Charakterisierung der reduktiven Aktivitäten der CO-Dehydrogenasen aus Oligotropha carboxidovorans und Streptomyces thermoautotrophicus

(1)

! "

#

! $

%! & & $ &%

!

(2)

! ' (

' ) & ! & * ( &

) + ,-& . ,//0

1 2 3 + -4& 5 ,//0

) 6+

) & ! & * ( ' ( 7-& 8

) & ! & 9 ! ' : ( 7,& 8

) & ! & 2 ! ' 1 ; << 7 " 8

) & ! & ' ) " ;

)!& ! & & ( = ' .

(3)

<$9 '1> #?@#< 9$<> <

" # -

"$" % & ' -

-&-&- ! $ " * ,

-&-&, ! $ " * 0

"$ ( ! # ! ) 0

-&,&- ! @ = *%! A

-&,&, ! @ " *%! B

-&,&C ! ( * *= 4

"$* + , % - . /

/ & & --

-&C&- --

-&C&, % -,

-&C&C -C

"$ 0 ! -0

"$1 % & # 2 / -D

"$3 2 & 4 // )) -A

"$5 . 2 & -E

% % ,/

$" ! % ,/

$ . 2 ! ,/

$*$ ! ,,

,&C&- 9 @ = ,,

,&C&, . ,C

,&C&C 9 . < . 79< 8 ,C

,&C&0 ,0

,&C&D 2 " ) ; ,0

(4)

<$9 '1> #?@#< 9$<> <<

$ $ 6 & // ,D

$1 // # 2 / ,D

,&D&- *%! % ,D

,&D&, ? " " ,D

,&D&C ? $ " ,A

,&D&0 ? $ " $ ,E

$3 // 2 // / 0 ,E

,&A&- " ,E

,&A&, ,B

,&A&C $ ,B

$5 % ,4

$7 // 8 ,4

$9 ( ( ! % C/

,&4&- F % . . . C/

,&4&, # . " 7#>?8%>. . C/

$" ! 6 ! / : ( ;6 :#< C-

$"" ) & CC

$" CC

,&-,&- < *%!

> CC

,&-,&, < *%! % C0

,&-,&C < *%! C0

$"* % : = ! C0

,&-C&- G %1 C0

,&-C&, . ! CD

$" +> 2 ?

CD

$"1 / : CA

(5)

<$9 '1> #?@#< 9$<> <<<

* #'( / # & CE

*$" % ( ( = ! @

CE

*$ ! C4

*$* A 0 0 0 & )

0C

*$ 2 & 0D

C&0&- ) " %? 0E

C&0&, > ; " %? 04

C&0&C 2 " %? D-

C&0&0 9, # %! " %? D,

C&0&D ? " @

* ! DD

C&0&D&- " %?

*%! DD

C&0&D&, " %? %

*%! DB

C&0&A ? @ [ >( *,]

*%! < " * A/

C&0&A&- 2 " F *% *%!

A0

C&0&A&, #>?% . . # " F *%

*%! AA

C&0&A&C *%*= <

*%! =

" @ E-

C&0&E 2 *= % ? *%

! > 2 . E0

C&0&B F % . . # *%!

" %? E4

(6)

<$9 '1> #?@#< 9$<> <

*$1 0 & B,

C&D&- ? $ " B,

C&D&, > ; $ %? BC

C&D&C 2 $ %?

*%! B0

C&D&0 ? $ @

[ >( *,] *%! B0

C&D&0&- $ %? *%! <

BD

C&D&0&, $ %? *%! <

% BE

C&D&0&C $ %? *%*= B4

*$3 0 & )B

4-

C&A&- ? $ *%! " $ 4-

C&A&, 2 $ %? 4C

C&A&C $ %? *%! 40

C&A&0 $ %? *%

*= 4E

99

$" 2 + 44

0&-&- # " *%! 44

0&-&, 2 ? " "

! *%! -//

0&-&C " %? *%!

G -/,

0&-&0 < *= % ? *%

! " F * -/D

0&-&D " %? *%

! -/E

0&-&A ? = " *%! " -/4

(7)

<$9 '1> #?@#< 9$<>

0&-&E 9 . ? " %?

H >( *,I% *%! --/

$ + 0 0 2 // / !

--,

0&,&- ? $ " *%! --C

0&,&, ? $ *%! --0

0&,&C $ %? *%

! --D

0&,&0 9 . < *%! $ +

J( K L --A

0&,&D $ %? H >( *,I% --E

0&,&A 9 . $ %? *%! --4

$* ! /( ' 2

0 0 +

-,/

1 . // ) -,C

3 // -,D

5 C 2 ! -,E

(8)

2K?@ $ #> #?@#< 9$<> <

& B 2

.

M M ;

> . %>. .

&

( 7 18

> ?

* *, 2 = 2 =

*%! 2 = %!

2 .

>

! )<) , A%! %. .

!(>* ! =

!$ ! =

!>(@ ! > (

@

#>? # . "

#N > = =% .

! % %!

#

( # %( %2

H, %,>I%@ # %> %@

% % 7! 8

>

9, G

9#)#> 0%7,%9 = 8%-%.. "

9< 9 . < .

<$1 ,%70%< . 8%C%70%$ . 8%D. %,9%

1 "

! 2

%G ( %( %2

( .

( ! ( . % %!

(9)

2K?@ $ #> #?@#< 9$<> <<

( (

()! ,%( %, 0%)

()(> -%( = . " %D%(

()1 ( .

( = ( = ( " (

$ !7)8 $ % %! 7,O%( . . 8

$ !7)89 $ % %! 7,O%( . . 8

"

$ ) . . $

& &

$

* >

*!0CA *. ! 0CA

) # ) % .

)(> )

)?2 ) .

. . (

? ? . . % =

> >

>!> $

1#(#! $ $ $P $P%1

F F '

F .

= ( = > "

F .

(10)

#<$'#<1 $ -

" #

2 = 7 *8 % &

# .

& $ Q "& & G

) " # ; . &

1 ( * & !

2 = ? 9

; " %*= &

* ?

" *, = & @ ?

? ' * " *, " =

* # % 2 %Q " 7

> -4B/8&

"$" % & '

! . *% 6 ! . 5 0&D4C

7# . ? . ,//-8& ! " <

*%# & !

;6 2 =

) % 9 & < $

*%> " (

>. 7/ /A / -D .. 8 * .

7(; & -44,8&

" $ " * ; " .. +

= . . .

( &

*% = ( 3

: "& & G 2

! 2 & > ; " ..

= . & # %

%. " "& &

7 & -44/8 .. 7)

,//C8 ; &

(11)

#<$'#<1 $ ,

! . # = . *

2 % # 3 .

> " & * " 2 = 7 *,8 =

!" !" 7 %@ 8 7(

> -4BC ( -4B48& = .

. > >

$ $ # " . 7(; -44,8&

! > " *%*=

= .

# " ( 2 . 7!

,//,8& ! *%!

$ > 7(

,///8&

* *= . "

) ( # #

& 6 *%*=

. . . ( G 79,8

7(; -44,8& > ( 7,//-8

" J9 L

( % .. *%*= 9, # .

& < J . L 9, J % % L%?

; 1 . ! & < 9

< G # 3

) G . ? 7> .

& ,//C8& ! ( C/R $ *%

*= G "

9,% &

"$"$" 0 2 !

*(D 7!>( -,,E #

7( & -44C 8 ' .

G * 2 % # 3 "

" & ! > " * *%

! 7( -4B,8& " "

(12)

#<$'#<1 $ C

*% *%

@ " 7( & -4BA8&

*%! G " BES

. " K

G . . " D/S ( " 7(

? -4B0 ? -4BD8& ! *%!

. # & !

