Bioelectrocatalysis

Arkady A. KaryakinArkady A. Karyakin

Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, RussiaRussia

Bioelectrocatalysisis an acceleration of electrode reactions by biological catalysts

Enzymes Whole cells

Applications of bioelectrocatalysis

Fuel electrodes(biofuel cells)

Biosensors

Electrosysthesis

Enzyme bioelectrocatalysis

BIOELECTROCATALYSIS

S2P2

How to involve enzymes in bioelectrocatalysis?

Use of mediators:

Direct bioelectrocatalysis:

e

S

P-

S u b stra te O x id izedS u b stra te

O xid o red u cta se

M M redox

E le c tro d e

Direct bioelectrocatalysisif electrochemistry is determined by the catalyzed reaction or/and redox activity of biocatalyst

A nalyte O xid izedA nalyte

O H O2 2 2

O xidase

Oxidase catalysis

N

N

N

N O

O

H H

O

O

N

N

N

N

H

H

2e-

2H+

O

HO OH

N

N N

N

NH2

N

HN

N

N

O

O

CH3

CH3

CH2

C

C

C

H2C

HO H

HHO

HHO

O P

O

ONa

O P

O

ONa

O CH2

Redox activity of oxidases

E ≈ -0.064 В (NHE)

FAD

Mediated bioelectrocatalysis – II generation biosensors

G lu co se G lu co n icac id

G lu co se O xid a se

F c F c

E lec tro d e

+

A. E. G. Cass, G. Davis, G. D. Francis, H. A. O. Hill, W. G. Aston, I. J. Higgins, E. V. Plotkin, L. D. L. Scott, and A. P. F. Turner, Analytical Chemistry 56, 667-671 (1984).

Fe

Glucose tests• Accu-Chek Complete BG System(Boehringer Mannheim)

• Accu-Chek Easy(Boehringer Mannheim)

• Accu-Chek Instant(Boehringer Mannheim)

• Accu-Chek Instant Plus(Boehringer Mannheim)

• Autolet® II Clinisafe(Owen Mumford)

• Autolet® Lite Starter Pack(Owen Mumford)

• Blood Glucose Strips(Roche)

• Exatech®(Medisense)

• Fingerstix Lancets(Bayer)

• Glucofilm™ Test Strips(Bayer)

• Glucose Control Solution(Roche)

• Glutose®(Roche)

• Lifescan One Touch® Basic™ System(Johnson & Johnson)

• Medipoint Blood Lancets(Medipoint)

• Monolet Lancet(Kendall-Sherwood)

• Soft-Touch® II(Boehringer Mannheim)

• Softclix(Roche)

• Unilet Long-Body™ Lancets(Owen Mumford)

• Unistik™-2(Owen Mumford)

B.A. Gregg, A. Heller. Anal. Chem. 62 (1990) 258

Mediated bioelectrocatalysis – II generation biosensors

G lu co se

G lu c . a c .

O s+ /2+

O s+ /2+

O s+ /2+

hyd ro g e le_O s

+ /2+

O s+ /2+

Wiring of glucose oxidase

Heller, A. Physical Chemistry Chemical Physics 2004, 6, 209-216.

E = -0.195 mV (Ag|AgCl)

Glucose test

Therasense:0.3 µL of blood

Dehydrogenase catalysis

S u b s tra te P ro d u c tD ehydrogenase

N A D (P )HN A D (P )+

> 500 enzymes

N

R

C

O

NH2

N

R

C

O

NH2

2e-

H++

NAD+|NADH redox reaction

the lowest potential in aerobic organisms; on bare electrodes the overvoltages exceed 1 Volt.

N

SNH3C

H3C

N CH3

CH3

N CH3

CH3NO2

N

SNH3C

H3C

N CH3

CH3

N

SH2N

Methylene Blue

Methylene Green

Azur A

N CH3

CH3

N

SH2N

H2C Toluidine Blue

N CH2

CH2

CH3

CH3

N

OH2N

H2C Brilliant Cresyl Blue

N CH3

CH3

N

NH2N

H2C

H

Neurtal Red

NH2

N

SH2N

Thionine

Electropolymerized azines: a new class of electroactive polymersMethylene Blue

E, V

-0.4

0.6 1.2

0.5 mA/cm2

Neutral Red

-0.8

0.8E, V

0.1 mA/cm2

Toluidine Blue

E, V

0.1mA/cm

-0.4

0.4 0.8

2

N CH3

CH3

N

SNH3C

H3C

N CH3

CH3

N

SNH3C

N CH3

CH3

N

SNH3C

H

Hypothesis on polyazine structure

A.A. Karyakin, E.E. Karyakina, H.-L. Schmidt. Electroanalysis (1999) 11 149.

