免费文献传递   相关文献

Medium optimization of Phoma putaminum LYYZ90-19 for cellulase production by response surface design

核茎点霉发酵产纤维素酶的培养基优化



全 文 :!10
"!

#
2012
$

%
T
 
L
 
´
 
Y
 
µ
 

ChineseJournalofBioprocessEngineering
Vol.10No.5
Sep.2012
doi:10.3969/j.issn.1672-3678.2012.05.003
NOPH
:2012-02-14
JQR!

·¸

gÞh

»®ijij¾¿ÀÁ
(2008BADC4B08);
»®”·b»®©çÀÁ
(2010DFA62510);
’¼‹P(}kl0hi
j¾¿ÀÁ
(IRT0963);
m“pÅÓT»Å(}ÀÁ
(CX2010B287);
Âno»®p‡AÀÁ
ST.U

qrs
(1987—),
t

m“uv‰

ÑÒÅÓT

ÅÓÔÕ

5¤ÑTL

w
 
x

Ùډ
),
ŽÅÓC
,Email:tianyun79616@163com
ÈÉÊË]ÌÍ­Î^vwJ ¡
fgh
1,2,
i-.
1,
j
 
k
1,2,
lmn
1,2
(1
o.pq61234cde

rs
410128;2
o.q6/0 12F07805

rs
410128)
7
 
a

A+Õ34ìh5Ï|}~
(Phomaputaminum)LYYZ90 19
ÓÑÒã€ÄÅÆâ<=>

ÁKLB
CƂƒ„

NO
Box Behnken
BCDE…^

à€†‡r!Õ3râ<Õ34ìh

ˆ‰
Minitab
Š)ϋŒf
gâ<ìh

kð=>YÓÄÅÆ()

Ž
427g/L,
O
079g/L,K2HPO4059g/L。Ã=>()*Ñґ
6۠
1347U/mL,
’=>l“ñ6€†!
773U/mL,
6€†ïšm”
741%。
bcd

|}~

•–L€

Õ345

ÑÒ
efghi
:TQ92    
6jklm
:A    
6n;i
:1672-3678(2012)05-0013-04
MediumoptimizationofPhomaputaminumLYYZ9019for
celulaseproductionbyresponsesurfacedesign
PANLeyi1,2,YINJiangnan1,TIANYun1,2,LUXiangyang1,2
(1.HunanAgriculturalBioengineeringResearchInstitute,Changsha410128,China;
2.ColegeofBioscienceandBiotechnology,HunanAgriculturalUniversity,Changsha410128,China)
Abstract:TheoptimalmediumofPhomaputaminumLYYZ9019wasstudiedbysinglefactorexperi
mentandresponsesurfacedesignwithMinitabsoftware.Theresultsshowedthattheoptimalmedium
wereasfolows:427g/Lbran,079g/Lpeptoneand059g/LK2HPO4.Undertheconditions,
theenzymeactivitywas1347U/mLItwasincreasedby741%,higherthanthatoftheoriginal
strainof773U/mL.
Keywords:Phomaputaminum;celulase;responsesurfacedesign;ferment
  