# *%*=

. # " . DA- *%

75 " ( -4B4 ( -44/ (

& -44C 8&

* T 9_,* T , _DA- 7 =8 *_, T , 9^TT , _DA- 7 8

! # %*= DA-

> "

. > * 7 . ( -4BC ( &

-44/ ( & -44C 8& -AS *, *%*=

) . . 7 % 8

& ? . . % = 7? 8 ) . %

7)?28 > " = .

> & *,% =

? 3 $ !7)89F9^T & !

. G * ( .

# . 7( & -4BA 5 "

( -4BA8& G > 6

# *% DDB

*= -7 . (

-4BC8& G = .

' 9, # 3 *, %Q " & #

9 " 2 %<%9 ;

*= 9, 7> ( -44E8& !

# 9,%*=

(13)

#<$'#<1 $ 0

" . # " .

7> -4408&

"$"$ 0 2 !

. *% 9,% =

. ! 2

7 -44/8& ! *%!

* * " 2 %

# 3 " & " "

&

1 . " " 1 .

7 -44,8&

*% 9,% .

' $% ( " & !

*

G * 9, > = 7 -44,8&

*%! 7> C%) 8 . " @ *%*=

> . = # > & ! ?

6 ( % > . = %*= 7> ,%

) 8 = & ! # > C%" > ,%

) *,&% 2 .. & #

? 3 " " > ,%) > -%) & ! " > -%

) ? $, 9, 7? & -44E8&

"$ ( ! # ! )

! 2 7 & -%-8 *%!

= . " "

9 ! "

# 7! & -444 ! & ,//,8& #

9 + -8 ( . 7'% 8

@ ,8 . 7(% 8 !

C8 # . 7>% 8 " >%@

7! & -444 & ,///8&

(14)

#<$'#<1 $ D

! '% 7 ='8 ( % . %) " & <

( BB E ! & $ (

2 ( . % %! 7( !8& !

( ! ) . %! )

) 7( & -44C 8& ! (

# ( ! &

&&$" " @

! > ! *%! & !

. 7 =(8 # %

> %) 7 =>8 6 ( . 7 ='8

&

! (% 7 =(8 . "

( C/&, ! & > %

2 !& ? =( $

" " ! 7 ,///8& ! < !%

=( =' => =(

2 " !% ;

7 -444 ,///8& ! >% 7 =>8

( -E B ! " H, %,>I% .. #>?%>.

1 .< 1 .<< " 7(

(15)

#<$'#<1 $ A

-44C ,///8& ! %>%@ 1 . <

2 " '% >% %>%@

1 .<< 2 >% (% &

! # 9 *%!

& ! # 6 *%*=

H >( *,I% ( %2 H, %,>I%@ 1 . <

1 . << ! . " .

7! -444 ( & ,///8& ! # ;

%. # " . = (

1 1 ! . % . 7! )<)8 ) ,%70%

< . 8%C70% . 8D%. %,9%1 " % 7<$18F-%( = %

. " %D%( 7()(>8 7( -4B, 2 &

-4B4 ( -44C 8& < > 7*,8

# *, U > . =

G . = " < # " 7*

-44E8&

< ? =

*%! $ " $

" & ! % " ( % "

# %! $ %? 7( & -44C 8& < "

" $ $ !7)8T9^T !9,

# %! *%! & ! ( %@ *%

! $ %? & *

# %! $ %? 7( -44C 8&

N 9 . = ) .

> *%! " 7( &

-44C 8&

"$$" . / '

! @ > *%*= *%

! ( . 7 ='8 & <

" "

( %9 = @ *%

! ( ( & !

(16)

#<$'#<1 $ E

?; . % %>. . 7#N >8 7

,//C8 ?; . 7! ,//,8 &

! ( *= " T < ! %

> ( ! . ) *= %

.. & 3 ' 9 = % ..

72 8 " & *= % .. 7?; . 8

> & ! " ( %*%'

( &

&&$" ' ; < 2 ; < . /

; && $

<

! > = 7 8 " *%! 7 8

" " @ & !

& ! @

H >( *_,I% ( %%

' & ( 7T <8 7 = @ 8 ( 7T< 8 7 "

@ 8 .. # .. ( !% &

7<8 7 = " @ 8

.. γ> CBB & ! ( . *= ..

3 *% 9 = .. & EAC

= 7* ,8 ( %< %) "

*= .. G J .L

7 ! ,//,8& ! @ . "

M&

(17)

#<$'#<1 $ B

< = @ % *= T<

> CBB '% & (

>% %> "

7 & -%, 8& ) ( %>

( %%>% --CR %>% %>%

-DAR 7! & ,//,8&

! 3 " @ . "

7 V? ^CBB> ?8 *%! & ! CBB

>γ % @ & @

,0/ EAC G " " *%

' ( & *ε, EAC " '

. ' ( . ; & EAC

G " > CB4 J .L7! & ,//,8&

"$$ . / 2

! ? *%! * 9, !

= "

% & ? *%!

( " 7( 7< 88 & !

*= " T< 7 ,//C8&

? 7 * 9, ! 8 > =

" *%! " "

@ 7! & ,//,8& " @

! " " ( (

C E/ M 0 ,C M 7?; . 8 7 & ,//C8 " &

C E0 M C 4C M 72 8 7 & -%, 8 7! & ,//,8& !

*%' ( ? . - E/ M

3 , -A M& < 2 .

*= .. " ( - ED M& ! 9 = .. " (

, /C M& < ?; . " @

3 *= .. " 9 = .. 7 & ,//C8& ! %>

;6 " ( , ,E M , C, M

72 8 " & , ,4 M , CD M 7?; . 8& !

" %> " >_λ & ! %*

(18)

#<$'#<1 $ 4

EAC ( C -0 M C ,4 M& ! ( %%>% %G

--, -,, >% %>%G -DA -A/ 7! & ,//,8&

> ?; . 2 "

" @ > 7 * 9,8 ?

7! & ,//, & ,//C8&

G * 9, = @ *%

! " ! = ..

2 7( > < > << !8 "

*%! & !

*%! *(%-,F.5?!(>'

@ " ! " & !

F % . .

H, ,>I%@ ! . 7 ,//-8& <

* 9, @ " & <

# " ; *%*= #

= 2 " &

; . . . "

@ *%!

> 2 . & ! @ *%

! ',( *C%>

! %> 7',8 ( *= % ..

7 ,//-8&

"$$* % ! / '

*%! *%*= " *, " ) "

# 72 .& -&,&-8& ! " # *%*=

( " & # H >( *,I% "

! *%*= ; 72 .& -&,&,8& #

> @

?; . 2 *%(

@ & 5 9 !

> @ < %

( ? *%*=

7 & -%C87! ,//,8& < ? %

(19)

#<$'#<1 $ -/

*%! 1 %

K " & # . ( * "

W%> %' 3 9 = ..

& ! *% * % %

%> 3 *%' ( & !

# . @ " "

( %' &

&&$" * 4 ( ! + / ! / '

!

! ? " < * =

H >( 7X*8*9I%@ " >% 3 *%'

( & < K " . 3 *%

*& ! 1 %@ ( 7 <8 " ( 7< 8 " &

! 1 ; *, & ! 3 *%'

9_,* " & ( #

# . " " = 7! & ,//,8&

*

9T

*, 9T ^T 9,*

, %

( >

>

*

*9

>

7 <8 7<8

( >

>

*

7< 8 ^7<8 >

*

( >

>

*

*9

7< 8 7<8 >

*= @

K "

? " @

(20)

#<$'#<1 $ --

< 1 %K " (

Y ; @ Y & # *ε,

%EAC > ( 7T <8 @ &

; @ ( *ε,% @

% > ( 7T <8

? & %>

% * # " (

& 6 ; > % *

# . ; 3 9 = % " &

*= .. *,& < K "

% * *% %>

% *= 7< 8 & #

*, " > 6 >

@ " # " 9 = % " & *= .. (

9,* & ! ( 7< 8 #

%>%@ >% " " ( 7 <8

= & ! G # ! (%

= # " . 7! & ,//,8&

"$* + , % - . / /

& &

"$*$"

! *%! 7( > -4BC8&

< D ( " 9 *%! %

@ -/ - 4- ? 7> $^%8 .

