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Yevgeni TomsovMy Dad

Bald DHT Chassis Cofactor Results Comb Vision

1 million people!


61 million people!


Suffer from baldness symptoms

Don't suffer

55%45%

N=470

Male pattern baldness by the age of 35

Practices


Effective Not effective

How effective are the solutions available?

7%

93%

N=470Practices


Hair tattoo

© Jill Carlson, flickr © Hagai Dekel, ynet

#trumpcat


SurgicalNon-surgical


Non-surgical treatment

Oraldirect27.5%

Topicalindirect

28.2%Other44.3%

Relevant Research, Inc.International Society of Hair Restoration Surgery2014




Topicalindirect




Oraldirect Side effects


Testosterone Dihydrotestosterone

5α-reductase

The cause


Dihydrotestosterone (DHT)

©http://gondronglagi.blogspot.co.il/2013/10/ingin-gondrong-lagi.html


SurgicalNon-surgical


NADH/NADPH NAD+/NADP+

3α-hydroxysteroid dehydrogenase

DHT 3α-Androstanediol

Human 3-alpha hydroxysteroid ... The Journal of Steroid Biochemistry and Molecular Biology 141, 135-143.


Bacillus subtilis: 15.4%

The most widespread bacteria on the scalp

Secretes

Roia, F.C.; Vanderwyk, R., R. W.. Journal of The Society of Cosmetic Chemists. 1969, 20, 113-134.


Yes No/don’t know/other

Would balding men use a product which includes genetically modified bacteria?

22%

78%

N=470Practices


NADH/NADPH NAD+/NADP+





For B. subtilis


NADH/ NAD+/



NADPH NADP+



F. Canonaco, et al. FEMS Microbiology Letters 204 (2001) 247-252.


F. Canonaco, et al. FEMS Microbiology Letters 204 (2001) 247-252.

NADPH

NADPH

NADPH

NADPH

NADPH

NADPH

NADPH


For BL21

For MG1655

Bald DHT Chassis Cofactor Results Comb Practices

For MG1655

For B. subtilis


Secretion Assay Principle

MIT OpenCourseWare

evrogen.com


Secretion with signal peptide aprE from B. subtilis

200

400

600

800

1000

1200

1400

1600

1800

2000

0 1 2 3 4 5 6

No

rmalized

flu

ore

scen

ce (

a.u

.)

Time (hour)

mCherry signal peptide+mCherry

BBa_K1674000


Activity assay principle

MIT OpenCourseWare

George H. Patterson et al. PNAS 2000;97:5203-5207

In silico model:


Activity of 3αHSD in E. coli BL21 lysate, DHT=40 µM

6000

8000

10000

12000

14000

16000

18000

30 300

Flu

ore

sc

en

ce

(a

.u.)

Time (min)

3αHSD no 3αHSD

BBa_K1674001

In silico model:


Reaction rate of NADPH consumption in different DHT concentrations

0.26

0.28

0.3

0.32

0.34

0.36

0.38

0.4

0 20 40 60 80

Reacti

on

Rate

(1

/se

c)

DHT (µM)

3αHSD no 3αHSD

BBa_K1674001

In silico model:


Cofactor assay principle

Extracellular Intracellular

MIT OpenCourseWare


Normalized NADPH fluorescence vs. Time for E. coli BL21 strains

BBa_K1674004

0

10000

20000

30000

40000

50000

60000

70000

0 4 8 12 16 20 24

Flu

ore

sc

en

ce

/OD

(a.u

.)

Time (hour)

BL21 wildtype BL21 with zwf

0

5000

10000

15000

20000

25000

30000

35000

Flu

ore

scen

ce/O

D (

a.u

.)

BL21 wildtype BL21 with zwf


Combined experiment principle

ExtracellularIntracellular

MIT OpenCourseWare


Reaction rate of consumption of NADPH from enriched supernatant

0.1

0.11

0.12

0.13

0.14

0.15

0.16

0.17

0.18

0.19

3αHSD no 3αHSD

Re

acti

on

Rat

e (

1/s

ec)

Combined experiment

BBa_K1674001 BBa_K1674004


?


Clean

Uniform applicationSafe

Reusable

The CombDesign


Optimal storage conditions:-18°C in 80% glycerol medium

Practices

Benefits of our solution


Purified enzyme solution vs.bacterial secretion

NADPH User experience

Achievements

New Application BioBricks Design

Modeling Collaboration Practices

Our Mentors:Dr. Roee AmitDr. Orna AtarNoa KatzInbal VakninLior LevyAlex Ereskovsky

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