y·

CDE¾{z{

È$2‹l]^LX

10
|
t,
YÆCD`]}á;¯|~

éÝ*
¾{ŸW£Dá;žýk²2Å0Pˤ

;€)
DZª/ú‡2a‡

KEQö÷æ;"2Ԃ
ÞÝˤCDEFJKCDE01T>æà¥
/J|7øù:ƒ29îé{

BKé{…†
[1]。
  
º^

mªÄ2YÆF¬„…/*ÞÁQ2
•G

è/–†*ބ…

Š‹Jh‡;ˆH22
‰Š‹}1Lè¼k

(cqný

hm>CDE
F2ŒS
LYYZ90 19,
GžE¼–Ig¯

¤
456²³7-s>FGHId89

J#-C
DEFYÆ9T>,Ž"M

1 
op%rs
11 

  
Sh
LYYZ90 19
›Š‹T÷ˆË¤…¾
²Ïqn7
[2]
(‡;ˆH‰2Š‹}1LcT²
ÏqnTž

h
ITS
¿|{`²³û»è?@A
B
(Phomaputaminum),
G9˜Àm“p‹ÆTL
Y¶ÅÓT

12 
vwJ
  PDA
GHI
(g/L):
‘’
200(
“”•–


bL
25min,
˜¬µ·
),
NO<
20,
¢e‚
12。
  
z*GHI
(g/L):
NO<
1,(NH4)2SO405。
  
01GHI
(g/L):
tu™‚š
25,
1˜,­L
3,NaCl1000,K2HPO4 500,MgSO4·7H2O050,
CaCl205,FeSO4·7H2O075,MnSO4·H2O200。
13 
vwrs
  
­N9˜
4℃
°±›c2
PDA
GHI

œ
20mL
üSbÀGHI…6

žŸ£² Ç²R
;›*2§¨©œl´;
75mL
z*GHI2
250
mL
¡¢³c

ž£¤¥GH
(28℃、140r/min),
ÈÉ
6h
¤‘¦i£§i×z*©c2›*£(

å´µ
ý
108
Dzz*©J,ç²£
5%
2±z(±l0
1GHIc

ž£¤¥GH
4d(28℃、140r/min)[3]。
ÈÅGH[\d

wc¼–

y­c:A

14 
ÏÎÐÑÒ
  
­ž³01
4d
Ç201©
,3000r/min
ÏÐ
5min,¯
’©Wè¨F©
[4]。
15 
ÓÔJÌÍ­ÕÎÖ×
(CMCase)^
ØÙ
  
²ÏÐÇ2F©½L
20
¿

í
05mL
´l
´;p(²£
1%CMC Na
2¼½c
,´
©ªXB
æ©
20mL


Ml
50℃
¬Úb­c<9
30min,
­NÇ´l
DNS
¼
20mL,
®b­cL
®
5min,¯
°Ç»1ý
25mL


¤qz²ÄÄ
µi
540nm
º
OD540。JL®¹_F©è-Û,
ºž
OD
±²NO
³­—<(
[5]。
F
_åG‚»´
(U/mL):1
ÅF_G‚»´è¯µ
[\t
,1mL
F©
1min
}G9ŠL>T
1μmolDŒ<(JNO–ˆ)\¶

wc¼–

16 
`žŸg¨
  
Ó¿
Minitab
Ê\-¼–yd¡i²
³
[6]。
ç‹"#2GžE¼–ÅÓ2µ¶c

g
-01GHId/Œ89

Ä»/GHIc·
–p(´µ

”“¡p(´µP
K2HPO4p(´µ
*

ŞEæ¢bc

²ÝÝ·–
35g/L、
”“¡
06g/L
P
K2HPO4075g/L。+Ig¯,ÂÃ
Box behnken
¼–ªiŒœ
[7],
JF_åè45
A

·–

”“¡P
K2HPO42p(´µè¹„(,
ªi456¼–

2 
xy%z)
21 Box Behnken
¦§xy
  
ˤ
Minitab
Ê\(¤
Box Behnken
¼–ˆ

M¸Í
15
ˆ¼–

¼–ªiryڅ
1。
œ
1 Box Behnken
¦§©ª®xy
Table1 DesignandresultsofBoxBehnkenexperiment
¿Ý
X1
ρ(·–)/
(g·L-1)
X2
ρ(”“¡/
(g·L-1)
X3
ρ(K2HPO4)/
(g·L-1)

°F_

(U·mL-1)
1 -1(25) -1(04) 0(075) 1279
2 1(45) -1 0 1311
3 -1 1(08) 0 1343
4 1 1 0 1308
5 -1 0(06) -1(05) 1353
6 1 0 -1 1337
7 -1 0 1(10) 1302
8 1 0 1 1313
9 0(35) 0 0 1301
10 0 1 -1 1409
11 0 -1 1 1371
12 0 1 1 1332
13 0 0 0 1368
14 0 0 0 1373
15 0 0 0 1357
 
