Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0...

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UNITN-Trento Solar-pMFC

Transcript of Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0...

Page 1: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

UNITN-Trento

Solar-pMFC

Page 2: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 3: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 4: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 5: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 6: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

PR

Page 7: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

PR

Page 8: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 9: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

araC-pBAD

araC-pBAD

Proteorhodopsin

Proteorhodopsin

Page 10: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 11: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

LIGHT DARK

Page 12: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

More H+ pumping outside!

Page 13: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

0

5

10

15

20

BBa_K731201 BBa_K1604010

Dark DarkLight Light

araC-pBAD Proteorhodopsin

ADP/ATP ratioPR increases ATP production! AD

P/AT

P ra

tio

Page 14: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 15: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 16: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

β-carotene gets used up!

β-carotene β-carotene+blh

Page 17: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

2+

Page 18: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 19: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 20: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

Abs

orba

nce

(450

nm)

1,0

2,0

3,0

4,0

LB LB LB + NATerrific Broth

Terrific Broth

pncB BBa_K1604031 araC-pBAD BBa_K731201

pncB increases NAD+ concentration!N

AD

+/N

AD

H

LB + Nicotin acid

Page 21: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

NAD

+ total

pncB produces more NAD+ without O2!

BBa_K731201 BBa_K1604010

Page 22: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 23: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 24: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 25: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 26: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 27: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

Proteorhodopsin enhances MFC performance power curve

Pow

er (μ

W)

0

3,5

7

10,5

14

Current (µA)

0,01 0,1 1 10 100 1000

BBa_K731201BBa_K1604010

Volta

ge (V

)

0

0,15

0,3

0,45

0,6

Current (µA)

0,01 0,1 1 10 100 1000

BBa_K731201 BBa_K1604010araC-pBAD Proteorhodopsin

PR enhances MFC performance

ATPATPATP

Page 28: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

pncB enhances MFC performance

Vara(V) VaraNR(V) VaraMB(V) VpncB(V) VpncBNR(V) VpncBMB(V) Iara(uA) IaraNR(uA) IaraMB(uA) IpncB(uA) IpncBNR(uA) IpncBMB(uA) Para(uA) ParaNR(uA) ParaMB(uA) IpncB(uA) PpncBNR(uA)PpncBMB(uA)

10000000 100000 0,53 0,557 0,612 0,512 0,567 0,534 0,00265 0,053 0,0557 0,0612 0,0512 0,0567 0,0534 0,000795 0,02809 585375,471698113672430,8797127470,0262144 627908,203125502920,634920635

50000 500 0,415 0,537 0,592 0,484 0,544 0,524 0,002075 8,3 10,74 11,84 9,68 10,88 10,48 0,1245 3,4445 694865,060240964652633,1471135944,68512 611438,016528926504735,294117647

25000 250 0,345 0,502 0,562 0,455 0,532 0,509 0,001725 13,8 20,08 22,48 18,2 21,28 20,36 0,207 4,761 730446,376811594629171,314741036 8,281 622030,769230769486994,360902256

20000 200 0,318 0,497 0,541 0,437 0,511 0,497 0,00159 15,9 24,85 27,05 21,85 25,55 24,85 0,2385 5,0562 776757,86163522588895,3722334 9,54845 597530,892448512483383,561643836

15000 150 0,278 0,468 0,519 0,421 0,511 0,48 0,00139 18,5333333333333 31,2 34,6 28,066666666666734,0666666666667 32 0,278 5,15226666666667787856,115107914575557,69230769211,8160666666667620239,904988124450880,626223092

10000 100 0,228 0,449 0,489 0,387 0,486 0,455 0,00114 22,8 44,9 48,9 38,7 48,6 45,5 0,342 5,1984 884214,912280702532563,47438752814,9769 610325,581395349425977,366255144

1000 10 0,056 0,185 0,258 0,127 0,244 0,223 0,00028 56 185 258 127 244 223 0,84 3,136 611160,714285714359805,40540540516,129 468787,401574803203807,37704918

500 5 0,025 0,024 0,025 0,049 0,057 0,07 0,000125 50 48 50 98 114 140 0,75 1,25 23040 26041,6666666667 4,802 66306,122448979685964,9122807018

Volta

ge (V

)

0

0,2

0,4

0,6

Current (uA)

0,01 0,1 1 10 100 1000

BBa_K731201 BBa_K1604031

Polarization curve

V

0

0,15

0,3

0,45

0,6

uA

0,01 0,1 1 10 100 1000

araNRPncbNR

Polarization curve

V

0

0,175

0,35

0,525

0,7

uA

0,01 0,1 1 10 100 1000

araMBPncbMB

Polarization curve

V

0

25000

50000

75000

100000

uA

0,01 0,1 1 10 100

araaraaraNR

Polarization curve

V

0

0,15

0,3

0,45

0,6

uA

0,01 0,1 1 10 100 1000

pncBpncBNRpncBMB

araC-pBAD pncB

NAD+NAD+NAD+

Page 29: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

QUALCHE TITOLO?

Page 30: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 31: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 32: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 33: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

NORMAL LAMP

LOW CONSUMPTIONLAMP

LED OUR SYSTEM

600 W 100 W 15 mW 200 uW

Page 34: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201
Page 35: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

In the future Synthetic Biology and household consumption must meet.

Page 36: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

Promoter strength ratio is consistent among techniques

Page 37: Solar-pMFC2015.igem.org/files/presentation/UNITN-Trento.pdf · Absorbance (450nm) 1,0 2,0 3,0 4,0 LB LB LB + NA Terrific Broth Terrific Broth pncB BBa_K1604031 araC-pBAD BBa_K731201

Attributions

Support

Wet lab: Alessandro, Claudio, Elisa, Silvia, Riccardo, Veronica. Wiki design: Moreno. Wiki content: everybody. MFC: Alessandro and Riccardo. Interlab: Claudio and Elisa. HP: Elisa. Modeling: Kent 2015 iGEM team. Logo and graphics: Gianmarco. Poster: Veronica and Elisa.

HPLC: Graziano Guella. Electronic instruments and advice: technical staff of the Physics dept. FACS and qPCR: Mansy lab. Instructors: Cristina Del Bianco, Sheref Mansy, Martin Hanczyc, Olivier Jousson.