# " 7 ,///8& < " *%!

" % *%! #>?% F %

>. & < #>?%>. % *%! -A

2 ! %( 7T 8%>

> 1 .%<%H, %,>I%@ 1 .%<<%H, %

,>I%@ *%! 7 ,///8&

! F %>. % *%! "

" # " & C// A// &

(21)

#<$'#<1 $ -,

2 . " %< %%

> %' " @ % *%

! @ *%

' 7 & -%0 87! & ,//,8& ! %<

% 7T<<8 =

& ! ( 7 <8 @ *= ..

7. , ! .. ( !8 7! ,//,8& ! ?

% *%!

N %! > %'

%< &

&&$" . /

; < !

/ , % - ; <; && <

7 8 ! ( %@ % # " #

( ! *= % .. 7! ,//,8& 7 8 ! $%

% % %>

9 = .. ( 7! & ,//,8& ! @ .

" M$

"$*$ !

< = *%! %

# " & < * >

.% % # .

" & ! %

7 8

Glu 763

Cys 388 1,81

1,71 2,38

2,47 3,49 1,81

7 8

(22)

#<$'#<1 $ -C

< *%*= 7! & ,//,8& < " " %

*%! " @ %

% *%! & ! (

( % % *%!

*= " T< T< & ! 2 % %

*%! " < % ..

H >( *,8% 7 & -%0 8& G % %

%> 9 = .. ( $%

% & ! % % .. >

. 7! & ,//,8&

"$*$* !

! ' % = *%! " @

& # . = #>?%>

% # " " ! " & #

" Z > # " 7# ,//C8&

&&$" 1 +> !

. / ; # *<

,% %> @ -%

3 9 = .. ( & # %

3 9 = %* (

%>& ! ! " ! .. 7# .. 8 7#

,//C8&

(23)

#<$'#<1 $ -0

! 2 *%!

& ! > % # " 7 -%D8 - C

M " @ &

" ( %> 9 = %* & !

-% %> ,%

3 9 = .. ( 7# ,//C8&

"$ 0 !

< " " > "

*%*= $ % # % *%! & $

*%! 7$ % *%! 8 9

. %

" " 7'

,//,8&

! $ % *%! ? *

G *, " # " ) & <

$ % *%!

2 . = $ % # " % %

> 7? 2 -44A ? ,///8&

& "

. .. J9 L ; "

" $ % *%! "

& *%! <

*%*= #

) . " 9 7> &

,//-8& ! *%! << & ! *=

* $ !)^T%? " $ !)9 T 9^T .. 1

*, 2 & ! 2 *%

! << & ; -&A M

7! & ,//-8& ! # " 9

" "

H0 %0>I%@ 7 !8 .

& ! ! " H0 %0>I%@ H

OI " H$ %0 %D>I%@ H OI& !

(24)

#<$'#<1 $ -D

( O 9

) . & ! $ > %

%> ' 7! & ,//-8&

*%*= H$ %0 %D>I%@ &

"$1 % & # 2 /

( 7( 8 -4C/ " >. "

? > = . 7 -4C/8& @

# " 7 C/8 ( & < ( " (

G 7G 8 " & ! 7 . 8 .

79 ,//,8 G%> %

( %> % 75 & -44A8& ( G

; ) # " K &

( %# " " ? > %

> % 2 %2 79 -44A8& ( ( "

" + -8 # ( % # " ,8

( # % ( % # " & ! (

$ 2 . = ( #

> # %( %2 7 ( 8 "

79 ? -44A8&

> @

( % # " ..

7 & -%A8 79 ,//,8+ -8 N %*= + ' ^<*>7*98%

@ & ' ) %2 & #

. # " 9 = & ,8 *= +

> %*= % '(* ^<*,7>% 8% @ &

# > %*= $ %? ; "

.. & > 1 > " &

# . > & C8 *= + ! %

> = %? 7!(>*8 ',( ^<*7N8% @ &

( %@ .. " ) %2 &

! 7N8 " ' ; > 7 !(>*%? 8

7 $ %? 8 > 7 %! 8

9 = FG %( 7 %*= 8 & (

(25)

#<$'#<1 $ -A

.. 1 >

9 = &

; < ; < ; <

&&$" 3 . / D ' ; < )

' ; < / ) ' + ; % <; <

8 / ;4 <

"$3 2 & 4 // ))

" @ '

# '

7 ! . .. "& $C%8 7G

-44D8& ! "

*,&

! " J L 7 8 '

K 2 & !

# % " % " %2 . = & <

" ( . = '

" "& & J " L ( %1 . # " #

7 ) "" -4ED8&

( 7-4A48 " %( . = *

" < " & ! "

< [ ; 1 . *%! 7,D/%

C// . 8 ; 7 9 -4AB8&

7>% 8 7> % 8 7%*98

> ( *

> >

*9

> ( >%

> *

*

(

> >

* *%

(26)

#<$'#<1 $ -E

$ " # " " " &

" ! 7$,8 > $ & !

" ? " 7! 1 -4B-8+

< " " " ( % ) "

& ! " %1 = " ( .

2 " ( " @

# " 7> ,//,8&

< ' " < " >

# . = ) . 72

9 -4C48& " # 7 '8 % %*=

# " %

72 . =%< 8 > " G " " &

" 2 . . " <

& " 2 . = 2 . .

7) -44E8&

"$5 . 2 &

! = # *%!

<

# " ? " & <

< " *%!

' > " " & # @

? " *%! " &

? H >( *,I%@ " &

" >

*%! & #

? & ? *, " * *%!

0_* E * 4^E 04_*E 0

0_* 3 5 4^E 0 4 E 04_*

E

E E

0_* 7 9 4^E * 04_*

(27)

#<$'#<1 $ -B

& ! " " *, & ! $ %

( ;6 " "

" 2 . % $ %? 79 &

-4B48& $ 9 % # " & (

" 7-44/8 "

*%! ' $ " $ "

" & ! " $ 9 *%

! & >

" $ $ ? *%

! &

@ >

*%! " F " G %

# " & !

> # " H >( *,I%

*%*= % ?

? . & # ( 2 . 7 8

*= *%!

? ( ( ?

. & < >

# " $ 7> &

-44E8 \

. # %! %. # %

! ? &

@ " >

2

? 2 " &

< " *= %

? " *=

# ? ; & !

# %! * 9,& !

( " *= & & %

% %< %

? " & ! 9

(28)

#<$'#<1 $ -4

# ? ? "

% ? " ; &

! *%!

% .

& 5 " *%

! "

. ; %. # "

" 9

*%! % ; &

( 2 "

% G *%! # "

&

(29)

( 1#?< ' (#19*!#$ ,/

% %

$" ! %

!

*(D 7( > -4EB8

7 & -44/8 & !

> ' & *%!

*(D%-,F.5?!(>' 7,//-8

& ! *%!

' * " = & *(D%-,F.5?!(>'

) .5?!(>' 2 *%

! %> ( .5?!,-D &

$ . 2 !

! @ "

" ( & < ( > 7-4BC8

( 2 " >.

