Bunker Oil Biodesulfurization

A Feasibility Study

Presented by : Dr. Rong YAN Institute of Environmental Science and Engineering Nanyang Technological University, Singapore (email: ryan@ntu.edu.sg)

The 5th Asian Petroleum Technology Symposium Jakarta, Indonesia, January 23 to 25, 2007

Background
Singapore has a thriving maritime business, in particular bunker industry. The amount of bunker oil provided by Singapore as ship fuels is about 20 million tons annually, which is equivalent to S$ 6 billion per year. However, combustion of bunker oil has caused a significant release of gaseous pollutants (SOx), leading to marine air pollution. In Singapore, the regulation of sulfur content in bunker fuels is 4.5% on weight basis. The new regulation with a stricter limit of 1.5% sulfur might be adopted by Singapore in future, which would cause a great impact on Singapores maritime business.

Driving Force New Cap on Sulfur Contents

In 2006, the coming into force of Annex VI of the International Maritime Organization (IMO)'s MARPOL convention, as well as European Union (EU) Directive 2005/33/EC, has imposed a 1.5% sulfur cap on marine fuels used by ships in the Baltic Sea and on passenger vessels on regular services to EU community ports. In 2007, the 1.5% sulfur cap will be expanded to the second Sulfur Emissions Control Area (SECA) for the North Sea and English Channel. Both the IMO and EU legalization are, however, subject to further review in the near future, opening the possibility of further sulfur reductions. The Directive, for example, envisages a future extension of the sulfur limit to all EU waters, and also a stricter sulfur cap, possibly just 0.5%.

Why Biodesulfurization?
Currently, hydrodesulfurization (HDS) is the most widely used method in refineries to desulfurize crude oil. The HDS process requires high temperature and pressure, thus is costly and consumes lots of energy. Moreover, HDS has not been proven to remove the heterocyclic sulfur compounds like dibenzothiophene (DBT) and its derivatives efficiently. Limitations of the HDS technology can be overcome by BDS, as certain microorganisms could use organosulfur in oil as their sole energy source. Biodesulfurization (BDS) of bunker oil is expected to be a more viable option, as it is more energy efficient, and environmentally and economically favorable.

Various types of Scontaining organic compounds in crude oils

(Adapted from Shennan 1996, J. Chem. Tech. Biotechnol. )

Bunker oil (residue) mostly contains heavy molecules of organo-S with aromatic rings

However, DBT and methyl-substituted DBT (4-MDBT and 4,6-DMDBT) were particularly refractory to HDS and were not converted even at 390 C.
4th --- Prof. Atsushi Ishihara, Tokyo Univ. of Agriculture and Technology, presentation at the JPEC Asian Petroleum Technology Symposium, Jan. 2006, Cambodia

HDS and Its Limitations

Claus process: 2H2S + SO2 3S(s) + 2H2O

HDS process: It requires generally high temperature (200-425C) and pressure (150-250 psi), and consumes hydrogen extremely high energy consumption. It requires novel catalysts: (a) new support, (b) noble metal based catalyst, (c) zeolites, (d) new compositions high cost. It has not proven yet to desulfurize DBT and its derivatives no efficiency.

Alternative Option: Biocatalysts, Biodesulfurization

Objectives of Our Study

Feasibility of bunker oil biodesulfurization to biodegrade the organosulfur species present in bunker oil using suitable microorganisms (1st phase) To develop an advanced biodesulfurization process, aiming to demonstrate the biotechnology of improving bunker oil quality in the near future (2nd phase)

Scope of 1st Phase Study

I - Literature review on bunker oil, sulfur distribution in bunker oil, and the key issues related to biodesulfurization of oil II - Biodesulfurization of selected model sulfur species III - Biodegradation of organic sulfur species present in bunker oil

I. Literature Review Sulfur Biochemistry of Bunker Oil

Bunker oil contains heterocyclic sulfur compounds like thiophenes (TH), benzothiophenes (BTH), and most possibly dibenzothiophenes (DBT) and its derivatives.

Sulfur Biodegradation Pathways

Oxidative C-C cleavage Oxidative C-S cleavage (4S pathway): non destructive BDS

DBT degradation by R. erythropolis IGTS8 Source: Vazquez-Duhalt, 2002

Key Issues in Sulfur Biodegradation

(1) Selection of seed to be capable to biodesulfurize bunker oil at an improved efficiency and also with no disturbance to the organic fraction (i.e., heating value); mixed culture, isolation and bioaugmentation (2) Model sulfur species to represent bunker oil (3) Bioreactor / Culturing media (4) Temperature (bioactivity of bacteria / physical properties of water/oil system) (5) Mixing ratio of bunker oil and water / Emulsion product (6) Separation of biomass and oil (7) Others (working time, mass transfer rate, kinetic behavior etc.)

