免费文献传递   相关文献

Cellulase Produced by the Symbiotic Fungus Leptographium qinlingensis of Dendroctonus armandi

华山松大小蠹共生真菌秦岭细粘束孢产纤维素酶的研究*


通过单因素和正交试验对秦岭细粘束孢分泌纤维素酶的组分、条件等进行研究。结果表明: 秦岭细粘束孢可以分泌内切葡萄糖苷酶(CX)、外切葡萄糖苷酶(C1)和β-葡萄糖苷酶(BG),具有完整的纤维素酶系; 其中CX是秦岭细粘束孢分泌的主要纤维素酶组分,优化后CX酶的最大酶活为35.215 U,分别是C1和BG最大酶活的13.662倍和8.780倍。正交试验表明:秦岭细粘束孢分泌CX,C1和BG的最适条件各不相同,产CX的最佳条件为29 ℃、6天、起始pH4.0、摇床转速150 r·min-1。温度、转速、起始pH和时间4个因素对秦岭细粘束孢产3种酶的影响各不相同,影响最大的是温度,其次是转速和时间,而起始pH对该菌产酶的影响最小。

Components of cellulase produced by Leptographium qinlingensis and the optimum conditions for biosyntheses of the enzymes were studied by single factor and orthogonal experiment in this paper. The result showed that Leptographium qinlingensis had a complete cellulase system which was composed of endo-β-1, 4-glucanases (CX), exo-β-1, 4-glucanases (C1) and β-1, 4-glucosidase (BG). Furthermore, it was also showed that the enzyme CX was the major component of the cellulase whose optimum activity (35.215 U) was 13.662 times higher than enzyme C1 and 8.780 higher times than enzyme BG respectively. A orthogonal experiment indicated that the optimum conditions of enzyme CX, enzyme C1 and enzyme BG were different. The optimum conditions to produce enzyme CX were at 29 ℃ with six-day cultivating time, pH value 4.0 and 150 r·min-1 rotating speed. Among the four factors (temperature, cultivating time, pH value and rotating speed), temperature, had the largest impact on producing cellulase by Leptographium qinlingensis,followed by cultivation time and rotating speed.


全 文 :! !"" ! #$#
% $ $ &$ #$ %
& ’ ( )
’()*+,)- ’)./-* ’)+)(-*
/012!"*+02#$
3456*
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
% $ $ &
>(P*09vXlc%0*:+;=
wOA"~QR"
/Rh# ? E% S T% T E# ,UJ# ?1O#
F#2 xd_&(t*)X=())H ‘a 7#%#$$T%2 xd_&(t*)&)H ‘a 7#%#$$f
! "# 0T6zAy^(<_m%0*:+;@;LY@X%0*:+;@<~NOwOA"D@*<;[ (8 "~)*".g <"2%#" =*@/X (# y :;)*".~
#<2>>%,y ?27?$,P^(<_STU%0*:+;@< (8*(# y :;~)g5AMYZh*= (8 ~)I5Ag
%& @x>3xIW AB!2$x|-|? #"$ CHDEF9 #Pw,x|?xIW AByiR !-zAm%0*:+;= <1"~K
LMYZh*KL)*~Xw,*BfX|?yiR*GIW ABmoc="~KL)0P
$%&U >(P*09T%0*:+;TwOA"T^ (<_
