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Characterization of Desulfinase for the Removal of Recalcitrant Sulfur Compounds in Petroleum
Author(s) -
Dodd Cameron
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.02632
Subject(s) - dibenzothiophene , organosulfur compounds , chemistry , sulfur , chromatography , enzyme , flue gas desulfurization , affinity chromatography , size exclusion chromatography , substrate (aquarium) , organic chemistry , biology , ecology
Biodesulfurization provides a method to remove the sulfur from organosulfur compounds found in petroleum. This multi‐step pathway is able to remove sulfur from dibenzothiophene (DBT), one of the most recalcitrant compounds found in petroleum and crude oil. A critical enzyme in this pathway is an aromatic desulfinase (DszB) that catalyzes the final rate‐limiting step of the desulfurization. The DszB enzyme from Nocardia asteroides A3H1 was overexpressed in E. coli, purified using Ni 2+ ‐affinity chromatography and characterized kinetically. Kinetic assays revealed a sigmoidal response when the velocity was plotted against substrate concentration, calling into question the monomeric structure of the enzyme previously reported by crystallography studies. Size exclusion chromatography and cross‐linking experiments were conducted to further investigate the oligomerization of HPBS desulfinase. Attempts to further purify the enzyme using Co 2+ ‐affinity chromatography provided further evidence that under certain conditions DszB may exhibit cooperativity. Support or Funding Information JMU Department of Chemistry and Biochemistry Dr. Linette M. Watkins NSF Grant No. CHE‐1757874