1 2 3 4M

36

kDa

28

55

95130

5

Supplementary Fig. 1

Supplementary Fig. 1. Expression and purification of Bacillus sp. BRC1 Bdh in/from E. coli. Lane 1, total

cell lysates of E. coli BL21 harboring pET28a (control); lane 2, total cell lysates of E. coli BL21 harboring

pET-bhd; lane 3, soluble fraction; lane 4, insoluble fraction; lane 5, purified Bdh (arrow).

pH

2 4 6 8 10 12

Re

lati

ve A

cti

vity

(%

)

0

20

40

60

80

100

Temperature (oC)

20 30 40 50 60 70 80 90 100

Re

lati

ve A

cti

vity

(%

)

0

20

40

60

80

100

A

B

Supplementary Fig. 2

Supplementary Fig. 2. Effects of pH (A) and temperature (B) on Bdh activity toward the substrates D-2,3-BD

(closed circles) and acetoin (open circles).

D-2,3-BD (mM)

0 200 400 600 800 1000

Sp

eci

fic

Ac

tiv

ity

(U m

g-1)

0

10

20

30

40

50

Meso-2,3-BD (mM)

0 500 1000 1500 2000

Sp

eci

fic

Ac

tiv

ity

(U m

g-1)

0

10

20

30

40

50

Acetoin (mM)

0 10 20 30 40 50

Sp

eci

fic

Ac

tivit

y (U

mg-1

)

0

3

6

9

12

15

Diacetyl (mM)

0 10 20 30 40 50

Sp

eci

fic

Ac

tivit

y (U

mg-1

)

0

2

4

6

8

10

A

C

B

D

Supplementary Fig. 3

Supplementary Fig. 3. Enzyme kinetics of Bacillus sp. BRC1 Bdh. A, D-2,3-BD; B, Meso-2,3-BD; C,

Acetoin; D, Diacetyl

Rel

ativ

e ac

tivi

ty (

%)

0

20

40

60

80

100

120 BRC1/pWH BRC1/pWH-bdh

Xylose 5%

Xylose 5%

Xylose 20%

Xylose 20%

EFB hydro

lysate

EFB hydro

lysate

Supplementary Fig. 4

Supplementary Fig. 4. Enzyme activity of Bdh from recombinant Bacillus sp. BRC1 strains. Closed bars, D-

2,3-BD; open bars, acetoin.