’()*+#’7><2#" ,-./0#- ,12+##$$# 9 7!??3%$$&4#$ 9 $#$? 9 $"
|&’#U%$$? 9 $& 9 $!P
j()*UGU+4()j(\])*F<$7<$$7"YGg01?=P
!"##$#%&" ’()*$+"* ,- ./" 0-1,2).2+ 3$45$& !"#$%&’(#)*+, -*./*.&".0*0
)6 1".2’%3$%.+0 (’,(.2*
;G0 HIEJEG# (KLF BIE% ,GFM NEFM% 8I BIE# .E ’K0IAEFM# (KLF ;IEDLE#
F#6 !"##$%$ "& ’(&$ )*($+*$,*-"./01$,/ 234 5+(6$.,(/7 89+%#(+% 7#%#$$T% 6 !"##$%$ "& 4".$,/.7*-"./01$,/ 234 5+(6$.,(/7 89+%#(+% 7#%#$$f
7,&.(%+.U (0DA0FLF5O 0P 4L11I1GOL AC0QI4LQ RS ’$:/"%.9:0(;< =(+#(+%$+,(, GFQ 5KL 0A5EDID 40FQE5E0FO P0C RE0OSF5KLOLO 0P 5KL
LFTSDLO ULCL O5IQELQ RS OEFM1L PG450C GFQ 0C5K0M0FG1 LJALCEDLF5 EF 5KEO AGALC 6 ,KL CLOI15 OK0ULQ 5KG5 ’$:/"%.9:0(;< =(+#(+%$+,(,
KGQ G 40DA1L5L 4L11I1GOL OSO5LD UKE4K UGO 40DA0OLQ 0P LFQ0V!V#*!VM1I4GFGOLOF(8f*LJ0V!V#*!VM1I4GFGOLOF(#fGFQ!V#*
!VM1I40OEQGOLF:;f6 WIC5KLCD0CL*E5 UGO G1O0 OK0ULQ 5KG5 5KL LFTSDL (8 UGO 5KL DGX0C 40DA0FLF5 0P 5KL 4L11I1GOL UK0OL
0A5EDID G45EYE5SF<"2%#" =fUGO #<2>>% 5EDLO KEMKLC 5KGF LFTSDL (# GFQ ?27?$ KEMKLC 5EDLO 5KGF LFTSDL :; CLOAL45EYL1S 6 -
0C5K0M0FG1 LJALCEDLF5 EFQE4G5LQ 5KG5 5KL 0A5EDID 40FQE5E0FO 0P LFTSDL (8*LFTSDL (# GFQ LFTSDL :; ULCL QEPPLCLF5 6 ,KL
0A5EDID 40FQE5E0FO 50 AC0QI4L LFTSDL (8 ULCL G5 %& @ UE5K OEJVQGS 4I15EYG5EFM 5EDL*AB YG1IL !2$ GFQ #"$ CHDEF9 # C05G5EFM
OALLQ6 -D0FM 5KL P0IC PG450COF 5LDALCG5ICL*4I15EYG5EFM 5EDL*AB YG1IL GFQ C05G5EFM OALLQf*5LDALCG5ICL*KGQ 5KL 1GCMLO5
EDAG45 0F AC0QI4EFM 4L11I1GOL RS ’$:/"%.9:0(;< =(+#(+%$+,(,*P0110ULQ RS 4I15EYG5E0F 5EDL GFQ C05G5EFM OALLQ6
8"- 9)(*&U >$+?."*/"+;, 9.<9+?(T’$:/"%.9:0(;< =(+#(+%$+,(,T4L11I1GOLT0C5K0M0FG1 LJALCEDLF5
%0*:+;F ’$:/"%.9:0(;< =(+#(+%$+,(,fX>
(P*09F>$+?."*/"+;, 9.<9+?(fmtVcho2
~21vXlc*D>(P*09un,’cN>
(PF@(+;, 9.<9+?(fi/1[U8=fyy8Sy
YyD2R*vhrpcNU8~hi@i>$c
NU8~;(PwOAy8
SA0;~qCK*L@8SA"y-H#-"FYG(*%$$!T(KLF $/ 9# 6*
%$$7f*m>(P*09D>(P"~tia\O~
?$P
*y@:uHPt#}>x09txU!(VX
Y:;FGgu*5FCm45;S0YK;>$*
QNBvXlcm:;NOm@8SAywOA~
{iG>$FZ0RGSGOKE $/ 9# 6*%$$?fPEi?(
F%$$!fQRh@tAxB<8pyr- < 1K1~
!DH1}m}>qRwOA".H~KL*\v
$K;bLwOA";GWX%L}>~rpT
[LFM (F%$$?fVch$Z.}>F A?"+/"/$.<$,
&".<",9+;,fABvXlc B$.<(/"<7*$, 9#C;<(+",;, ~
wOA".HPC-b(F%$$!f@;a;h/)D
34F !"#$%&’ ()#*’#$f-t~RB89%F"9Q
