2/24/20

1

Life Make Rock:4 Billion Years ofBiomineralization

Bruce W. FoukeDirector Roy J. Carver Biotechnology Center

Professor of Geology and MicrobiologyCarl R. Woese Institute for Genomic Biology

Sivaguru et al. (2019) Nature Reviews Urology

2/24/20

2

Oil Field GeobiologyCO2 Sequestration

BiosouringShale Fracking

Decatur, IL USACO2 Sequestration Well

2/24/20

3

Schlumberger Quicksilver Probe

2/24/20

4

2/24/20

5

50 µm

5 µm

2/24/20

6

Actyl-CoA Benzoate degradation

Benzoate

Benzoyl-CoA Cyclohexane-1-

carboxylate Crotonoyl-CoA

(S)-3-Hydroxy-butanoyl-CoA

Acetoacetyl -CoA Acetate Flagellum

Protein, amino Acids

Fatty acids Sugars

Ribonu- cleotide

Sidera-phore

Ferric iron- coated sandstone

Ox. Red

Iron Reduction

ATP

ADP+Pi

ATP synthesis

F0F1-ATP synthase

Formation water

Serpentin- ization

Formate H2+CO2

HCO3-

HCO3-

Aromatic compounds

Metals, toxin, natural

antibiotics

Metals (e.g., Mn, Zn, Ni, Mo, Se, Wo)

Fe(III)

Fe(II) Biosyn- thesis

H+

Type IV pili

H+ Na+/K+/Ca2+

Nitrate

NO2- Nitrate

Reduction

NH4+

Glutamine Glutamate

NH4+

Ammonia Assimilation

Input signal

choline, betaine

Ectoine

5-OH -ectoine

Nγ- NAcDABA

DABA

Nα- NAcDABA

ASA

Asp

Choline!

Betaine aldehyde!

Glycine betaine!Osmosis

Response

Proteins/polysaccharide

K+!ATP

ADP

Restriction modification

foreign DNAs

M!Host DNA

M!

Biofilm

ATP!

P!PhoB

?!P! PhoR

Urea

Purine Degradation

Allanoin

Allantoate

Ureidoglycolate

D-Glycerate Pathway

Purine Utilization

H+

Output signal?

GPD

G3P G3P

G6P

F6P

PGALD

PEP

G3P

pyruvate

TCA/rTCA

6-P-gluconate

Phospholipids degradation

Aromatic compounds degradation

Succiny l-CoA

Lipids degradation

PPP ED

Acetyl -CoA

Glycolysis

R!

Halomonas sulfidaeris-dominated microbial communities significantly enriched in several metabolic categories, including: 1) virulence; 2) metal oxidation; 3) metabolism of aromatic compounds

2/24/20

7

0

5

10

15

20

25

2000 2010 2020 2030 2040 2050

AEO2020 crude oil production(million barrels per day)

tight oil

AlaskaGulf of Mexicoother

2019history projections

0

10

20

30

40

50

60

2000 2010 2020 2030 2040 2050

AEO2020 dry natural gas production by typetrillion cubic feet

Reference case

tight/shale gasotherLower 48 onshoreLower 48 offshoreother

2019history projections

~59% from tight oil

Tight oil reaches 73% of U.S. crude oil

production by 2050

Tight/shale gas reaches 91% of U.S. domestic gas natural

gas production in 2050~62% from

tight/shale gas

Data from Annual Energy Outlook 2020 infographics

Unconventional Tight/Shale Oil Gas ProductionDriver of U.S. Crude Oil and Natural Gas Production

2/24/20

8

Current Oilfield Assets

10-9

10-8

10-7

10-6

10-5

10-4

10-3

10-2

10-1

100

101

102

103

104

105Observations and Sampling

1 m

1 cm

1 mm

1 µm

1 nm

1 km

Laboratory Analyses

field

wor

k

oil f

ield

s

mic

robi

al c

ell

trans

mitt

ed li

ght

x-ra

y co

mpu

ter t

omog

raph

y

x-ra

y sc

atte

ring

Hum

an E

ye D

etec

tion

Inst

rum

ent D

etec

tion

10-3

10-2

OVERLAP

Microbially Enhanced Hydrocarbon Recovery: Relevant Length Scales

3D se

ism

ic

core

mat

eria

lwire

line

logs

flow

-thro

ugh

colu

mns

core

plu

gs10 cm

10 m

100 m

10 km

100 km

Geo

BioC

ell-R

ail S

cope

mod

elin

g

2/24/20

9

GeoBioCellMicrofluidic Testbed

GeoBioCell Experimental ApproachMicrofluidic cells to simulate

subsurface hydrodynamic environments

The GeoBioCell:Si-etched homogeneous flow

cell

Reservoir Rock-GeoBioCell:Reservoir rock core thin

section flow cells

Outlets or Inlets

Outlet or Inlet

Real rock thin section

2/24/20

10

Real Reservoir Rock-GeoBioCell

19

Reservoir Rock

GeoBioCell

PDMS (~3 mm)

