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Adsorption and catalytic activity of porcine pancreatic lipase on surface modification SBA-15 mesoporous material

表面改性SBA-15材料固载猪胰脂肪酶



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ChineseJournalofBioprocessEngineering
Vol.10No.2
Mar.2012
doi:10.3969/j.issn.1672-3678.2012.02.009
NOPH
:2011-06-29
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IR)
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2/g,
6՘4Â
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SBA 15
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Õ

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SBA 15
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、pH
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:1672-3678(2012)02-0040-05
Adsorptionandcatalyticactivityofporcinepancreaticlipaseon
surfacemodificationSBA15mesoporousmaterial
WANGWei,ZHUKai,YANGBo,HUJicheng,HANPingfang
(ColegeofBiotechnologyandPharmaceuticalEngineering,NanjingUniversityofTechnology,Nanjing210009,China)
Abstract:MediaforenzymeimmobilizationwerepreparedbymodifyingmesoporoussilicaSBA15with
γchloropropyltriethoxysilane(CPTES)group.ThepreparedsupportwascharacterizedwithFTIRand
BETmethodandexhibitedbyhighspecificsurfaceareaanduniformporesize.Furthermore,unmodified
SBA15andthemodifiedSBAClmaterialswereusedforporcinepancreaslipaseimmobilization.There
sultswerecomparedwiththatofenzymeactivity,activityretentionandstability.ThemodifiedSBA15
materialwithCPTESshowedgoodperformancesforporcinepancreaslipaseimmobilization.Compared
withunmodifiedSBA15,theadsorptioncapacityofSBACltoporcinepancreaslipasewasimproved
morethan60%.
Keywords:enzyme;immobilization;mesoporous;molecularsieves;SBA15
  
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W»~¸
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/_>?á
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â¼uP
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7,-
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P
[5]
xÃ[ Û7
SBA 15
†f½ñ
JƸ
SBA 15
‡ˆ‰ŠZ‹}_>?à7•e
 ²”v

ÒãP
[6]
ñ?[

mƒ„…v7
SBA 15,
-]»b+n7䫅v†fƒ]@[
A¡n
Candidarugosa
7Z‹}
(CRL)
‡”F>?
D7µÞ
。Zou
P
[7]
¸²Â/“7åx…v
SBA 15,
qM»bÎOR.n

I•"7
086nm
‚æ3
046nm,
}•ï[à[!O­

„…¸²q`
CPTES
A
SBA 15
ÎGƒ„…
v

6#}n»báIæñƒ„@

16M"‡ˆ
‰ŠZ‹}

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1 
op%rs
11 
op%æç
  
‰ŠZ‹}
(pocinepancreaslipase,PPL),
–—
9ºƒæ/XÕá>ÎçRÇÈ
;PluronicP123
(EO20PO70EO20)、èé@、è(@fY…、s@,Ü
ì1
;NaHCO3、NaH2PO4、Na2HPO4、NaOH,ÑÒê
§ÌëFOÇ
;DSHZ 300A
c©ªFGÅ

ìʓ
—¡ƒhÇ
;85 1
c@eÙÚÅ

Ííbõƒh
Ç
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c
pH
–”2

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È
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cÆ?

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12 
Uop
SBA 15
‚Øّ܅w‰`
  
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SBA 15
7ghañŠ¯
[5]。
  
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SBA 15
W»»Ãƒ„±

S
20g
îï²7W»†f
SBA 15
kŒª=µ~ðk
50mL
ED¢s7Ÿ
˜V1¸
N2±,©Ð20g>?¼¬F—˜γ <
1pè(äp¼¬
(CPTES)
3±M½“úV