RhBHSTv*B(F!""#f#N*<=h(Z;
{mPx34F+%&%,-’*./ ’01)#&’1./fwOA"~K
L*5Xl[09tvXlc~wOA"QR;U
H<=P9sD3;@2qIJ%0*:+;wOA"SRD>(P*0
9unrp>(P7~?$P
$ yXavv
!"! !}4V)qr}
O)H!X;QR‘]OPOjF%&’("fF)*+,-.*!""’f*$[c1~.;N@5I~tAEN!"# 456A78
0%0*:+;/1[tAEN< /3[@u!c|uK;{*yu&bjk9~
UQ>7*‘a*$!c|jm $"# -H123 $~;{
t F^4., )1 ’0 5*!""6fP
!"$ 9Y6A78)[(8AHj
D !’" 12 UQ>7& $"" 12 OF="N<
j*/u;{t^ 7 12*!/ 0x$$" +H1893 $*‘aN
< #3POF^[: 0*’ """ +H1893 $;t $" 189*
":^qgO"^PcGqp[>f"lQ3>*
$!cB;{|!u*6" 0WT~S}[8B+
S}P
!"% \YDEA+v
0";LEO"^~w,ig !"*!7*!’*!/*!;
y7$ 0F%&’("*$$" +H1893 $*#3f*iRg !*:*#*
6*$" y $! 3F%&’("*!/ 0*$$" +H1893 $f*%& g
7(’*:("*:(’*’("*’(’*#("*#(’ y /("F!/ 0*$$"
+H1893 $* # 3f*| ? g ;"*$$"*$7"*$’" y
$6" +H1893 $F!/ 0*%&:(’*#3f*@/i8Yhw
,xYhiRxYh %&yYh|?m="~KLP
D";6zA<_~j!"V :zAFw,xiRx
%&7x|?f7|b~^(<_P
!"& :;zYZPHj
Zu~wOA"X)?$v!Yh5=Z/H
h_;|iwOA~ 7^"Dm*qRB89%F
"FG8 <=*><7(!($(:fT?B89%F"FG8
<$*><7(!($(;$fy! 3 89%F"FG8 ?4*
><7(!($(!$fF@,,- )1 ’0 5*$;/6fP!wOA"
FAB.CDfY ’" 1Ew>>fg0;T<= $ ! FF2 G3f
OojwOA F^!QwOAg0;T?4 $ !FF2 G3f|‘Fg0
;*y$ "("’ 1,KH23 $x%&:(6~pH- 3pH-^
$!^jLPn:qK@ $(’ 120;y "(’ 12O
"^*’" 0bwFAB.CD" #" 189*BC" 7" 189f*M
?$"|8TT‘wT$@u 7 12 LJM I|&
’ 189*#+[,BT !" 12*[ /’!J32?LH@
33,q"8t NL’:" 917F4O,CD*$;6/fPYrJz
"^r@==X $!E89%FYjOF^7~;V
%ftGg2-".6(FPfPY"<_y\&
7fP
!"’ -./0
#$ >QRDKFS!""7f@AopLq@B9
QmP
! QRa@B
#"! qr=b5\YA;<
45N+[{f*<= D!/ 0i".WX)+F!(;$7 Pf*<$
y AB.CD~".D!; 0y!7 0iWX)+Pf[
!’ 0ii8YX<$ ".H*4L=Xw,cfKL
cGq~m9X0*ES=".cfP5X ?4 "
.H#45w,~z+G{f*!; 0ii8YX
?4".HP7$ 0i : 1".HyBi8YX
Fm $.f*5T%0*:+;Dfw5AK="LR
caPM1w,5AK <= ~".Hy+[BC"*
X%0*:+;@<~NOwOA"1^*gb9
<_t‘ <= "gNOEom-op<;P
#"# qrfR5\YA;<
N~".cf*42i#:X%0*:+;~g:#
y>?X0SWPN<3#F: T # 3f4iR2@
c}342@*bSWcqX0m9*="}+D2
**".4Wz+PN< # 3i*<= ".yX;}
yWX)+*".g 6(/6’ P*X;}g #("66
EH23 $P#3[="=sM?K{xX0mPPT! 6
3X;}gb"z*G 63[#ONK{Tbi*4
5iR~2@*cqX;}y".yjK{~=P
N< 63i AB.CD*?4*<$ ~".WX)+*V)*
X;}bSN[T6 T $" 3~".Zmc,t
Fm $UfP
#"$ qr}@< ()5\YA;<
hN".j"z~=Th %&g ’("i <$*?4y AB.CD"
.WX)+*@/g 7(’#"*"(/:"y "(/:" PTG <=
;"$! $"# +T’(U>(P*09vXlc%0*:+;=wOA"~QR
)*".7lkD !"#$%*g &%$%’’ (TD !"#$% )
%$%~9:R *+ j9bM,tT!"PT %$%i*"
.EA{fT5T%-HF!"#$% ) %$%f5AgEo
