ENHANCEMENT OF BIODESULFURIZATION IN RHODOCOCCUS SPECIES (IGTS8) BY THE EXPRESSION OF VITREOSCILLA HEMOGLOBIN

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Description

The bacterium Rhodococcus sp. IGTS8 contains the dsz operon, which encodes a three enzyme pathway (the “4S pathway”) that is able to mineralize the sulfur contained in dibenzothiophene (DBT), an organic sulfur containing molecule found in petroleum. The gene vgb, which encodes Vitreoscilla... Show moreThe bacterium Rhodococcus sp. IGTS8 contains the dsz operon, which encodes a three enzyme pathway (the “4S pathway”) that is able to mineralize the sulfur contained in dibenzothiophene (DBT), an organic sulfur containing molecule found in petroleum. The gene vgb, which encodes Vitreoscilla hemoglobin (VHb), has shown wide usefulness in enhancing productivity and other useful properties when expressed in heterologous hosts. We engineered strain IGTS8 to express VHb and measured the effects on growth and desulfurization of DBT, using minimal medium containing DBT as the sole source of sulfur. VHb was clearly detected in the engineered strain using the standard COdifference spectral analysis, but its level (0.38-0.63 nmoles/gm wet weight of cells) was about 10-fold lower than commonly seen for expression of VHb in other heterologous bacterial hosts. The VHb-expressing strain was tested for growth at both low and high aeration in minimal medium containing DBT as sole sulfur source; growth was about 50% lower at low aeration compared with high aeration. Despite this, metabolism of DBT (as detected by accumulation of the end product of the 4S pathway, 2-Hydroxy biphenyl (2-HBP), in the growth medium) was about 30 % higher in the low aeration compared to the high aeration culture. A possible explanation for these results is direct enhancement of the first two (monooxygenase) steps in conversion of DBT to 2-HBP. It was thus concluded from the studies that the expression of vgb in Rhodococcus sp. IGTS8 enhances the process of biodesulfurization under conditions of low aeration
M.S. in Biology, July 2013 Show less

Details
Department

BIOL / Biology

Title

ENHANCEMENT OF BIODESULFURIZATION IN RHODOCOCCUS SPECIES (IGTS8) BY THE EXPRESSION OF VITREOSCILLA HEMOGLOBIN

Institution

Illinois Institute of Technology

Creator(s)

Shivdas, Vrushali D.

Advisor(s)

Stark, Benjamin

Date

2013
2013-07

Description

The bacterium Rhodococcus sp. IGTS8 contains the dsz operon, which encodes a three enzyme pathway (the “4S pathway”) that is able to mineralize the sulfur contained in dibenzothiophene (DBT), an... Show moreThe bacterium Rhodococcus sp. IGTS8 contains the dsz operon, which encodes a three enzyme pathway (the “4S pathway”) that is able to mineralize the sulfur contained in dibenzothiophene (DBT), an organic sulfur containing molecule found in petroleum. The gene vgb, which encodes Vitreoscilla hemoglobin (VHb), has shown wide usefulness in enhancing productivity and other useful properties when expressed in heterologous hosts. We engineered strain IGTS8 to express VHb and measured the effects on growth and desulfurization of DBT, using minimal medium containing DBT as the sole source of sulfur. VHb was clearly detected in the engineered strain using the standard COdifference spectral analysis, but its level (0.38-0.63 nmoles/gm wet weight of cells) was about 10-fold lower than commonly seen for expression of VHb in other heterologous bacterial hosts. The VHb-expressing strain was tested for growth at both low and high aeration in minimal medium containing DBT as sole sulfur source; growth was about 50% lower at low aeration compared with high aeration. Despite this, metabolism of DBT (as detected by accumulation of the end product of the 4S pathway, 2-Hydroxy biphenyl (2-HBP), in the growth medium) was about 30 % higher in the low aeration compared to the high aeration culture. A possible explanation for these results is direct enhancement of the first two (monooxygenase) steps in conversion of DBT to 2-HBP. It was thus concluded from the studies that the expression of vgb in Rhodococcus sp. IGTS8 enhances the process of biodesulfurization under conditions of low aeration Show less

Type

Thesis

File type

application/pdf

Access

Restricted Access

Rights

In Copyright
http://rightsstatements.org/page/InC/1.0/

Identifier

http://hdl.handle.net/10560/3157

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