S
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N2±7xy=373Kˆ½q48h
*

Sj²©1~†`
30mL
ED(4

¢s

ED
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:

sM37‡“Ê^ܚ¥æðšV
n˜°™

M3ì²
CPTES
ƒ„…v7W»†f

ÿ)&
SBA Cl。
13 
Šß¨ü;£‚ØÙ
  
‡”F}7ghñËañŠ¯
[8]。
±²/`
‡v;U¼Éʖ—ñ˖”jÉÊíu

±²/
Vyz7ÉÊuH$
SBA 15
†f±s‡”7}
ÉÊu

‡”F7}ÉÊ ˆ“u7ñ-I6o
Pa=(ρi-ρf)V/mÖÂM@。‚V:Pa&‘Yˆ“
±‡”7}ÉÊ7u
(mg/g),ρiw ρf&½q,*}
Ÿ/7‚u\]
(g/L),V
&½q/“@
(mL);m
&
q`7ˆ“‚u
(g)。
14 
Šß¨‘ŽYü;££k‚ß
  
k
100mL
7èéçVг
2g
è(@fY…

50mL
£ÂD
、10mLpH
&
70
701/

@eÙ
ÚÅÙÚ
10min
gh9Oò/

®Ÿ/
pH
²”*
HóÎÏ
pH
ª

1гþZ‹}<‡”F7Z
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½q
30min,` 002mol/L
7
NaOH
Ÿ/©
”

q
pH
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HóY*7
NaOH
7u

O
`}•
(U)
Ӈ&k
40℃
7xy=
,30min
U°¥
1μmol(@sFN7}u。
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60℃
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ç)*

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3

$ƒ„…v,w…v*7
SBA 15
7
FT IR
Ã3

o3

I6º@
:SBA 15
ìô7
FT IR
Ã3k
3442cm-1
=@]
Si—OH
ó}Fr
S

k
Si—O—Si
AÿwWAÿó}FrS@]k
470、1092
w
800cm-1。1633cm-1
=7S&
SBA
15
U7NùDFrS
。SBA Cl
7
FT IR
Ã3

Z[jìô7]2S6i

k
604cm-1
=>O`?
57
C—Cl
.7ó}FrS

êƒf
Cl
pT9{õ
ų[
SBA 15
ƒ„
[9]。
f
1 SBA 15
‰`%W‰`op‚’0“:
Fig.1 FTIRspectraofunmodifiedSBA15and
modifiedSBA15materials
  
3

&
SBA 15
 
SBA ClN2¹àªà
(BET)
PªÁ

o3



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LangmuirIV
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ƒ„…v
SBA 15
†f‡ˆ‰ŠZ‹}
c¹àÀÁ

ŒJ÷cW»X‚¹àÀÁ

AJ
SBA 15,
½AŽek
07
23ç
,N2¹àu˜‚
Ð

@][øù±Í

k
SBA Cl
.ɽAŽe
=l3[
05
23

Qç/7š“¹àuñ
SBA 15
>?féyn

ê6f»Ãúû±9[
SBA 15
»bw»T7yn

 
SBA 15
½ñ

ƒ
„…v*7
SBA Cl
†fj»b »T>fé7
=l

kñƒ„@˜>7W7±Í

ƒ
1),
Iã`J
…v7X‚T9{ų~¸ƒ„1k»ÃV*
9åx

IkIæ[ñƒ„@
[10]。
f
2 SBA 15
‰`%W‰`‚
BET
”Ž±
Fig.2 N2adsorptiondesorptionisothermsofunmodified
SBA15andmodifiedSBACl
…
1 SBA 15
‰`%W‰`op‚
BET
…‘yz
Table1 BETresultsofunmodifiedSBA15and
modifiedSBAClmaterials
—­
ñƒ„@