c="PIW !"f[ #$,i*="=s<*/"p
L.TD !"%$% ) -$%9:R*45 !"2**".E
AK{Fm ./fP
m ’ N1234’ 566789 :6 8;<92=/92:> 97?!7@/9;@7 />A 92?7 :> !@:A;82>3 87<<;$!$1E/B7 5>FD?7 1EG*$"$*’ " 5>FD?7 *’*$#$HI" 5>FD?7 HI*$"$*+ " 5>FD?7 J+*$K$X;} LD87<2/ C2:?/BB4Kh MN7 B/?7 C7<:O4
m . IW !"x|-|?m*:+;=wOA"~KL
1234. 566789 :6 2>292/< !" />A @:9/92>3 B!77A :> !@:A;82>3 87<<;!"# =>|p5\YA;<
h|-|?g P, @H?2>0 ’ii8YX".HT
h|-|?g ’%, @H?2>0 ’i= HI*1E/B7LR)I*
f[W+[b|?*="|bVoK{*5X= *+*
*’ )ILR~|?g ’’, @H?2>0 ’P*+ ".D ’’,
) ’&, @H?2>0 ’WRj9,t*GD ’%, @H?2>0 ’iB
".Jg ’’, @H?2>0 ’i~ #P QTD ’R, @H?2>0 ’i
#Bi8YX *+ ".*biS8~ *’*HI".+
@/(b)+".~ #’Q*#-QP|?)+oCD
skVBsT*GkmcGN^*tBX0;Xb
LoGKLX="|b*gb9QRtu ’&,
@H?2>0 ’g)I|?Fm .CfP
!"$ ?@}~
;S ’LYll*%0*:+;LY@89%F"F*+fx?B89%F"F*’fy!089
%F"FHIf*abZu~wOA"KTB7 *+ ".
Zmc+*XNO~wOA"D@*<;[ *+ ~)
*".@/X *’ y HI )*".~ ’&$--. ,y
R$SR,,P^(<_*l%0*:+;@< *+**’
y HI~)g5AMYZhP= *+ )I5AgU
.P Tx-3xIW !"#$,x|-|? ’%, @H?2>0 ’T=
*’ )I5Ag.P Tx%3xIW !"%$,x|-|? ’&,
@H?2>0 ’T= HI)Iw,g.% T*BC5Ah *’P
b?*#-zAmM".~KLMYZh*j]r U
7~*0Wc4;#KU’f1E/B7y *’Uw, V |
? VIW !" ViRT.fHIUw, ViR V|? V
IW !"T&f*+U|? Vw, ViR VIW !"P!
~gl*moc="KL)*~zAXw,*BfX
|?yiR*IW !"moc~="KL)0P
& Qca]c
w,m%0*:+;=wOA"~KL)**
w,~:;ath%0*:+;@^xZm".~*0A &1".~VLT4+R/
,’’ & ’ ( ) #%"
C ! ?@}~efAAP)T
"#$% ! &’()*+* ,#-.* #/#-0121 34 3)(5363/#- *’7*)2+*/(
<_;I
!"#$%&’$()
*+,
<_zA !"#$%&’$() -./)+% ". !(01’$ ./)&2&)&$34F5f
w,F6f
7$’#$%.)8%$49
iRF:f
7&’$4; #<
F=f
|?F>f
?+).)&(@ 3#$$;4F%H’&(A Bf
CD.3$ =B :E =F
B BFGHf BFHf BFIJKf BFBBKf GJKLH BJBBK MJNHH GOJPNH
G BFGHf GFOf GFIJHf GFBMKf BJHNP BJNKM MJKOL BNJKKK
M BFGHf MFPf MFHJKf MFBHKf GJBLI BJBBK GJHNL GIJGNN
I GFGPf BFHf GFIJHf MFBHKf KJOGG KJHHH GJBBK GPJHKO
H GFGPf GFOf MFHJKf BFBBKf KJOOG KJLPB BJIBO GPJOII
O GFGPf MFPf BFIJKf GFBMKf KJOOG KJOLI BJGKN BHJONP
P MFGLf BFHf MFHJKf GFBMKf KJNKK GJHPN IJKBB GMJHKG
N MFGLf GFOf BFIJKf MFBHKf GJMNB KJHHH BJLPB MHJGBH
L MFGLf MFPf GFIJHf BFBBKf KJLNO GJGGK GJKNB GNJPKN
QCD.3$B HJNPH MJHBN HJBMN MJPIM CD.3$.H 6/)&2&)1 +- CD6
QCD.3$G BJLIO IJOMK MJBLI MJKIL NfzA R.&( -./)+%Uw, S|? S #< SiR
QCD.3$M IJBOP MJNIB MJOHO HJBLP 7$’#$%.)8%$ S ?+).)&(@ 3#$$; S #< S =8T)&2.)&(@ )&’$
