towards active pharmaceutical ingredients

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F16%3A10368393" target="_blank" >RIV/00216208:11310/16:10368393 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388971:_____/16:00483930

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.molcatb.2016.11.014" target="_blank" >http://dx.doi.org/10.1016/j.molcatb.2016.11.014</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.molcatb.2016.11.014" target="_blank" >10.1016/j.molcatb.2016.11.014</a>

Jazyk výsledku

angličtina

Název v původním jazyce

In-silico driven engineering of enantioselectivity of a penicillin G acylase towards active pharmaceutical ingredients

Popis výsledku v původním jazyce

Penicillin G acylase is one of the most employed enzymes in the pharmaceutical industry due to its role in the biotransformation of semi-synthetic Beta-lactam antibiotics. Recently, the enantioselectivity of the penicillin G acylase markedly broadened its application potential. In this study, we have evaluated effects of in-silico replacements of acyl-binding subsite residue Phe24Beta of the enzyme from Achromobacter sp. CCM 4824 to seven markedly smaller amino acids on its enantioselectivity towards industrially relevant compounds. Models of the two most promising mutants bearing substitutions PheBeta24Ala and PheBeta24Cys were investigated using molecular docking calculations. The Cys substitution revealed much better enantioselectivity traits with a set of seven substrates. To verify the relevance of in-silico predictions, we constructed a PGA. A + Phe24BetaCys mutant and determined its enantioselectivity in biocatalytic reactions. Since we experimentally confirmed all these predictions, we expanded our in-silico analysis to another set of seven compounds: the prediction suggested increased enantioselectivity for N-phenylacetyl-p-F-α-phenylalanine. The (R)-enantiomer of this substrate is used as a building block in synthesis of important anti-cancer agent Abarelix. The enantioselectivity of PGA. A + Phe24BetaCys mutant towards this substrate was improved by 75percent reaching E-value of about 70. Our results suggest the rapid identification of interesting replacements altering enantioselectivity using in-silico approach as the way for further expanding biotechnological application of penicillin G acylase.

Název v anglickém jazyce

In-silico driven engineering of enantioselectivity of a penicillin G acylase towards active pharmaceutical ingredients

Popis výsledku anglicky

Penicillin G acylase is one of the most employed enzymes in the pharmaceutical industry due to its role in the biotransformation of semi-synthetic Beta-lactam antibiotics. Recently, the enantioselectivity of the penicillin G acylase markedly broadened its application potential. In this study, we have evaluated effects of in-silico replacements of acyl-binding subsite residue Phe24Beta of the enzyme from Achromobacter sp. CCM 4824 to seven markedly smaller amino acids on its enantioselectivity towards industrially relevant compounds. Models of the two most promising mutants bearing substitutions PheBeta24Ala and PheBeta24Cys were investigated using molecular docking calculations. The Cys substitution revealed much better enantioselectivity traits with a set of seven substrates. To verify the relevance of in-silico predictions, we constructed a PGA. A + Phe24BetaCys mutant and determined its enantioselectivity in biocatalytic reactions. Since we experimentally confirmed all these predictions, we expanded our in-silico analysis to another set of seven compounds: the prediction suggested increased enantioselectivity for N-phenylacetyl-p-F-α-phenylalanine. The (R)-enantiomer of this substrate is used as a building block in synthesis of important anti-cancer agent Abarelix. The enantioselectivity of PGA. A + Phe24BetaCys mutant towards this substrate was improved by 75percent reaching E-value of about 70. Our results suggest the rapid identification of interesting replacements altering enantioselectivity using in-silico approach as the way for further expanding biotechnological application of penicillin G acylase.

Druh

J_SC - Článek v periodiku v databázi SCOPUS

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

—

Návaznosti

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2016

Kód důvěrnosti údajů

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

Název periodika

Journal of Molecular Catalysis - B Enzymatic

ISSN

1381-1177

e-ISSN

—

Svazek periodika

133

Číslo periodika v rámci svazku

Supplement 1 (November 2016)

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

7

Strana od-do

"S53"-"S59"

Kód UT WoS článku

—

EID výsledku v databázi Scopus

2-s2.0-85009773584