7<<<8$90% , ; $ ,9)*0 29,)*0 " &

@ 9 $ ; - DS % (

" &

% / /

#

$ ,9)*0 = ,9,* 29,)*0

$90

( >*0 = E9,*

, = ,9,* 7<<<8$90%

>. ; 1>, 7-/=8

3 &

.9%G

- B/

/ ED - D/

/ 0/

/ /0 / /C - ,D -/// //

E //

/ 4/

- D/

/ ,/

/ /, / /,0 4 - //

-/// //

E //

(30)

( 1#?< ' (#19*!#$ ,-

( / > ;" )! @ 2<

# ( 7 8

@ >*0= E9,* / -/

( ,= 09,* / /A

9C *C / A/

, / 0/

,= ,9,* / /,

$ ,= A9,* / /0

$ ,> *C / /,

$ ,( *0= ,9,* - B/

3 & 7 8 -/// //

.9%G E //

*% . G

. +

* S 7 F 8 D/ DC

*, S 7 F 8 D -/

' S 7 F 8 0D CE

! 2 -// % 0// # %2 7,D/ ] -/// 8

G P 1 . ? 7 .

8& ! < C/R

ADR &

! ;6 @ C/ D/

7 ( 8 ,//%C//

(31)

( 1#?< ' (#19*!#$ ,,

. & ! ADR 7 8

C/R 7 8 2

BDS 7 F 8 ' -/S 7 F 8 * DS *,

, ^%- & $ . *!0CA / , ] / C

*,% & ! 2

? @ -D/ 0D/ ^%-

; & G .9%G

@ D $ $ *9 " & ,/S 7 F 8 9C)*07'; G 8

E / &

! G

( . ! . 0CA

>. . 7$ . << ) 8

& ! 6

. ? & $ # .

! & / 4 7 8 " & 0 / 7 8

2 9 ! " 7@ , .

@0- ) 9 ' 8 -A&4D/ & ! @ .

> 7%-4,R 8 " ]

B/R &

$*$ !

$*$" 4 . '

@ 72 .& ,&,8 D/ ( 29,)*0F$ *9%) .9 E ,

7, ) ^%- @ 8 9 9

& ! > . 2 !$ <

# ; ) 7)(> 8 7# " / D (8

" & ! @ A E )

) %@ 70/ & > >. (

> 8& @ # " @ @

? = C/ " 7B&/// 0R 1%

-,0 2 # 8& ! K " ? = & !

,% " 7CB&/// 0R 1 1 0D&4D

1%-/AD 2 # 8 @

(32)

( 1#?< ' (#19*!#$ ,C

@ & K

. &

$*$ ! ! / (

! . 9 N2 D/%>

>* ? # C/Q 7 ) 8 1

-// & !

) Z21 ¹⁽ ) -//% 7

) 8 & ( D ^%-

. D/ ( 29,)*0F$ *9%) .9 E ,

3 > D/ ( 29,)*0F$ *9%) .9 E , "

# ) . & 6 #

) $ % 7/ - ( $ D/

( 29,)*0F$ *9%) .9 E ,8& -/ ^%-

[ -/ & G

) ( . ,B/

& % %>%

. 0D/ DD/ " & ! *%! %

" 2 .& ,&D&- &

$*$* 4 ( & ! / ( ;4 <

! 1 . " . . < .

) " . ( & !

) " ( = *

) & ! > " "

"& & ; . G

& > > " 1

" &

! 9 . < . 79< 8 N2 D/%

> >* ? # -D<>* 7 ) 8

7 -// 8 & !

) Z21 ¹⁽ # & -//% &

(33)

( 1#?< ' (#19*!#$ ,0

! ) . > - , (

7$908,>*0 7 ) 8 " & $ . " . )

@ 7B&/// 0 1%-,0 2 # 8

) > . . / BD ( 7$908,>*0

D/ ( 29,)*0F$ *9 .9 E , 3 > & $ )

-,/ / BD ( 7$908,>*0 D/ ( 29,)*0F$ *9 .9 E , &

) / BD ( / (

7$908,>*0 & 6 -/ ^%-

-/ & G )

) ( .

,B/ & % >%

. 0D/ DD/ " & *%! %

2 .& ,&D&- &

$*$ : )

. *%! % > " "

" 72 .& ,&C&D8 "

> . = %,D 7 ) 8

D/ ( 29,)*0F$ *9%) .9 E , & ! # "

3 & "

# " ]B/R &

! # " ) ; 7 , D 8

)!-/%> 7> . = %,D )

8& . ) 3 >

, D ) ; C D )

&

$*$1 @ 2 6 >

! ) ; 7 7D/

8 ( B/D/ G 8 ' - D

? # " & ! (

" C/ ! 7! )( C/ G 8&

(34)

( 1#?< ' (#19*!#$ ,D

@ 2 " 7 C D 8 @ %

2 " 7( . ( " C/ ! " )

> ? 8 " &

$ $ 6 & //

! ) " #= )

( " %( " 7-4DC8&

@ ) ) ; "& &

# .

7-4EA8 ( &

$1 // # 2 /

$1$" //

! *%! % . ?

,%70%< . 8%C%70% . 8%D%. %,9% " 7<$18 " -%70%

< . 8%C%. %D%70% . 8% " 7 " 8 72

& -4B48&

$1$ // + 2 2 // /

? "+

$ $C -/ (

$ D (

*%! / D%,

D/ ( 29,)*0F$ *9%) .9 A D / ED

! *%! ) G 7B

8 .. & 6 G

> . (

9 & ! $ $C%'; ! ;

G ? " 9 9 %>. "

& $ D A/R 9

(35)

( 1#?< ' (#19*!#$ ,A

9 %>. " / , ) ! .. "

" & " &

7-4B08 " ( >

7-4A/872 .& ,&A&,8 " ! 1 7-4B-8 72 .& ,&A&-8

&

6 ? " C/ A/R

& ! " "

# % 7 -8 7 /8 (

" %? & ? "

" ) " &

$1$* // + 0 2 // /

+ 2F

2$*, -/ (

$ -/ (

*%! / D%,

D/ ( 29,)*0F$ *9%) .9 E , / ED

*%! ) G 7B 8

.. 6

9 & !

$ % 2$*,%';

9 9 %>. " G & $

D A/R 9 9 %>. "

/ , ) ! .. "

" & " &

7-4B08 " ( > 7-4A/872 .& ,&A&,8

$ 9 ( 7-4AA8 72 .& ,&A&C8 &

6 ? " A/ A/R

& ! " " # %

7 -8 7 /8 (

$ %? & ? "

" ) $ &

(36)

( 1#?< ' (#19*!#$ ,E

$1$ // + 0 2 0

+ 2F

2$*C B/ (

$ - D (

( / 4 (

*%! / D%,

D/ ( 29,)*0F$ *9%) .9 E , / ED

! *%! ) G 7B

8 .. & ! G

9 & 2$*C%'; (

$ %! ;

G ? " 9

9 %>. " & $ " D A/

9 9 %>. " / , )

! .. " $ % " & $ & $

9 ( 7-4AA8 72 .& ,&A&C8 $

7-4ED8 U ( 7-4B08 " &

6 ? " A/ A/R

$ & ! " " # 7 -8%

7 /8 $ ( $

$ %? & ? " "

) $ &

$3 // 2 // / 0

$3$" // 2

@ " %2 " ! 1

7-4B-8 ( & @ 1 " )

@ %) " 7( . ( " C/ !

" ) > ? 8 " & ! #

" & ! " . . # "

7 ^TC% " 8 '; & # ) / D

[ - %) F9 7/ - ( F9 .9 , /8 / D C

(37)

( 1#?< ' (#19*!#$ ,B

7/ , ( C / /- ( 9 ? 8 " &

6 #= % ; '; -D

< ? . 2 > X -

) 0DB 7$ . << ) 8

& @ # " #=

# $ $C > 7/ ] -// W "^%-8 &

+

0DBX , E,^%0&2 " " 7W &D// W^%-8 T 0 00^%C

$3$ // // /

! 2 " 7-4B08

" ( > 7-4A/8 & #

) / - [ - 4 3 & - $ %)

7,S8 F 8 / D $ %$ . 7/ //DS F 8 / D $ %9 .