? MJLGL BJBBG BJLII GJBIN )<|b U#)&’8’ /+(;&)&+(3U6B:G=B>M
Q=BB IJKGM IJGIG GJMHN IJMKK =B ".H 6/)&2&)1 +- $(01’$ =B
Q=BG GJGBL MJMGL IJHPN HJKPI NfzA R.&( -./)+%Uw, S|? S #< SiR
Q=BM HJMHG IJKGM IJOHN GJGBL 7$’#$%.)8%$ S ?+).)&(@ 3#$$; S #< S =8T)&2.)&(@ )&’$
? MJBMM KJLBM GJMKK GJNHH )<|b U#)&’8’ /+(;&)&+(3U6M:B=M>G
Q:EB LJHBM LJLPO PJKMI PJMHG :E".H 6/)&2&)1 +- $(01’$ :E
Q:EG IJPMI OJIHO PJGHL NJGNN NfzA R.&( -./)+%Uw, SiR S|? S #<
Q:EM NJKOM HJNPN NJKBP OJOPK 7$’#$%.)8%$ S =8T)&2.)&(@ )&’$ S ?+).)&(@ 3#$$; S #<
? IJPPL IJKLN KJLNM BJOBN )<|b U#)&’8’ /+(;&)&+(3U6B:B=M>G
Q="B OLJKPG PPJPLG PPJONP NMJBMP =F ".H 6/)&2&)1 +- $(01’$ =F
Q="G PKJNMP NKJNHL PIJGBM HPJBNN NfzA R.&( -./)+%U|? Sw, SiR S #<
Q="M NPJIGH ONJONM PHJIMI NPJKKN ?+).)&(@ 3#$$; S 7$’#$%.)8%$ S =8T)&2.)&(@ )&’$ S #<
? BNJMHM BGJBPO MJIPI GLJNGK )<|b U#)&’8’ /+(;&)&+(3U6M:G=B>M
5TYhw,5AK*%0*:+;~SRmcN
U8~KLYh*m>(P*09uni~H-?
$SbZ::;*zba>(P*09un~iQ
@Eb2tlKP<&/V$(FGKKOfq.lZ[w,
~:;KL$(*0 F!"#$%&’(&#)* +&#$"%&*,"f~
eOPW&"(FGKKPf+Ok$(*09yF~ G 1
vXlc -%&*.,##/, ’0,1/2"%,*3+4/&*(&., .&#(/). W
R~lK4w,Yhbq*:;P<+-3)$))$% (
FGKKPf~QRSST09tmU8~np4iR~
:;G:;*BvXlco4w,xU8B2~;)
;SxABC!X;~YhGbS:;P2G
FBLLPf0TQROkKLBM09F 5&./’)*
+/#/+"%$,f~NOSXc 6"+(&2%,+4/). 7/#28/"0$// D
Yhw,KmKt|PF9/#)* *:01"*(%/*f~n&sy
DAsbr/P
9p~OXX09tmU8~rpxcNU8
~[:n:YAvXc~n&U=Z/?$~Q
R*zbbL[cO;txcxUU=gQRP
X.’.’+)+(FBLNOf.lwOA"~?$oCDPU
pNTQ8%+;.FGKKHfOk%:c ;"%,(&’:*(/* +&0&#/’,
~fXSR=;oKL’92’F9/’", <"=&"#*/*f~
|@":*)FtBL|pNTR!Y(FGKKPf+

$cNrs2’8Sy~wOA*X%&O&([
Rrs2’F9/’", ,>/"*fDH~\OAVcP4V
5Tlc~SR=;[BXwOA"mcNU8~
*Hbq?~E^s*m09trpcNU8IO?$PY& (FGKKHf]l0Tm1[>F ?&0"#&+*/*
/#1/’(,fI=vX*c~QRgK_w~1[>X;
[cvvPhu+LY;lc~Q,g\v0TK
;bLvXc=XwOA"*;GWXm09t+
bL~X;[cPb?*o8HptqRYAaB
vX~lc7jb~MNwOA"=*7*o2q
QR\]a>$P
] ^ , -
Y G*m ‘ , GKKK , >(P*092cy(AVchQMQR ,
&’()*MOFBfUHM A HP,
Y G*P T*|0=*( , GKKI , >(P*09yvXlc@<
"Dm@B , &’()*IKFHfUBGM A BGO,
v*B*/Q.*Y9m*( , GKKO , Px34wOA"~QR! ,
;q5AK(Z;{mPx34wOA".H~KL , &’(
)QR*BLFGfUGKH A GBK,
BBB! BK# +T’(U>(P*09vXlc%0*:+;=wOA"~QR
R!Y*d-.*!"#$ %*( & ’(() & %:lcHI~rs2’8S
yiD)@AAwOA".H~:; & &’()**+F,fU-* .
