Potensi Aktinobakteri Endofit Tabat Barito ('Ficus deltoidea Jack) sebagai Penghasil Inhibitor a-Glukosidase

Abstract

Diabetes mellitus (DM) is a chronic and serious metabolic disorder that is t revalent in the world. Diabetes mellitus occurs due to insulin deficiency and inswli resistance. This condition causes high blood glucose levels or called hYN.. rglycemia condition. One therapeutic approach to treat diabetes is to retard the abs" tion of glucose via inhibition of enzymes, such as a-glucosidase, in the small tinal. Inhibition of this enzyme slows the elevation of blood glucose following · rl:hohydrate meal. The a-glucosidase inhibitor is a bioactive compounds i8uted widely in plants and microorganisms including actino bacteria. }\ctinobacteria are Gram positive bacteria widely distributed in nature. They produce numerous metabolic products, most of which are bioactive compound with applications in many therapeutic areas. Actinobacteria are predominantly free living and are found in diverse environments including plants tissue. Microbes that live on plant tissues without causing harmfull to the host are called endophytes. Such icrobial endophytes are beginning to pay attention ofresearchers due to their abilit)1 to produce various compounds of pharmacological value similar to their li.osts. Previous reports showed that endophytic actinobacteria have a-glucosidase mhibi or activity such as Tinospora crispa (Brotowali) and Datura stramonim L (Kecubung). lf abat barito (Ficus deltoidea Jack) is a medicinal plant originating from Southeast Asia. In Indonesia, tabat barito has the most distribution in Borneo island. Tabat barito was used traditionally to treat diabetes. On another hand, several studies have shown that tabat barito has an a-glucosidase inhibitor activity. The abilit)1of tabat barito to produce a-glucosidase inhibitors is suggested to be related to the presence of endophytic microbes including actinobacteria. Previous studies showed that the abundance of actinobacteria in the tabat barito plant tissue is quite umer:ous and varied. Based on these reports, it is expected that the capability of abat barito on producing a-glucosidase inhibitors is related to the presence of actinobacteria. In this study, it was proven that endophytic actinobacteria from tabat barito <t:ould produce a-glucosidase inhibitors. Screening using a-glucosidase from rat intestine is recommended for in vitro a-glucosidase inhibitor because the homology sequeroce of catalytic side closer to enzymes in the human intestine. In this study endophytic actinobacterial isolates more inhibited a-glucosidase from rat intestines according to acarbose which only inhibit mammalian a-glucosidase. Screening of 40 enoophytic actinobacteria isolates using yeast a-glucosidase showed that 25 sG).}ates (62%) of isolates had a-glucosidase inhibitors activity. Whereas screening msm rat intestinal enzymes from 40 isolates, 31 isolates (77%) had a-glucosidase lnor activity. Aqueous extract and n-hexane ofTBL 7 isolates were the best extracts for a­ sidase inhibition, IC₅₀ value of 159.25 µg mL-¹ (aqueous extract) and 118.52 mIL-1 (n-hexane extract). Based on the 16S rRNA gene TBL 7 showed was

similar to Streptomyces tendae (99.91%). The ISP 2 medium with 10 days incubation period is the best condition to produce a-glucosidase inhibitor compound. After fractionation, AF 2 from aqueous crude extract has ICso value of 58.8 µg mL-I, higher than acarbose as positive control (ICso = 90.4 µg mL-I). The LC-MS analysis showed that the fraction of AF 2 had bioactive compounds similar t phenylpropynal, butyric acid, solketal, p-coumaric acid. The butyric acid and p­ c u aric acid have been reported to have a-glucosidase inhibitor activity. P­ c u aric acid is a pheno lie compound that has been known to have antidiabetic a..t;ivity because of its ability to reduce glucose absorption in the intestine. The p; esence of AF 2 in the reaction did not change the Km value of 1.38 mM. Whereas, t e Vmax value decreased from 0.52 mM min-I to 0.40 mM min-I. Based on these r . ults, it can be concluded that the inhibitor of the fraction is a non-competitive itili.i itor. Based on the results of this study, endophytic actinobacterial extracts can developed as antihyperglycemic agents.