Sandstone Core (0.5 mm)PDMS (~0.5 mm)

Glass (0.5 mm)

Plexy Glass (0.5 mm)

Cross-section view

Aluminum Manifold Sandstone rock core

Inlet/Outlet Ports

2.5 ns (Crude Oil)2.3 ns (Light oil)

Fluorescent Lifetime Imaging Microscopy: Crude and Light Oil Characterization in GBC

• Crude oil and light oil can be separated based on different life times, 2.5 ns and 2.3 ns, respectively.

• Labeled bacteria (e.g. Propidium iodide, lifetime 10 – 20 ns) can be imaged in crude oil in GBC

2/24/20

11

Thin Sections Impregnated with Oil and Pore Structure Analyzed with

Confocal Microscopy• Crude oil impregnated

Bentheimer core• Imaging depth ~ 400 μm• Well pore connectivity Plan and side views

3D confocal reconstruction

Particle Image Velocimetry (PIV)

2/24/20

12

8,467 ft

Mt. Simon Sandstone Reservoir Rock Analyses

24

2/24/20

13

Training at the Blind Pig

2/24/20

14

Mechanisms for Biosouring

• Biological reduction of SO42- to Sulfides (S2-, HS-, and

H2S) via sulfate reducing Bacteria (SRB)

SRB

SO42- S2-, HS-, H2S

Oil Organics CH3COO- + CO2

Corrosion

Fe > Fe2+ + 2e-

H2S + 2e- > S2- + H2

SRB

SO42-

H2S

Oil Organics

CH3COO-

+ CO2

PrRB

N2

NO3-

S0

NO3 Injection• Prevent biological reduction of SO4

2- to Sulfides (S2-, HS-, and H2S) by feeding NO3 to subsurface bacteria and thus preventing sulfate reducing Bacteria (SRB)

Cretaceous Western Interior Seaway

Well F

B’GR

FeetTVDSS

Resistivity SWCoreGR

FeetTVDSS

Resistivity SWCore

M70 and Lower M80 Reservoirs

Upper M80 Reservoirs

2/24/20

15

Upper Devonian Grosmont Limestone ReservoirUnconventional Bitumen and Heavy Oil Reservoir

>318 Billion BBL (perhaps 400-500 Billion BBL total)

Core to Pore

2/24/20

16

2/24/20

17

2/24/20

18

Deep Subsurface Microbial Biosphere, Heavy Oil, North Slope Alaska

2/24/20

19

Marathon Oil CompanyAnimation of Hydraulic Fracturing (Fracking)

https://www.youtube.com/watch?v=VY34PQUiwOQ&feature=youtu.be

2/24/20

20

Well

Oil Well

Fractures

2-5Km depth

Importance of Real Rock GeoBioCell Microfluidic Device

Shal

e

Inlet 1 Inlet 2

Outlet 2Outlet 1

Proppants

Proppants

Shale Formation

Shale Formation

Sulfate Reducing Bacteria (SRB) CulturingSRB Enrichment• Enriched from produced formation waters at 1-1.5km

depth. • Cultured in media formulated to simulate 45°C

subsurface conditions.Selective media containing volatile fatty acids (VFA) and vitamins

1. Acetate (13mM)2. Formate (15mM)3. Butyrate (1.2mM)4. Propionate (1.4mM)

Control SRB enrichment

2/24/20

21

Ecology, Physiology and Metabolism of theSubsurface Biosphere Inhabiting Fracked Shale

Real-Rock Shale GeoBioCell Construction and OperationShale GBC Shale GBC with proppants

Outlet 1

Outlet 2

Digital Pressure Gauge

GBC

Drain

Culture

Media

Drain

GBC

2/24/20

22

2/24/20

23

2/24/20

24

2/24/20

25

2/24/20

26

Shale

Flow Direction

59-71 hours of Shale GeoBioCell operation

Shale

Time Lapse Videos of Sulfate Reducing Bacteriawithin Shale Microfractures in a Microfluidic GeoBioCell

2/24/20

27

Flow Direction

Filaments

Proppants

Shale

Shale

Flow DirectionDay 6

Filaments

Proppants

Shale

Shale

2/24/20

28

Oil Field GeobiologyCO2 Sequestration

BiosouringShale Fracking