7/ , ( $ %9 . / -,D $ $ *98 " & ! " A/

C/R ! & 6 #=

'; 2 > X - )

AC/ & @ # #=

# $90 > 7/ ] -/ W "^%-8 &

+

AC/ X %/ //,/0&2 " %$ 7W &-// W^%-8 T / -BE4A

$3$* // 0

! $ 9 ( 7-4AA8 &

# ) - - ? " ;

1 / BS > 7 F 8 D $ #

DS α%$ . 7 F 8 D $ # " & 6

#= '; -D <

? . 2 > X - ) DD/

& @ # $ #=

# 2$*, > 7/ ] 0 W "^%-8 &

+

DD/ X / /-AA0&2 " $ 7W &-// W^%-8 T / C,ACA

(38)

( 1#?< ' (#19*!#$ ,4

$5 %

! # ( 2 . ( @ > (

< 1

: 7 <1:28 < % . %) %

( . 7 ) 3 )Q, 1 ) < F

# G 8 &

! 2 . *%!

3 # " ;

& ! ( .

>. . --// 7) # K -/ ' .

2 . . ,C0 8 + & 0 ) - C

& # / -A W( 2 . ; &

! ) 2 . "

# 7^ B W( *%! 8& ! 2 .

) &

$7 // 8

! *%! 1 =

( . " " = "

0D/ 7( -4B,8& < #.. %? 6

A// µ *%! 7/ D - D ^%- 29,)*0F$ *9%)

.9 E ,8 A/ µ D/S 7 F 8 1 ,/ 0R

& 6 " C0/ µ , 0 ( 2,9)*0

) " . D -C&/// " & K

@ 2 ! . 0D/

& ! 2 " 9 ? =%

! "%#= " ε0D/ X -/ D (^%- ^%- 7G

-4ED8&

(39)

( 1#?< ' (#19*!#$ C/

$9 ( ( ! %

$9$" G ? & ( ( (

F % . . ) ; @ %

>. . 7 40- 2 # 8

. ) ? " ,

,// ^%- ,C/ BD/ " &

! >. *%! ! % 9,% " &

*%? " %? Q "

> . > &

$9$ # ( 2;# +< ( (

! # . "%>. 7#>?%>. 8

#(N%#>?%>. 72 8 & ! >.

9 %2 7#>? 4// *= < # 8

1 . %2 7<1 D/C *= < # 8

1 . A 2 -,/ 2 ; & ! >.

( -/ " & ,// G " & !

#>?%>. [ " & ! !

> G<$%#)? 7 8 " & !

( 3 " ( "

2 ( ! . . " %? ". &

! % +

ν+ ( 3 " 9"

+ ( 1

*%! 7-/ -, ^%-8 D/ ( 9#)#>F$ *9%) .9 E ,

= 7D% #

-//S 9 8 0 ( ! " & *= "

) [ .

E- 00B0 &ν 7 T / ,AE8 X

(40)

( 1#?< ' (#19*!#$ C-

9 %>. " #>?%?; 6

&

$" ! 6 ! / : ( ;6 :#<

) % . 7) #8

) . > ' 7-4E/8

& ! ! 1 -,/ 9 = -,D

= - D 1& # ,/ > 1

2 " -/ ) 7,,D W 8 " &

4 : F

C> F

F %'; + C/S 7 F 8

$ $_%( / BS 7 F 8

1 . + / 4B/ ( 1 F9 7.9 B B8

> . + / C,D ( 1 F9 7.9 A B8

' . + / /D/ ( 1 / CB0 ( 7.9 B D8

/ -S 7 F 8 >!> 7 8

! " ! # . ' .

# . %2 " &

H & " 6( ! / )B 6 ! /

1 >

E DS -CS DS E DS

F 7 8 4 // -A 0/ - AE , D/

1 . 7 8 -B // -B // % %

> . 7 8 % % D // D //

3 7 8 4 // C A/ , 0D , 0D

$ ,>*C7 8 D/ // D/ // % %

>!> 7-/S F 8`⁸ 7 8 % / CA % / -/

1#(#! 7W 8 4 // 4 // D // D //

. = 7-/S F 8 7W 8 4/ // 4/ // D/ // D/ //

`⁸

(41)

( 1#?< ' (#19*!#$ C,

! ) ) " -// W

= -// W .. -F-/

/ ,DS 7 F 8 . -F-/ 0/S 7 F 8 > "

9 %>. " " 1 &

) )

7D/ -// W 8 @ . > 7-/ -D W 8

,S 7 F 8 >!> " D G &

! # . > -D >

0D 1 7$ " + ! -///F,// ! 9 8&

> . % #

&

@ ( )

> -0 40 ! 7)

8 & ! ( %

J9 ( G 2 # . L&

$ # . )

) ?%,D/ A/

9 " 7D/ . ( 8 = & 6

" #

# ; &

8 & > F

?%,D/ D//

1 & # 0D0

# 4,

9,* 0D0

# ) & > F

9,* 0//

( D//

# -//

! ! ) ! < 7(G %

# 8& ! < ) "

(42)

( 1#?< ' (#19*!#$ CC

( 9 > ? ')

* ! > 7 ( > 8 &

$"" ) &

! *%! %

) 72 .& ,&-/8 ,%70%< . 8%C%70%$ . 8%,9%

1 " 7<$18 -%( = . " %( 7()(>8

&

! 1 " C/ > % 9 2

* 6 > &

< ? . ? -

? <$1 &

$"

$" $" !

// ! & ! = )

! *%! 7( > -4BC

,///8 ; > & / 0

" = ,D/ ( 2% %'; 7 29,)*0F$ *9%

) .9 E ,8 " 4 A = *%! 7C ^%-

29,)*0F$ *9%) .9 E ,8 ? . &

/ E 3 ? " . #

@ 7C/ DD// 8 ( C/ !

" 7@ %2 " ( . ) > ? 8

& # ? 7> $^%8 >:? *> ? " "

# %? 7 > $8 " 7G -4B-8& ! # 7/ D

8 2 >:? *> ? " 7/ D 8 -/

& ! > $ . 0A/ &

? " " # / D -/ (

2 $ D/ ( 29,)*0F$ *9%) .9 E , & ! Q "

. 0A/ # 2> $ 7/%,/ µ(

29,)*0F$ *9%) .9 E ,8 & ! *%

! ( "

(43)

( 1#?< ' (#19*!#$ C0

> . = %,D 7)!%-/ ) 8

" & > 6 *%*= 72 .&

,&D&-8 " % 72 .& ,&D&,8 $ % 72 .& ,&D&C8 $ %? 72 .&

,&D&08 &

I+ + 2 G 7-4B-)F

7$*C8C = 49,* -/

ADS 9$*C ,/

3 & -D/

$" $ ! !

! = *%! % 7!

,//,8& ! = 1 " - ) 29,)*0F$ *9%

) .9 E , 2 " 7/ -

- , (8 % " & $ -D <

? . ? " 1

*%*= 72 .& ,&D&-8 " % 72 .& ,&D&,8 $ % 72 .& ,&D&C8 $ %

? 72 .& ,&D&08 &

$" $* !

! = 1 " - *%! -

29,)*0F$ *9%) 7D/ ( .9 E ,8& ! 1 " $ %

! 9, * " & = 2% %'; 7-/ C/ (8

" 1 " D/R & ! "

. " 3 * → <$1F()(>%1

72 .& ,&D&-8&

$"* % : = !

$"*$" A & H !

( G %1 ? %

G % 7 ( 8 !

(44)

( 1#?< ' (#19*!#$ CD

'; >. " & ! G

1 . 0/R &

+ 2F

9 . + , 29,)*0F$ *9%) 7D/ ( .9 E ,8 , *%

! D ( -D ( " &

@ " + / , 2*9 7,/S8

! 6 -/ 9 . & ! ?