Challenges in Bunker Oil Sulfur Biodegradation

(1) Biodesulfurization of heavy oil (including bunker oil) has so far rarely been reported, although quite some researches on crude oil and diesel biodesulfurization have been conducted. (2) There is limited information on the dominant organosulfur species of bunker oils. (3) High viscosity of bunker oil could be a major concern in water/oil bio-systems. (4) Efficient separation of the three phases (biomass, oil and water) is essential but very challenging.

II. Biodesulfurization of Model Sulfur Species

As the first step of our overall experiment plan, biodesulfurization of three selected model sulfur species (TH, BTH, and DBT) were carried out, to get the suitable bacteria and familiar with essential analytical approaches.

Bacterial seed: oily sludge Culture medium: sulfur-free basal salt medium consisting of (in g L-1) KH2PO4 2.44, Na2HPO4 5.57, NH4Cl 2.00, MgCl26H2O 0.36, FeCl36H2O 0.001, MnCl24H2O 0.004 and glycerol 1.84. The final medium pH was 7.0 Instrument and identification: GCMS, IC, Plate counting, biomass and sulfur content, and TOC analyzer

Batch Reactor

Bacterial CFU number increasing

Dibenzothiophene
0.3 Concentration (mM) 0.25 0.2 0.15 0.1 0.05 0 0 2 4 Operation Tim e (d) 6 8 seed control

Change of DBT conc. in seed and control reactors

GCMS Identification of Biodesulfurization Products

Control Day 6 Control Day 0

DBT

Abundance

TIC: 021006S.D
4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000

Seed Day 0 Seed Day 6

3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.0011.0012.0013.0014.0015.00 Time-->

In seed reactor, DBT-sulfone and hydroxybiphenyl (HBP) were found with relatively high abundance at 12.466 min and 9.036 min. No DBT.

Confirmation of 4S Pathway with identifying DBT Sulfone and HBP

DBT and the intermediates
4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 0 0 2 4 operation time (d)
dibenzothiophene sulfone p-hydroxybiphenyl dibenzothiophene

GC area count

The bacteria within the mixed culture can carry out the non-destructive BDS !

Sulfur Mass Balance

Five probable sinks of sulfur: (1) non-degraded thiophenic compounds (2) organic metabolites (i.e., DBT sulfone, DBT sultine etc.) (3) soluble inorganic ions (i.e., sulfite, sulfate and thiosulfate) (4) cellular material in bacteria (5) surrounding atmosphere
Biomass Concentration
4.6

Most likely sulfur is enriched in biomass

Total sulfur content (mg) in biomass = Sulfur content per unit suspended solids (%) Suspended solids concentration in the sample (g L-1) Operation volume of the sample reactor (500 mL) 0.001 (L mL-1) 1000 (mg g-1)

Biomass Conc (g/L)

4.4 4.2 4 3.8 3.6 3.4 3.2 3 0 1 2 3 4 5 6 7 8

seed

Operation Time (day)

III. Bunker Oil Biodesulfurization

Enrichment of Bacteria & Bioreactor Setup Bunker oil to replace model sulfur species Culture medium Organosulfur solution Oil sludge Sequential batch process 1 L Erlenmeyer flask - 2 Nos (Enrichment reactor + Control) Head space aeration Bioreactor - 50 mL centrifuge tube Bunker Oil - 2 types: Bunker Oil #1 and Bunker Oil #2

Enrichment reactors cultivation of desulfurizers

Bioreactors

Experimental Approach

Schematic of experimental approach with bunker oil

Critical Parameters Adjusted

(1) Volumetric ratio of bunker oil - culture medium: 1:3 ;1:10 (2) Bunker Oil types: Bunker Oil#1 & Bunker Oil#2 (3) Biodegradation time [hrs]: 0, 48, 72 & 96

Analytical Methods
Bunker Oil: GC-MS (volatile organic sulfur); Elemental analysis (total organic combustible sulfur) Aqueous medium: ICPOES (total dissolved sulfur) Biomass: Elemental analysis (total organic combustible sulfur)

Identification of Organosulfurs in Bunker Oil

Abundance TIC: D0#1.D

Bunker Oil #1 0 hr

1800000 1600000 1400000 1200000 1000000 800000 600000 400000 200000

2.00 Time-->

4.00

6.00

8.00

10.00

12.00

14.00

16.00

18.00

Compound Methyl BTH Dimethyl BTH Trimethyl BTH Diethyl BTH DBT Methyl DBT Dimethyl DBT

Retention time [min] 7.733 8.208 8.616 8.933 9.7 10.085, 10.172 10.513~10.822

Main MS Peak 147,74,45 162,28,147 176,161,115 175,190,147,160 184,139,152 198,99,165 212,105,197

Abundance

90000 85000 80000 75000 70000 65000 60000 55000 50000 45000 40000 35000 30000 25000 20000 15000 10000 5000 0 2.00 Time-->

