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Synthesis of potential pharmaceutical active ingredients using omega-transaminase

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2017
03_biochem2017.pdf (529.8Kb)
Autori
Marković, Nevena
Jovanović Šanta, Suzana
Prodanović, Radivoje
Konferencijski prilog (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentu
Apstrakt
Transaminases (EC 2.6.1.X) are enzymes which catalyze reversible transfer of amino group from amino acids to α-keto acids by using piridoxal-5ʼ-phosphate as a coenzyme. There is a huge interest for the application of ω-transaminases in industrial production of chiral amines and alkaloids since those compounds are extensively used in pharmaceutical, agricultural, and chemical industries. Application of ω-transaminases in asimetric synthesis of these compounds enables efficient production of biologicaly active amines, due to their catalytic properties for synthesis with a high level of enantioselectivity, supstrate promiscuity (they are capable to aminate keto acids, aldehydes and ketones), high turnover number, no requirement for regeneration of external cofactors, and among other cheaper, simpler and green process of production. We are developing biocatalytic route for the synthesis of amino steroids by using ω- transaminase, (R)-selective, ATA-117 enzyme variant from Arthrobacter sp 3.... It can be observed that enzyme expression was done in Echerichia coli BL21 D3 pLysS (Figure 1), and HPLC analysis of enzyme activity and specificity toward 15 structuraly different steroid compounds was performed. (R)-methylbenzylamine was used as amino group donor and pyridoxal-5ʼ-phosphate as cofactor. Activity of the enzyme was measured in bacterial lysate based on the absorbance of acetophenone, that is formed during the transamination reaction of (R)-methylbenzylamine. Figures 2 and 3 are showing chromatograms of acetophenone standard and products of reaction performed with enzyme expressed in E. coli and 16,17-epoxypregnenolone. Reactions were analysed on reversed phase column NucleosilC18. Based on the results, we have selected four steroid compounds for which enzyme showed highest activity and with a potential for biological activity. The next step was optimisation of the reaction conditions with a low cost amino donor isopropylamine, and isolation and characterisation of a pure amino steroid products. Until now we have managed to enzymatically synthesize and purify one amino steroid which should be further analysed by spectral characterization and its biological activity will be determined.