@ -/ ( ! &

$"*$ : ! / ( ! // ! ))

! 2 " ! " %

? . & ! )

. 7( ) 4///U . (

$ 8 JG %! L7( 4/C 8 . " &

> 9 Q 7, D 8 ( -C N 7- B 8& !

1 . * 0/R < [ ! B/R

,//R & 1 9 ! C/ ^%-

" & ! > 7'<$>#<>%'AD-,8

" & @ # > " #

> > 7/ ] ,// 8 &

2 +

) 7 8 X / -/BCD&2 " > 7 &D/ W^%-8 T / /C0,B

$" +> 2 ?

! 2 " F *% *%!

! . J 1 . L& 2

A W # " 7,0 ^%- D/ ( 9#)#>F$ *9%) .9 E ,8

! 7-B ! ! 8

, A W 2 . 7! -4448&

(45)

( 1#?< ' (#19*!#$ CA

! > ! ,/ C/%

! %'; 7-S 7 F 8 1 8 A/R & 6

! G . D -/ 4AS 7 F 8

# G . !

$ -C/R 9 " &

@ ( )) F

/ B, ( 29,)*0F2*9 / B, ( $ 9,)*0F$ *9 B, // ( 9#)#>F$ *9 .9 E ,

CS 7 F 8 ,%( %, 0%) 7()!8

3 &

! %2 7<G 2< . !

' 8 E// W 2 .

? > ; 7( ? ?

? 2 8 & ! ! 1 . (

A W " . *% *%! C W

2 . %

2 & ! 2 0R &

$"1 / :

! ?

7! 8 ( 7! 8 > 7! 8 7( 8

) 7 8 > 79 8

? 4ES " & !

' 79; 8 ? 7( F' 8 " &

< ? + * 44 CS *, 44 44S 44 DS

44 4/S 9 44 DS > 44 4/S&

(46)

#N)#?<(#$1# #? # $<>># CE

* #'( / # &

*$" % ( ( = ! @

! 2 " *% .

" ' %) = G

*!0CA - D ,& !

& # G

F ( F

*%! # "

& # G . *%

! )

" =.

) ) 7 & C%- 8&

G ' %)

7 & 0B 8 &

G

1 . 7C/R 8 2 [ "

*!0CA -/ -C 7 & C%-8& ! ; *%*=

( =. G . 7> -44A8&

! = .. " "

=. ) * &

"

*%! %>. " . "

&

(47)

#N)#?<(#$1# #? # $<>># CB

&&$* " ( A ! /

/ 1 8 / ; <$

/ & 8 & ; "< ) & ; <

! ( / ! 8

! ! . A ! / (

(

, ^%- -/S 7 F 8 * DS 7 F 8

*, BDS 7 F 8 ' *% . " 72 .& ,&,8& 7 8+ ! G

. ! 0CA & 7 8+ ! .

G .

72 .& ,&C&-8& ! .

) # 72 .& ,&-/8& -+ ,/ W > 79

( G ) 8 ( . + 1

7AA4 ! 8 700/ ! 8 2 7,C, ! 8 ' %! 7-0/ ! 8

? 7AE ! 8 , D+ D/ W . @ E,

4A -,/ -0A > . & 7 -8+ !

) 72 .& ,&-/8 7 ,8+ !

) 72 .& ,&--8&

(48)

#N)#?<(#$1# #? # $<>># C4

*$ !

*%! @

" %

. . < . 79< 8 &

! " *%!

[ " # & ! . .

*%!

& # = . *%

! &

*%! 0/ 7 8 *%

. @ " & $ @

@ " ? = "

. % ( 72 .& ,&C&-8& ! *%

! 7.<+ 0 E8 7( -4B48

% . & !

. . >

C/Q " 7 & C%, 2 .& ,&C&,8& $ )

) 6 G

> Z3 . 7D/ ( 29,)*0F$ *9%) .9 E ,8

& ) $ %

7/ - (8 & ! # *%! & / 0 (

$ & ! *%! . &

@ *%! %

> -D<>* 79< 8 > 7 & C%C 2 .& ,&C&C8&

$ ) )

6 G > & )

9 %

> & ! # *%! & / 0D (

7$908,>*0 7 & C%C8& . 0D/ 7( 6

8 DD/ 7( 6 >% 8

*%*= " .

(49)

#N)#?<(#$1# #? # $<>># 0/

72 .& ,&C&D8 " & @ #

# " > . = %,D 72 .& ,&C&08&

&&$* ! / ( ! ( / !

8 ! * J

! . D/ ( 29,)*0F$ *9%) .9 E , 3

> C/Q%> 7-// 8 & ! # )

>. ,// D/ ( 29_,)*₀F$ *9%) .9 E ,& ! )

$ % 7- D 8 / - ( $ D/ ( 29_,)*₀F$ *9%) .9

E , & ! ) -/ ^%- >.

,/ ^%-& ! ,/ & ! )

( . ,B/ & ! ( %

>% . 0D/ " & DD/ & ! *%! %

" . 2 .& ,&D&- &

< *%! *%

= & ! *%!

= "

2 @ "

? . & ! . *%

! =

" &

# 7 8

/ ,D/ D// ED/ -/// -,D/ -D// -ED/

/ D//

-///

-D//

,///

,D//

/ -//

,//

C//

0//

D//

/ D/

-//

-D/

,//

,D/

C//

/ ,/

0/

A/

B/

-//

/ -/

,/

C/

0/

D/

.,B/78 .0D/78 .DD/78 $7S8

*%*=7%- 8

(50)

#N)#?<(#$1# #? # $<>># 0-

&&$* * 4 ( & ! / ( ! "1

! ) . % . *%! ]

7$908,>*0'; " # " - , (

" & $ " K / BD ( 7$908,>*0

D/ ( 29,)*0F$ *9 .9 E , 3 > 7-// 8 & !

# ) ,// / BD ( 7$908,>*0 D/ (

29,)*0F$ *9 .9 E ,& ) 7- 8

/ BD ( / ( 7$908,>*0 & ! ) D

%- -/ ^%-& ! -/ & !

% >% (

. 0D/ " & DD/ & ! *%! % "

. 2 .& ,&D&- &

*%! &

DAS A . " -0 B ^%- 7 C/R

<$1F()(> # " . 8 71 C%-8& !

) # " *%!

.

) & ! 3 *%*=

*%! 7 &

C%0 2 .& ,&--8& ! >!>%) #

*%! 7 & C%0 2 .& ,&-/8&

# 7 8

/ ,// 0// A// B// -/// -,// -0//

/ ,//

0//

A//

B//

-///

-,//

-0//

/ D/

-//

-D/

,//

,D/

C//

CD/

/ ,D D/

ED -//

-,D -D/

-ED

/ ,/

0/

A/

B/

-//

-,/

*%*=7%- 8

/ -/

,/

C/

0/

.,B/78 .0D/78 .DD/78 7$908,>*07S8

(51)

#N)#?<(#$1# #? # $<>># 0,

&&$* 0 6 :#

) & ; < 6 :# /

; <

) 7[ D/ W . 8 *%! . "

-0 B ^%- -- D ^%-

) #7 8 E DS% 1

) # 7 8 E DS% >

-,S% 1 & ! ) - , 7 8 - C 7 8

72 .& ,&-/8& ! ) C 0 7 8

72 .& ,&--8&

+ - 0 *%! U , C

*%! &

+ - *%! U ,

> . -0 40 ! 7) 8U C *%

! &

*%! "

6 7 = '8 4/ ! 7 = (8 C/

! 7 = >8 -E ! & *%!

(52)

#N)#?<(#$1# #? # $<>># 0C

( BB ! = ' C, ! = ( -B ! = >

&

H & * " !

! / : / : / ( 2) ! & !