Ion 184.00 (183.70 to 184.30): D0#1.D

Extracted Ion Chromatogram for DBT in Bunker Oil#1 (0 hr)

4.00

6.00

8.00

10.00

12.00

14.00

16.00

18.00

Abundance Scan 2404 (9.680 min): D0#1.D 184 65000 60000 55000 50000 45000 40000 35000 30000 25000 57 71

Mass Spectrum of DBT in Bunker Oil#1 (0 hr)

m/z-->

20000 15000 10000 5000 0 20 28

43

85 139 113 127 152 165 196 218 230 246 240 260 260 282 280

97

40

60

80

100

120

140

160

180

200

220

Characterization of Sulfur in Bunker oil

Bunker oil #1 has higher S content Bunker oil #2 is heavier and has a higher viscosity

Comparison of intensities of organosulfur compounds in two Bunker Oils

Bunker Oil Biodesulfurization First Trial

Sulfur decreased 21.4%

Sulfur decreased 49.3%

 However, C/S ratio decreased, i.e., C-C cleavage happened Destructive BDS

Bunker Oil #1 (at 1:3 Volume Ratio)

Change in intensity of organosulfur species over time course for Bunker Oil #1 (Vol. ratio: Oil/Water 1:3)

Bunker Oil #1 (at 1:10 Volume Ratio)

Change in intensity of organosulfur species over time course for Bunker Oil #1 (Vol. ratio: Oil/Water 1:10) Ratio 1:3 is better than 1:10 for Bunker Oil #1

Bunker Oil #2 (at 1:3 Volume Ratio)

Change in intensity of organosulfur species over time course for Bunker Oil #2 (Vol. ratio: Oil/Water 1:3)

Bunker oil #2 (at 1:10 Volume Ratio)

Change in intensity of organosulfur species over time course for Bunker Oil #2 (Vol. ratio: Oil/Water 1:10) Ratio 1:10 is better than 1:3 for Bunker Oil #2

Elemental Analysis for Total Sulfur: Bunker Oil #1

Carbon/Sulfur Sulfur Content Ratio 1:3 is better than 1:10 for Bunker Oil #1

Elemental Analysis for Total Sulfur: Bunker Oil #2

Carbon/Sulfur Sulfur Content Ratio 1:10 is better than 1:3 for Bunker Oil #2

Elemental Analysis: Sulfur in Biomass

Key Challenge the separation of the individual phases namely bunker oil, biomass and aqueous medium (1) Cells break up during separation? (2) Oil contamination on biomass due to insufficient separation?

Sulfur content in biomass when degrading bunker Oil #2 with oil/water ratio of 1:10

Poor sulfur mass balance observed

Aqueous medium was analyzed for Total Dissolved Sulfur (TDS) using ICP-OES and no dissolved sulfur species were detected.

Conclusions
Bacteria suitable for degrading DBT and their derivatives were successfully cultured and enriched from the study using model sulfur species. Essential analytical approaches were also established and intermediates of DBT biodegradation were identified to confirm the 4S pathway. Organosulfur compounds in bunker oil like BTH, DBT and their derivatives were identified using GC-MS technique. Bunker oil biodesulfurization was confirmed highly feasible, the reduction of total sulfur content reached 49.3% in 1st trial of bunker oil biodesulfurization.

Conclusions (contd)
It is also evidenced that the biodesulfurization of bunker oil by bacteria are highly dependent on several parameters like ratio of bunker oil/culture medium, degradation time, and type of bunker oil. Bacteria desulfurized bunker oil #1 more efficiently when the volumetric oil/water ratio was maintained as 1:3 whilst bacteria desulfurized bunker oil #2 more efficiently when the ratio was maintained as 1:10, on the basis of elemental and GC-MS analysis. In bunker oil trials, the C/S ratio decreased over time for both fuel types and oil/water ratios, indicating possibly a destructive BDS. Further studies on bacterial enrichments and degradation pathways are recommended. Sulfur balance was not achieved due to most likely inefficient separation of biomass and bunker oil. Further investigation is needed to achieve effective separation.

Recommendations
A further investigation (i.e., the 2nd phase study) is highly recommended. This further study will contain:
New batch studies to further explore bunker oil BDS and to solve the problems encountered in Phase I. Investigation of BDS in a larger scale reactor operated in a semi-continuous or continuous mode. Optimization of process parameters such as temperature, mixing ratio of oil and water, separation of biomass and oil, working time, kinetic behavior, and others. A full investigation on the microbial ecology and functional diversity within the mixed culture, the isolation and characterization of bacteria, metabolic pathways, increasing the abundance of bacteria capable of non-destructive BDS in the mixed culture. A pilot scale demonstration work, targeting at a real industrial application of bunker oil biodesulfurization.

THANK YOU. And any questions?