22 
¶"·¸¹ºÚÛ®ÜÝÞg¨
  
ˤ
Minitab
Ê\WÀK-«²³-…


–ydWÔ-«È©

žý/…ßF_å45
A„O¹„(ÊÿÚ2ÁwWÀK-«Ô¶

  Y=136667+00025X1 +01625X2 -
01125X3-04208X

1-01458X

2+00042X

3-
015X1X2+005X1X3-0375X2X3 (1)
  
-K
(1)
dÔ¸²³

yڅ
2。
  



:P=0005<001,
…ðļ–n¤
£~2-«`°±ñ/

£~2È©µPJ¤E
»Ú£
R2
a…ß


Minitab
Ê\¡ižýQ£
~22
R2

09606,
!ð;ŸN
9606%
2>F
41
T
 
L
 
´
 
Y
 
µ
 

  
!
10
"
 
(„À²¬gÅÓ2

ŞEc

…ðQ£~Ô
¶2È©µ±³

œ
2 
¶"ßR๺¯Rr»g¨
Table2 Analysisofvarianceforquadratic
polynomialmodelforoptimization
ofenzymaticactivity
a{
DF SS MS F (P>F)
-«À
9 168417 0187130 1353 0005
$`À
3 031250 0104167 753 0027
cÔÀ
3 070917 0236389 1709 0005
èéç¤
3 066250 0220833 1596 0005
¹¸É¸
5 006917 0013833
ÿÈÀ
3 006250 0020833 625 0141
ƒÉ¸
3 000667 0003333
©i
14 175333
 
  

Minitab
Ê\dž*ñ/`²³P£~
Ô¶2Ô¸²³

yU…





PJä
N

ÞwÀ
X2、X3-F_å234ñ/(P<005),
X1{ñ/;ÁwÀ X12-F_å234mµñ/
(P<001),X22、X32{ñ/;èéÀ X2X3-F_å
234mµñ/
(P<001),X1X2、X1X3{ñ/。£
~2ÿÈÀ
P=0141>005,
ž+-«£~¾©

PJ¤+£~a-
LYYZ90 19
201>Fd²
³P¥º

œ
3 
¶"ßR๺áM®âÜÝÞg¨
Table3 Regressioncoeficientsandtheirsignificance
forquadraticpolynomialmodel
À Ú£ Ú£Qñɸ

֖

…(
136667 006791 201261 00001
X1 00025 004158 0095 0937
X2 01625 004158 3908 0011
X3 -01125 004158 -2705 0043
X21 -04208 006121 -6875 0001
X22 -01458 006121 -2383 0063
X23 00042 006121 0068 0948
X1X2 -01500 005881 -2551 0051
X1X3 00500 005881 0850 0434
X2X3 -0750 005881 -6377 0001
23 
`žŸs ¡vwJ
  
K
(1)
2Ig¯


Minitab
Ê\-sd
456²³

Õ¬NF_å45A„Ož*245
6Pym$

ÚÞ
1~
Þ
3。
--«Ô¶d²³

³sÞº».¼Þº».yÀ
0,
PJžýOž
Eæ¢bcA

²Ýè
X1=0772,X2=0992,X3=
-0628,
W·–

”“¡P
K2HPO42æ¢p(´
µ²ÝÝ
427、079
P
059g/L,
Tž£Ã7lK
(1)
cPž
Y=136U/mL。
  
è/–Ø-«£~2ñÄ`P;x`

·–
427g/L、
”“¡
079g/L、K2HPO4059g/L[td–ؼ–

c:°F_ý/
1347U/mL,
㴼A
(136U/mL)
€°î¸{ý
1%。
^89
"QS°F_è
773U/mL,
°F_,m/N
741%。
ž+£¤4567892>FGHIPÙ
;x

ã;•¤ïA

f
1 
ãäå€%æçèZÎÖ×^_`
Fig.1 Efectsofbranandnentoneon
enzymaticactivity
f
2 K2HPO4qæçèZÎÖ×^_`
Fig.2 EfectsofK2HPO4andpeptoneon
enzymaticactivity
51 
!