/(( &
2 G*01#231#4 5& /--) & SXcF! & "#$%&#’()##fmKt|PAp?$
QR & W&At01**U’+ . ’6&
C-b*/7’*$%A*( & ’((* & /)D34-tRB .!. /*
* .8)%"~a;AHS & &’()**(F’fU/(+ . /(, &
Ei?*&{$*‘3F*( & ’((* & U1Tt1!DH1}m6{
m}>>>qRwOA".H~KL & _K))<*,F’fU
)’ . )6&
78#9 :*;<9= >*012 0*’* +( & ’(() & 7?3@A8#B1A<$ $@A<$1C<31@9 @D F8@GF8<3!’1*2%/+13#-0 4#$(#$%’$.#. & %?BI1@G1G**+U,6 . )(&
J#9= ; 5*78#9 7 K*78#9= > 0*’* +( & ’((L & J1DD#4#9A#G 19 A#$$2$;#4B131E<#f <9E 38# G?BI1@31A D29=2G 6’/0#*2078’. +(9-0#$2.-. &
%@A1@I1@$@=?*6F+fU,-) . )(* &
N<@ O >*P#9= : Q* R82 J :* ’* +( & ’((L & S4@E2A31@9 <9E
A8<4:.1’/%#((-. *’//’-. >// 29E#4 G@$1ETG3<3# A2$31H<31@9 @D A@49 G3@H#4 &
Q1@4#G@24A# ;#A89@$@=?*--U),’+ . ),’- &
N8@G# U& /-L) & >#78#B1G34?*6-F’fU’6) . ’,L &
:1AV# O !*0@=<9 O !*S@W#$$ O X*’* +( & ’((, & 78<9=19= 3#BF#4<324#G
19D$2#9A# G21312$)’/2.+’f @23I4#N#@F8?G1A<$ K#G#<4A8TQ1@=#@GA1#9A#G*///U/ . /’&
:@DG3#33#4 K P*J#BFG#? ; J*Z$#FC1= Z J*’* +( & ’(() & ;#BF#4<324#T
E#F#9E#93 #DD#A3G @9 B232<$1G31A* <93<=@91G31A* <9E A@BB#9G<$1G31A
193#4*) . 6,&
Z@IB?A<9=1<$ D29=1 3@ $<4H<$ 9234131@9 @D < $#[A@$@=?*’’U)// . )’’ &
Z24@E< Z& ’((6 & ]?$#B E?GD29A31@9 19 \#C@ GF42A#F ,#8’+ <’=2’$.#.f19@A2$<31@9 W138 38# I$2#TG3<19 D29=2G >’/+*287.*#. 12(2$#8+ & [24 O 5@4
S<38*+6U+*, . +6L&
01 :*>#E19< 5*^ 19G@9 % Q*’* +( & ’((6 & MG@$<31@9*A8<4B@$#A2$<4 1E#931D1A<31@9 @D IF?2(’$21.#. #$@#8*+fB1E=23 & O@249<$ @D M9H#43#I4<3# S<38@$@=?*L-F+fU
’(+ . ’(-&
%1" J 0*Q#93C Q O& ’(() & ;#BF#4<324# E#3#4B19#G G?BI1@93 B2$31F<4313# I<4V I##3$#TD29=2G #A3@G?BI1@G1G& >1A4@I1<$ [A@$@=?*6*
F/fU//’ . //L&
P@@E ; >*>AA4<# % M & /-)L & ;8# A#$$2$F241D1A<31@9 <9E F4@F#431#G @D G@B# #9E@=$2A<9GF#A1<$ 4#D#4#9A# 3@ 38#14 G?9#4=1GB W138 38# A#$$@I1@8?E4@$)’&
\F19# W@@E 9#B<3@E#UJ#3#A31@9 @D @D 38# O=>?@A :;’// & ’ ( ) *6"