-0.60 -0.55

-1

0

1

j, A

cm

-2

E, V

Catalysis of NAD+ reduction and NADH oxidation

0.1 mM NADH

0.1 mM NAD+

-0.60 -0.55 -0.50

-0.5

0.0

0.5

1.0

j, A

cm

-2

E, V

Equilibrium NAD+|NADH potential

A.A.Karyakin, Yu.N.Ivanova, E.E.Karyakina Electrochem. Commun. (2003) 5, 677-80

Direct enzyme bioelectrocatalysis

Protein electroactivity

Cytochrome C

S.R. Betso, M.H. Klapper, L.B. Anderson. J. Am. Chem. Soc. 94 (1972) 8197-204. M.R. Tarasevich, V.A. Bogdanovskaya. Bioelectrochem. Bioenerg. 3 (1976) 589-95.M.J. Eddowes, H.A.O. Hill. J. Chem. Soc. , Chem. Commun. (1977) 71P. Yeh, T. Kuwana. Chem. Lett. (1977) 1145-8Niki K, Yagi T, Inokuchi H, Kimura K. JACS 101 (1979) 3335-40.

Promoters for protein electroactivity

M.J. Eddowes, H.A.O. Hill. J. Chem. Soc. , Chem. Commun. (1977) 71P. Yeh, T. Kuwana. Chem. Lett. (1977) 1145-8

gold

ē

N N

ē

Berezin I. V., Bogdanovskaya V. A., Varfolomeev S.D., M.R. Tarasevich, A.I Yaropolov. Dokl.Akad.Nauk SSSR (Proc. Acad. Sci.) 240 (1978) 615-618

Direct bioelectrocatalysis

OHeHO Laccase22 244

Est = 1.2 V

Enzymes for direct bioelectrocatalysis

Iron-sulfur clusters HEM

PQQ

Others

A.I Yaropolov, V. Malovik, Varfolomeev S.D., Berezin I. V.Dokl.Akad.Nauk SSSR (Proc. Acad. Sci.) 249 (1979) 1399-401

Direct bioelectrocatalysis

OHeHOH Peroxidase222 222

A.I. Yaropolov, A.A. Karyakin, S.D. Varfolomeyev, I.V. Berezin. Bioelectrochem. Bioenerg. 12 (1984) 267-77

Direct bioelectrocatalysis

222 HeH eHydrogenas

BIOELECTROCATALYSIS by Th. roseopersicina hydrogenase

222 HeH

HeH 222

(1), (3) - H2 ; (2) - Ar (3) - without active enzyme

(Yaropolov A.I., Karyakin A.A., Varfolomeyev S.D., Berezin I.V. Bioelectrochem. & Bioenergetics 12 (1984) 267-277)

Equilibrium hydrogen potential (100% energy conversion)

Nernst’ equation for H H e2 2 2

How to involve oxidases in bioelectrocatalysis?

• surface pre-treatment;

• using of promoters;

• surface design by conducting polymers.

Fundamentals of direct bioelectrocatalysis

Investigations of enzyme redox centers

Redox switching of enzyme activity

Direct bioelectrocatalysis by

intact cells

Principal structure of bacterial cells

Inorganic ion reducing bacteria

Shewanella putrefaciens

Lactate as electron donorInsoluble Fe3+ as electron acceptor

Electroactivity of Shewanella putrefaciens

A – air exposed cellsB – air exposed with lactateC – no air, but at + 200 mVD – at +200 mV with lactate

Kim, B. H.; Ikeda, T.; Park, H. S.; Kim, H. J.; Hyun, M. S.; Kano, K.; Takagi, K.; Tatsumi, H. Biotechnology Techniques 1999, 13, 475-478.

Geobacter sulfurreducens on graphite electrode

Bond, D. R.; Lovley, D. R. Applied And Environmental Microbiology 2003, 69, 1548.

Advantages of bioelectrocatalysis:• a possibility for electrochemistry of complex organic

reactions;• high efficiency at room temperature and moderate

overvoltages;• achieve high specificity.

Disadvantages:• inherent instability,• large dimensions of biological catalysts.