! ( K )

;/ < ;/ < ;G< ;G / ^"< ;L <

. ) + ' "" 7"3 5 1 " "

( / !

8 " " 1313 7 * " 3

!

/ ( " 1 *5 7 " 3 " *

4 ( &

! / ( "* 99 "13 "* 9 19 1 3

G/( ))

) 2 "3 3 37 *93 " 7 13 * 1 9

K" G M " N/ ' / ^"& *

*$* A 0 0 0 & )

< ( " % .. *%! "

" ( " 7'P >( *'O8 7

& ,//C8& < ' ( " $%$% $%*%

% % " ; & @

*%! " #

> $C%

,9, $*,% $*C% 9 7 2 8 *%

*= . 7 & C%D8&

> " ,9, 9 *%*= & #

" ,9, @ 72 .& -&C&C8

7# ,//C8& ! ,9, ; > ?

*%! & < " " " >

" $*,% $*C% 9 *%! 7 & C%

D8& ! # " 9 *%!

(53)

#N)#?<(#$1# #? # $<>># 00

" < "

# " 72 .& -&A8&

9 " % # " "& & $

@ %> . = % ! 7 @ %>*!8 "

" # " %< 7' &

-44B > -4448& *%!

? =" 7<8 @ 7! & ,//,8&

" G *%

! & $*,% $*C% " G

*%! 7 & C%D8&

&&$* 1 A ! ! & )

! = *%! 7- ^%- D 29,)*0F$ *9%) D/ ( .9

E ,8 7 8 7

8 G =

? . & ! + 9 % . 7 8U ,9,%

. 7 8U-/ ( $C% 7 8U-/ ( $*,%7 8U-/ ( $*C% 7 8& <

6 9 9 %>. " )

*%! % . & ! "

. " *%! % 7-//S X -0 B ^%-

7 8 -- , ^%- 7 8 <$1%1

72 .& ,&D&-8 &

@ 7 8

/ 0/ B/ -,/ -A/ ,//

*%!%7S8

/ ,/

0/

A/

B/

-//

(54)

#N)#?<(#$1# #? # $<>># 0D

< > 9 *%

! % &

*$ 2 &

< ? "& & ! *%

!

' " " $, " " & ! "

" %? 1 C%, " &

H & * # ) 2 @ /( ) // /&

2

2 . "

*%! ! $ $C $C%%? ⁸

T ⁸ T T CD ,C

T⁸ T T CE AD

T T % & &

T % T & &

% T T & &

% % T & &

< " [ - 29,)*0F$ *9%) 7D/ ( .9 E ,8 - *%

! -/ ( $ $_C D ( ! # %!

" %? 72 .& ,&D&,8& ! " %

? . 2 .& ,&A&, &

8 $90T %- %- A/R

8 *%!

T

%

& &+

(55)

#N)#?<(#$1# #? # $<>># 0A

&&$* 3 '

; < ; <

= 2 +

! " [ - 29,)*0F$ *9%) 7D/ ( .9 A D E ,8 -

) -/ ( $C% D ( ! 72 .& ,&D&,8& ! " *%

! % H-//S X -0 B ^%- 7 8 -- D

%- 7 8I " %? *

<$1F)(>%1 72 .& ,&D&-8 72 .& ,&A&,8&

> + 7 8 $90T% .9 E ,U 7 8 $90T% .9 A DU 7 8 *%

*= .9 E ,U7 8 *%*= .9 A DU7 8 $90T%

$_C^%72 8 7 8 $9₀^T% # " 72 8&

/ / / D - / - D , /

/ ,/

0/

A/

B/

-//

/ -/ ,/ C/ 0/ D/ A/

/ / / D - / - D , /

/ ,/

0/

A/

B/

-//

@ 7 8

$90T %7(?"%- 8 $90T %7(?"%- 8 *%*=7S8*%*=7S8

(56)

#N)#?<(#$1# #? # $<>># 0E

!

"

< " *= *%!

& @ " <$1%1 7

( *%! % 8 *%*=

& "

@ " %? " & " *%

*= & & C%A " *%*=

" %? & ! ! % " *%!

"

*%*= &

" " & K . " %?

9 *%*= & #

(; # *%*=

? " " & "

" K . * =

*%! &

*$ $" 6 2 +

! > ; " " 7 > 8 "

7 ) 8 ,+-

9 " & ! ; ) ;

! & @ K . G %( "

72 .& ,&-C&-8

7 & C%E8& ! "

2 . *%! ! " &

! ) . % "

" " &

# " -D ( " D ( " & < "

*%! ! " &

(57)

#N)#?<(#$1# #? # $<>># 0B

&&$* 5 : & = 2 + !

G " %? "

G % 6 70/R 8 72 .& ,&-C&-8& ! . ? "

, 29_,)*₀F$ *9%) 7D/ ( .9 E ,8 , *%! -/ (

! D ( 7 8 -D ( " 7 8& 2 + # " 7 8 "

7 8 ! 7 8 " ! 7 8 # " ! 7 8

# " " 7 8 ) 7 8&

$ CD < " #=. & ! " -D

( " CCE 4D ± , AC W 7CD4 DB CCA C, W 8

. -C ,0 ± / DD W 7-0 /4 -C -B W 8 " & -D -A ± / - W

7-D /A -D ,B8 $90T& ! " D ( " -CA BD

± D -E W 7-C- AB -0, /, W 8 U . D CA 7± / ,/8 W

" & D CA ± / , W 7D -A D DA 8 $90T& #

" %? 6 & < "

*%! " !

% & #

" $90T% " "

? ? & !

. ! &

! > ; > )

&

@ 7 8

/ D -/ -D ,/ ,D C/ CD

%7W8

/ D/

-//

-D/

,//

,D/

C//

CD/

(58)

#N)#?<(#$1# #? # $<>># 04

*$ $ >! / 2 +

> ; ? "

) " G 7B 8

& ! 0 29,)*0F$ *9%) 7D/ ( .9 A D8 -

*%! ^%- D ( ! 0 E D W " & "

# 9 =

& ! G D/R 7

8 " & A/R 7 8 & <

6 " G

& ( 9 9 %>. " D/ W

) & ! " %

% 72 .& ,&A&- 2 .& ,&A&,8 &

. 72 .&,&-C&,8&

< & C%B *%!

E D W " " E AA 7± / /C8 W $90T E E4 7± / -C8 W $, "

71 C%C8& ! *%!

? 0 W " " C B/ 7± / /E8 W $90T C BB 7± / -/8

W $, 71 C%C8& ! *%!

" %? 7 & C%B8& #

" %? ) $90T

$, -+- &

- $

C%

T 0 9

^T

T ,

^%

/ 44 $9

0T

T - /- $

,

H & * * >! / 2 +

*%! "

7W 8 7W 8 >

7W 8

< << < <<

E D E AC E AB E AA E 4-

0 / C BE C EC C 4B C E4

(59)

#N)#?<(#$1# #? # $<>># D/

&&$* 7 >! / ! 2 + !

; <

! " [ 0 29,)*0F$ *9%) 7D/ ( .9 A D8 - # "

%- D ( ! E D W " 7 8 0 W " 7 8& "

= 72 .& ,&D&,8& ! ) G

D/R 7 8 " & A/R 7 8 & !

$, . J1 ! L

3 " 72 .& ,&-C&,8& !

" %2 " 2 .& ,&A&- 2 .& ,&A&,

&

> + 7 8 " %? 7 8 $90T% U2 + 7 8 " %?

# " 7 8 $90T% # " 7 8 $,% # " &

@ 7 8

/ - , C 0

/ - , C 0 / , 0 A B

"$9 0T $ ,7W8

7 8

"$9 0T $ ,7W8

(60)

#N)#?<(#$1# #? # $<>># D-

*$ $* @ 2 +

! . " %? .9%G A D

1 . A/R & ( > 7-4B/8

1 . . *%! % ACR &

&&$* 9 & // /& !

