The prevalence of obesity and diabetes in the United States and globally has been rising in the last few decades and it continues to climb. One of the causes for this increase is the over consumption of sugars and/or carbohydrates. Among medications available to treat obesity and diabetes are the α-glucosidase (α-amylase) inhibitors. This class of medication decreases carbohydrate hydrolysis in the intestines, leading to reduced glucose absorption. Trestatins are a group of α-amylase inhibitors produced by the soil bacterium Streptomyces dimorphogenes. Despite their high pharmaceutical potential, little is known about their modes of formation in nature. Therefore, the main objective of this study is to identify the genes involved in trestatin biosynthesis and to understand their roles in the pathway. To confirm the production of trestatin in S. dimorphogenes, the bacterium was grown on R2A agar plates and cultured in a liquid production medium for 4.5 days. The culture broth was passed through an ion-exchange [Dowex 50Wx2 (H+ form)] column and the lyophilized crude product was analyzed by mass spectrometry. To create mutant strains of S. dimorphogenes, two plasmids, pTMAD003 and pTMAD004, were constructed using Gibson Assembly and subsequently transferred into S. dimorphogenes through conjugation. However, the conjugation experiments have so far been unsuccessful. Alternative methods for gene transfer and inactivation are currently being investigated.