Ključne reči:
Transaminases / biocatalytic route / synthesis of amino steroids
Izvor:
Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings, 2017
Izdavač:
  • Faculty of Chemistry, Serbian Biochemical Society
Projekti:
  • Ispitivanja odnosa struktura-funkcija u ćelijskom zidu biljaka i izmene strukture zida enzimskim inženjeringom (RS-173017)
[ Google Scholar ]
URI
http://www.bds.org.rs/en/conferences.php
http://cer.ihtm.bg.ac.rs/handle/123456789/3281
Kolekcije
  • Radovi istraživača / Researchers' publications
Institucija
IHTM
TY  - CONF
AU  - Marković, Nevena
AU  - Jovanović Šanta, Suzana
AU  - Prodanović, Radivoje
PY  - 2017
UR  - http://www.bds.org.rs/en/conferences.php
UR  - http://cer.ihtm.bg.ac.rs/handle/123456789/3281
AB  - Transaminases (EC 2.6.1.X) are enzymes which catalyze reversible transfer of amino group from amino acids to α-keto acids by using piridoxal-5ʼ-phosphate as a coenzyme. There is a huge interest for the application of ω-transaminases in industrial production of chiral amines and alkaloids since those compounds are extensively used in pharmaceutical, agricultural, and chemical industries. Application of ω-transaminases in asimetric synthesis of these compounds enables efficient production of biologicaly active amines, due to their catalytic properties for synthesis with a high level of enantioselectivity, supstrate promiscuity (they are capable to aminate keto acids, aldehydes and ketones), high turnover number, no requirement for regeneration of external cofactors, and among other cheaper, simpler and green process of production. We are developing biocatalytic route for the synthesis of amino steroids by using ω- transaminase, (R)-selective, ATA-117 enzyme variant from Arthrobacter sp 3. It can be observed that enzyme expression was done in Echerichia coli BL21 D3 pLysS (Figure 1), and HPLC analysis of enzyme activity and specificity toward 15 structuraly different steroid compounds was performed. (R)-methylbenzylamine was used as amino group donor and pyridoxal-5ʼ-phosphate as cofactor. Activity of the enzyme was measured in bacterial lysate based on the absorbance of acetophenone, that is formed during the transamination reaction of (R)-methylbenzylamine. Figures 2 and 3 are showing chromatograms of acetophenone standard and products of reaction performed with enzyme expressed in E. coli and 16,17-epoxypregnenolone. Reactions were analysed on reversed phase column NucleosilC18. Based on the results, we have selected four steroid compounds for which enzyme showed highest activity and with a potential for biological activity. The next step was optimisation of the reaction conditions with a low cost amino donor isopropylamine, and isolation and characterisation of a pure amino steroid products. Until now we have managed to enzymatically synthesize and purify one amino steroid which should be further analysed by spectral characterization and its biological activity will be determined.
PB  - Faculty of Chemistry, Serbian Biochemical Society
C3  - Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings
T1  - Synthesis of potential pharmaceutical active ingredients using omega-transaminase
ER  - 
@conference{
author = "Marković, Nevena and Jovanović Šanta, Suzana and Prodanović, Radivoje",
year = "2017",
url = "http://www.bds.org.rs/en/conferences.php, http://cer.ihtm.bg.ac.rs/handle/123456789/3281",
abstract = "Transaminases (EC 2.6.1.X) are enzymes which catalyze reversible transfer of amino group from amino acids to α-keto acids by using piridoxal-5ʼ-phosphate as a coenzyme. There is a huge interest for the application of ω-transaminases in industrial production of chiral amines and alkaloids since those compounds are extensively used in pharmaceutical, agricultural, and chemical industries. Application of ω-transaminases in asimetric synthesis of these compounds enables efficient production of biologicaly active amines, due to their catalytic properties for synthesis with a high level of enantioselectivity, supstrate promiscuity (they are capable to aminate keto acids, aldehydes and ketones), high turnover number, no requirement for regeneration of external cofactors, and among other cheaper, simpler and green process of production. We are developing biocatalytic route for the synthesis of amino steroids by using ω- transaminase, (R)-selective, ATA-117 enzyme variant from Arthrobacter sp 3. It can be observed that enzyme expression was done in Echerichia coli BL21 D3 pLysS (Figure 1), and HPLC analysis of enzyme activity and specificity toward 15 structuraly different steroid compounds was performed. (R)-methylbenzylamine was used as amino group donor and pyridoxal-5ʼ-phosphate as cofactor. Activity of the enzyme was measured in bacterial lysate based on the absorbance of acetophenone, that is formed during the transamination reaction of (R)-methylbenzylamine. Figures 2 and 3 are showing chromatograms of acetophenone standard and products of reaction performed with enzyme expressed in E. coli and 16,17-epoxypregnenolone. Reactions were analysed on reversed phase column NucleosilC18. Based on the results, we have selected four steroid compounds for which enzyme showed highest activity and with a potential for biological activity. The next step was optimisation of the reaction conditions with a low cost amino donor isopropylamine, and isolation and characterisation of a pure amino steroid products. Until now we have managed to enzymatically synthesize and purify one amino steroid which should be further analysed by spectral characterization and its biological activity will be determined.",
publisher = "Faculty of Chemistry, Serbian Biochemical Society",
journal = "Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings",
title = "Synthesis of potential pharmaceutical active ingredients using omega-transaminase"
}
Marković N, Jovanović Šanta S, Prodanović R. Synthesis of potential pharmaceutical active ingredients using omega-transaminase. Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings. 2017;
Marković, N., Jovanović Šanta, S.,& Prodanović, R. (2017). Synthesis of potential pharmaceutical active ingredients using omega-transaminase.
Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - ProceedingsFaculty of Chemistry, Serbian Biochemical Society..
Marković Nevena, Jovanović Šanta Suzana, Prodanović Radivoje, "Synthesis of potential pharmaceutical active ingredients using omega-transaminase" (2017)

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