Surface display of bacterial tyrosinase on spores of Bacillus subtilis using CotE as an anchor protein

Abstract

Tyrosinases, copper-containing monooxygenases, are widely used enzymes for industrial, medical, and environmental applications. We report the first functional surface display of Bacillus megaterium tyrosinase on Bacillus subtilis spores using CotE as an anchor protein. Flow Cytometry was used to verify surface expression of tyrosinase on the purified spores. Moreover, tyrosinase activity of the displayed enzyme on B. subtilis spores was monitored in the presence of L-tyrosine (substrate) and CuSO (inducer). The stability of the spore-displayed tyrosinase was then evaluated after 15 days maintenance of the spores at room temperature, and no significant decrease in the enzyme activity was observed. In addition, the tyrosinase-expressing spores could be repeatedly used with 62% retained enzymatic activity after six times washing with Tris-HCl buffer. This genetically immobilized tyrosinase on the spores would make a new advance in industrial, medical, and environmental applications.

Progress in research and application development of surface display technology using Bacillus subtilis spores

“…Tyrosinase activity on spores was monitored in the presence of L-tyrosine and CuSO 4 . Recombinant spores could be used repeatedly, with 62% of enzymatic activity remaining after washing six times with Tris-HCl buffer (Hosseini-Abari et al 2016).…”

Surface display system for probiotics and its application in aquaculture

“…In addition, cotE (Hosseini‐Abari et al . 2016), cotX (Wang et al . 2016a), cotY and cotZ (Bartels et al ., 2018) are also used as the surface anchoring protein to mediate spore surface display.…”

Metabolic Engineering of Microorganisms for the Production of Flavonoids

“…Interestingly, when using daidzein as the substrate, three orthohydroxylated daidzein derivatives were obtained, including 8-hydroxydaidzein (8-OHDe), 3 -hydroxydaidzein (3 -OHDe), and 6-hydroxydaidzein (6-OHDe), with a conversion rate of 2.4, 0.9, and 36.3%, respectively (Chang et al, 2013). In order to improve the catalytic efficiency of hydroxylase and avoid by-products formation, a tyrosinase from Bacillus megaterium was functionally displayed on Bacillus subtilis spores using CotE as an anchor protein (Hosseini- Abari et al, 2016). This spore displayed tyrosinase is able to efficiently hydroxylate 1 mM genistein into 1 mM orobol with a 100% conversion rate in 90 min (Abari and Tayebi, 2019).…”