Abstract
Betulinic acid is a product of plant secondary metabolism which has shown various bioactivities. Several CYP716A subfamily genes were recently characterized encoding multifunctional oxidases capable of C-28 oxidation. CYP716A12 was identified as betulin C-28 oxidase, capable of modifying betulin. This study aimed to induce the transformation of betulin to betulinic acid by co-expressing enzymes CYP716A12 from Medicago truncatula and ATR1 from Arabidopsis thaliana in Saccharomyces cerevisiae. The microsome protein extracted from the transgenic yeast successfully catalyzed the transformation of betulin to betulinic acid. We also characterized the optimization of cell fragmentation, protein extraction method, and the conversion conditions. Response surface methodology was implemented, and the optimal yield of betulinic acid reached 18.70%. After optimization, the yield and the conversion rate of betulin were increased by 83.97% and 136.39%, respectively. These results may present insights and strategies for the sustainable production of betulinic acid in multifarious transgenic microbes.
Highlights
Triterpenoids are a diverse class of chemical compounds, and they have been increasingly attracting the attention of researchers due to their beneficial bioactivities in the health field.Betulinic acid (3-hydroxy-lup-20(29)-en-28-oic acid) is a pentacyclic lupane-type triterpenoid that is widely distributed throughout the entire plant kingdom
Among these compounds, betulinic acid has gained considerable interest owing to a variety of biological and pharmacological activities that have been ascribed to this compound including anti-inflammatory, antibacterial, antiviral, antimalarial, anti-HIV, and antitumor effects [1]
An alternative approach for producing betulinic acid from betulin was achieved by a biotransformation process, but the conversion efficiency was rather poor, and this metabolic pathway is subject to the restriction of limited betulin supply [4]
Summary
Triterpenoids are a diverse class of chemical compounds, and they have been increasingly attracting the attention of researchers due to their beneficial bioactivities in the health field.Betulinic acid (3-hydroxy-lup-20(29)-en-28-oic acid) is a pentacyclic lupane-type triterpenoid that is widely distributed throughout the entire plant kingdom. Among these compounds, betulinic acid has gained considerable interest owing to a variety of biological and pharmacological activities that have been ascribed to this compound including anti-inflammatory, antibacterial, antiviral, antimalarial, anti-HIV, and antitumor effects [1]. Despite its extraordinary potential for therapeutic applications, the insufficient sources of betulinic acid in plants is a major challenge in commercializing this therapeutic compound. An alternative approach for producing betulinic acid from betulin was achieved by a biotransformation process, but the conversion efficiency was rather poor, and this metabolic pathway is subject to the restriction of limited betulin supply [4]. Rapid progress in synthetic biology and metabolic engineering provides another way to achieve a high yield of natural
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