The global abundance and availability of oat hulls make them a promising feedstock to produce a unique type of cellulose, the bacterial one. This is the first study examining how a chemical pretreatment method of oat hulls influences the yield and properties of bacterial cellulose (BC) in extended cultivation. Here we employed our own pretreatment methods that use dilute HNO3 and NaOH solutions in one and two stages, a total of four pretreatment methods. Further technological stages were performed in the same manner: pulps were enzymatically hydrolyzed with commercial enzymes CelloLux-A and BrewZyme BGX, and biosynthesis of BC was run using the Medusomyces gisevii Sa-12 symbiotic culture. A two-stage (HNO3 + NaOH) pretreatment of oat hulls was found to afford a biologically good medium and increase the BC yield 1.8−3.2-fold compared to the other pretreatments used. A pretreatment method of oat hulls determined the BC yield and degree of polymerization. However, a pretreatment method had no impact on the highest crystallinity index and allomorph Iα content of all the BC samples, which is explained by Medusomyces gisevii Sa-12 used. The crystallinity index and allomorph Iα content, as measured by X-ray diffractometry, are proposed for use as BC quality assessment criteria.