; <

; < 2 2

! " [ - 29,)*0F$ *9%) 7D/ ( .9 A&D8 D (

! - *%! , 0/ ( " 7 8 2 7 # " 8

7 8 72 .& ,&D&,8& ! ) A/R &! "

72 .& ,&A&,8& ! < " 9 %) & ! !

( " #=. &

N₃^- (mM)

0 10 20 30 40

$90T%7%-%-8

/ -/

,/

C/

0/

D/

A/

;<?

" *

3 7

0_* ;/%<

/ -/ ,/ C/ 0/

$90T%7%-%-8

/ -/

,/

C/

0/

D/

A/

0* ;/%<

" *

;<?

3 7

0* ;/%<

7 8

(61)

#N)#?<(#$1# #? # $<>># D,

! %G " %? .

9 7-4C,8 & # %G A /A ( 7 & C%48

0 4C ( &

! %G " *%!

. .

" " & ! =

" %? # " D0

$90T %- %-& *%*= 7-, ,C W

%- %-8 " %? C// &

*$ $ 4 # )B 2 +

*%! * 9, # %!

! " & G ! # %

! " %? &

# %! * 9, 1 7<<<8% " %?

&

&&$* " 2 + / ! ! #

!

! " [ - 29,)*0F$ *9%) 7D/ ( .9 A D8 - 0 # "

%- -/ ( " # %! + D ( ! 7 8U9_, 7 8U

* 7 8 - ( 1 7<<<8% 7 8& " =

72 .& ,&D&,8 " & *% . * # %!

9,% . 9, # %! & ! ) A/R & !

2 .& ,&A&, &

@ 7 8

/ -/ ,/ C/ 0/ D/ A/

$90T %71"%- 8 / D//

-///

-D//

,///

,D//

C///

CD//

(62)

#N)#?<(#$1# #? # $<>># DC

& C%-/ " # %! "

" %? & 9, # %

! " %? & ! " %? 9,

# %! & B/S ! &

! *%! 1 7<<<8% # %!

" %? & # *

? 7 & -D 8 # %! 7 & C%-/8&

) " *%*=

*%! < " * &

&&$* "" ' = 2 + !

= 4

! ? " [ - 29_,)*₀F$ *9%) 7D/ ( .9 A D8

- # " = & ! )

9, * 7, 8& ! = "

7# " -/ (8 9 9 %>. " " & ! )

G 7D/R 8 & ! " . "

3 * → <$1F()(>%1 72 .& ,&D&-8& H-//S X

,- D ^%-7 8 -, D ^%-7 8I&

+ 7 8 9_,% . 7 8 *% . U +

7 8 9,% . 7 8 *% . &

@ 7 8

/ -/ ,/ C/ 0/ D/ A/ E/

*%!%7S8

/ ,/

0/

A/

B/

-//

(63)

#N)#?<(#$1# #? # $<>># D0

< " " " 9,% . "

*% . *%*= *%!

& " *= 7 & C%--8

? & ! 9

" *%*= " %? @ &

# *%*= *%! *

. *, 9, . *, 7 ,/// ' ,//,8&

< *%! " F * "

# & # ; ;

% 6 >. *,

" * < &

" " 9,% . 9

*, 9, " *%! 7'

,//,8& !

" * *%! &

&&$* "

! ( 2 =

! ? " - # " - 29,)*0F$ *9%) 7D/ ( .9

A D8 D ( ! -/ ( " & ! ) =

* & ( *%*=

? # * ..& ! "

. " 7-//S X -0 D ^%-8 3 * →

<$1F()(>%1 72 .& ,&D&-8& > + 7 8 " * " U7 8

" " U7 8 " " ! &

@ 7 8

/ ,/ 0/ A/ B/ -// -,/ -0/

*%!%7S8

/ ,/

0/

A/

B/

-//

(64)

#N)#?<(#$1# #? # $<>># DD

# < @ !

7 & C%-,8& " *

K . " %?

9 & <

) 72 .& C&0&A8&

*$ $1 + 2 / . /

! *%*= %

7 ,/// ! ,//, # ,//C8& ! <

@ 1

2 . ( " 72 .& -&C ! ,//,8& !

" @ *%!

7! ,//,8& <

*%*= " %? 6 2

" *%*= " %? &

# 2

" *= % ? *%!

# " > ; &

*$ $1$" 2 + &

! " ( " *%! %

( " "& & N %*=

N %! 7 ,/// ( > -4BC8& !

*%! > " &

5 > %'

? 7> $^%8 % $

@ *%! 7! & ,//,8& !

# " *%*= & #

" " %? *%

*= @ 72 .& C&0&08& * " %

? @

(65)

#N)#?<(#$1# #? # $<>># DA

*%*= . 7! & ,//,8

# > %' % @

&

! *%!

-/ ( 2 72 $8 & G

@ ) " ( *%*= % " %

? " > 72 .& ,&-,&-8 &

! < *%! 2 $ =

" < &

! 9 " *%*= " %

? & C/ 7 & C%-C8& ! ( *%

*= % " %? #

2 $ > . = %,D 7)!%-/8&

! *%*= .

" %? & ! " ? 7> $^%8

. . " 7 & C%-C8& ! ?

@ % # " (

C/ ! " & $ ? 7$*8C

? 7> $8,

. = 0A/ " & # . F %

>. C// A// %

*%! &

$ < *%!

7 + -0 B ^%-8 2 $ @ -4 ,0

> 7- EB ^%- # " 8 # "

7 & C%-C 8& 6 % 7- D0

%- # " 8 ) & ! "

. 9 *%*= % " %? & *%

!

" ? 7 & C%-C 8& # "

@ *%! *%*=

" %? &

(66)

#N)#?<(#$1# #? # $<>># DE

&&$* "* . ! ) '

2 +

; < ; <

8 2 ! & ! = )

*%! 7, , ^]-8 = -/ (

" 72 .& ,&-,&-8& ! " . " *%! %

72 .& ,&D&-8 " %? 3

? & ! " " %?

[ - 29,)*0F$ *9%) 7D/ ( .9 A D8 -&/ # " -/ ( $C%

D ( ! 72 .& ,&D&,8& ! " %?

2 . ,&A&, & ) "

? " 7 " C/ ! 8 ?

3 " 72 .& ,&-,&-8& H-//S *!9 " & $90T%

X -0 B ^%- ,B , ^%- ^%- 7 8 -- D ^%-

CD B ^%- ^%-7 8I&

> + 7 8 *%! % U7 8 U

7 8 > &

0 4 8 12 16 20 24

/ ,/

0/

A/

B/

-//

/ / / 0 / B - , - A

Time (h)

/ 0 B -, -A ,/ ,0

/ ,/

0/

A/

B/

-//

/ / / 0 / B - , - A

@ 7 8

*%!% $90T%7S8*%!% $90T %7S8 > 7%- #" 8> 7%-#" 8

7 8

(67)

#N)#?<(#$1# #? # $<>># DB

*$ $1$ 2 + ! &

% *%! " *%*= % " %

? & % > %

' @ " " %?

*%*= & @ 2

@ *%!

H >( *,I% 6 9 & < - ( %

" @ = *%

! *%*= 7! ,//,8& !

9 % @

*%! 72 .& -&C&,8& ! "

@ # " %

7! & ,//,8& & C%-0 " " %? %

*%!

&

= *%! -D

? . 2 " % 7/

- , (8 & # *%*=

" 2 " % 7 & C%

-08& *%! / /D ( %

" & D/S *%*=

0DS " %? 7 & C%-0 8& ( % %

2 " 7/ A (8 ?

- BS " & 0 ,S & ; 2 " %

& *%!

% "

? 7 & C%-0 8&

# % *%!

@ *= %

? & ! @ *%!

*%*= " %?

" @