Difference between revisions of "Part:BBa K4765118"

Revision as of 06:38, 12 October 2023

ribozyme connected: MysABCDH

Introduction

Mycosporine-like amino acids (MAAs), act as a sunscreen for the biofilm. This composite part contains five enyzmes in the MAA biosynthesis pathway. All the enzymes are constructed into ribozyme-assisted polycistronic co-expression system：pRAP.

Figure 1. The biosynthetic pathway of shinorine, porphyra-334, palythine-Ser, and palythine-Thr

Usage and Biology

Biosynthetic route of MAA initiates with the generation of 4-deoxygadusol (4-DG) from sedoheptulose 7-phosphate, an intermediate within the pentose phosphate pathway. This process is catalyzed by two enzymes: a dimethyl 4-degadusol synthase (DDGS; MysA) and an Omethyltrans-ferase (O-MT; MysB). Subsequently, 4-DG undergoes a transformation into mycosporine-glycine(MG) through an ATP-grasp enzyme MysC, which introduces an amino acid moiety, primarily L-Gly. MAA analogues such as shinorine or porphyra-334 are further derived from MG by the D-Ala-D-Ala ligase-like enzyme MysD. In the final step, the biosynthesis is completed with a nonheme iron-(II)- and 2oxoglutarate-dependent (Fe/2OG) oxygenase MysH, leading to the production of palythines.

Characterization

Agarose gel electrophoresis

Figure 1. Agarose gel electrophoresis of PCR products, amplified from bacterial colonies/cultures. From right lane(4) to left lane(1) indicate the successful construction of MysD, MysDH, MysDHB, and MysDHBA.

Sequence and Features

Reference