# qrsy

?@AB01>CDEF2GHI89
f3 
ãäq
K2HPO4ZÎÖ×^_`
Fig.3 EfectsofbranandK2HPO4on
enzymaticactivity
3 
x
 
)
  
ˤ¡i¼Ô7¤X/?@AB
LYYZ90 19
>CDEF2ÁwWÀK-«£~

@µ-£~2
²³P456²³

-QS2>FGHId/8
9

æ¾GH[(g/L):
·–
427,
”“¡
079,
K2HPO4059。hµ–ؼ–,Q[\tF_å
45Aè
1347U/mL,
ý/¥ºA2
9904%,
     
^89"QS°F_è
773U/mL,
°F_,m/
N
741%。
Øð£¤456²³7-
LYYZ90 19
2>F01I289{©œ
,^
HPÙ;x

L6j

[1] 
#½Á

QpCDEJKF„TL¾¬
[M].
¿À

9¼YÆ
NÁ•
,2011:2324.
[2] 
ÂÃ

ÄÅÆ

ÇÈ

m>CDEFShqnr„no
[J].
c·‹¼@i
,2011,27(24):108111.
[3] 
‹ÕÉ

5JÊ

ËÌ-

y

ŒS‹Q˜Q—›(º»2¾
â9
[J].
c·TL<Í
,2007(23):137141.
[4] 
ËoÎ

ÏÐÑ



y

Þh>CDEFŒS2qn

û»
rF¼`pŒÅÓ
[J].
ÑTL¼–Ã
,2009,29(3):
7276.
[5] 
ÒØÓ

5´Ü

ÔՓ

y

·–-›ÖQB
RutC 30
>C
DEF2edç¤
[J].
TLp9¼Y¶
,2009,43(2):
2733.
[6] MustafaK.Optimizationofmicrowaveassistedextractionproce
dureforzincandcopperdeterminationinfoodsamplesbyBox
Behnkendesign[J].FoodComposAnal,2009,22:343346.
[7] FereiraSLC,BrunsRE,FereiraHS,etal.BoxBehnkende
sign:analternativefortheoptimizationofanalyticalmethods
[J].AnalChimActa,2007,597(2):
櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒櫒
179186.
,-./
éê„ëìí#$ÅÆ4™Ç
c·»¼/n×TLé{„µ¶ÅÓTé{Øf¾{klÙÚcÅÓCÛ-ÑØTLÜlT>ªÄ
PéF34Ÿ2ÑØle,­

ÑØTLÜlG9ytÝÿó®¯d/ڐÅÓ

y0…
《Biore
sourceTechnology》
–ï

hÞß

ÏÐyÔ7.µTž2Øbc—b(m
70%


6¡2le,­]ߥI-àØdá
⬝؂

éàØîá⵶2éFXîµÑØ>N2é{

èJ),léF„ªÄ

QklÅh/a
ãä[\tm—b(Øb2Z±le,­ÅÓ

0GJåÓ

ü02¥/çè&

ÑØle,­x%ý
902%。
ìÇ

dލÅ0/¥/ æ{M&,­®¯

le,­x%ý
90%,
é&¤(ŸIµJ)

„Jç,­Ô7€°

…GN±m2,­x%P±)2éF

QklDÅÓ/àØbè$µ¶cÑØleª²2„9

0G؎h¯éÇ

@#è$#Ǐ}
2:l¡gPG9TªÝÏefX

ŸŸ,mÑØle2Xï

ú‡34Ç#2TLÜlGg9µ¶

-
+

Qkl0h89/Xê9¥g9 ëê9Gg92TLÜlG9YZ

G9x%±Œ
100%,
¬2T
LÜlOÀPQý/TLÜl·¸Qñ
GB/T20828—2007
PìíTLÜl
EN14214



œ+›

61
T
 
L
 
´
 
Y
 
µ
 

  
!
10
"