Fig 2.

Purification of recombinant acrylyl-CoA reductases heterologously produced by E. coli Rosetta2 (DE3). Various fractions obtained during purification were separated on a 10% SDS-PAGE gel and stained with Coomassie blue. (A) Acrylyl-CoA reductase (AcuI) from Rhodobacter sphaeroides 2.4.1. Lane 1, whole cells before induction; lane 2, cell extract after 17 h of induction (20 μg); lane 3, recombinant AcuI eluted with E. coli GroEL (arrow) from the Ni⁺ affinity column (10 μg); lane 4, purified recombinant AcuI from the Superose 12 column (2 μg); lane 5, molecular mass standards. (B) Acrylyl-CoA reductase (product of SPO_1914) from Ruegeria pomeroyi DSS-3. Lane 1, whole cells before induction; lane 2, cell extract after 17 h of induction (20 μg); lane 3, recombinant product of SPO_1914 from the Ni⁺ affinity column (4 μg); lane 4, purified recombinant product of SPO_1914 from the Superose 12 column (4 μg); lane 5, molecular mass standards. (C) Acrylyl-CoA reductase (YhdH) from Escherichia coli K-12 substrain MG1655. Lane 1, molecular mass standards; lane 2, whole cells before induction; lane 3, cell extract after 18 h of induction (20 μg); lane 4, recombinant YhdH from the Ni⁺ affinity columns (4 μg); lane 5, purified recombinant YhdH from the Superose 12 column (2 μg).