(m2·g-1)
B»b

nm
»“@

(cm3·g-1)
SBA 15 4609 705 09571
SBA Cl 5120 371 06282
 
22 
+ā£Šß¨‚ƒ„
  SBA 15
&¼pW»~¸

jƒ„7¼“p
w}~ÂVÍp@üõV7>?öp1tN¸²
-.ÎGVx

Z[-.i

‡ˆ²³Vˆ“ }
~Â)«¸²9øØew叽ý%`eÎG¹
à

ÂD%`>ÙJ¹à
,¸
²ˆ“ƒ„7_>
SiO2äÛMÃ7}ÂD˜$7½ý%`
[11]。
3

½þ[k
pH
&
70
7s@01Ÿ/7%`=

½qç)AZ‹}7•v7GH

I3

I6º
@

ÀÁ4]@Ö±Í*=l7/0
,SBA 15

ˆç)&
4h
çñ}•¯+à
(218U/g),
k
SBA Cl
k ‡ ˆ
3 h
ç

ñ } • + à

¯ 3
3535U/g。
)*

â㇈ç)wu

•vâ)=
l

•L$‡ˆIB7%`qM»ÃMÃU7}þ
ªåˆ3[‡ˆ}/U
,´
б‡ˆxy¯W3+
û7EK

â㇈ç)7.u

*#}/ð-@
H2Sš²,LM}TÊKŠ‹[。
f
3 
Šß¨+ā
SBA 15
‰`%W‰`op
Š‹Œü;£–£k‚ƒ„
Fig.3 EfectsoftimeonactivitiesofimmobilizedPPL
onunmodifiedSBA15andmodifiedSBACl
f
4 pH

SBA 15
‰`%W‰`opŠ
‹Œü;£–£k‚ƒ„
Fig.4 EfectsofpHontheactivitiesofimmobilizedPPL
onunmodifiedSBA15andmodifiedSBACl
23 pH
£Šß¨‚ƒ„
  pH
A}7•e>÷æ7GH

LM‡ˆWQ
pH
xy=

m
SBA 15
A‡ˆ}•e7GH

M
Þß3
4。
o3



âãIB
pH
7‚æ
,2
m
ˆ“‡ˆ7}ñ•e1‚æ

ák
pH
&

ç¯3


â*=Ò=l

ë…v7
SBA 15
kÿ3ò
~v7IBç

}•0?±Í

k²~7IBVþ
¼Ò=l

k…v*7
SBA Cl
kò~7IBV¯
3+æ7}•

I3

«I6º@

GHè(@f
Y…DE7þ}w‡”FZ‹}½A•v7
pH
k
50~90
9:U
。SBA 15
‡ˆ}w
SBA Cl
‡ˆ}+æ•v1k
pH
&
80
=

ê rgˆ“
p6ÅƂÂò+d
pH
ñòd÷1W½Q
[12]
>
¼

}7Àqv« }7‡”FñË

ˆ“MÃ7
24
/
 
X
 
Ð
 
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³
  
!
10
"
 
v‚wˆ“ƒ„s?!u>¼

ˆ“MÃ7Ð"
%`AJ[à}7Àqvï>÷ªN7%`
[13]。
k
pH
&
90
701/V

Z‹}7½A•vñ?
m

Ip }7Ã2.F¨îÍp@¯p±7!
~Wö>¼

24 
Šß¨£‚sß`
  
6þ}%Añ

‡ˆ

m‡”ˆ“A‡”F
}§²”v7GH

MÞß3
5。
o3



âã
kàªIB=ç)7wu
,3
mۍ7}ñ•e1
4]@=l/0

þ}Žkàª
(60℃)6h
*
Ž£[•v
;SBA 15
‡ˆ}ì
6h
7ઍŽ*
–>ÎÏ}•e7
385%,
½AJ
SBA Cl
>
ÎÏ}•7
391%,
˜†Wæ

;$
SBA Cl
‡ˆ
}•ÏÑàJþ}w
SBA 15
‡ˆ}7}•

ï
N$6
,SBA Cl
‡ˆ}kàª=I6–#à7
}•e

V–•L&‡”FZ‹}ŽkJW»†
f»ÃU

ª]7.FWT
ª]
7ÍàSqZ‹}¹à3W»†f»ÃU»±

q
MÃ2#&²”
[14]。
W»†f7»ÃI6â-}
~Âwx#$kIBV

}I»ÃV0?°¥

q
M‡”}7§²”vM3[[à

f
5 
DŽ
SBA 15
‰`%W‰`opŠ
‹Œü;£–£k‚ƒ„
Fig.5 Efectsoftemperatureontheactivityof
immobilizedPPLonunmodifiedSBA15
andmodifiedSBACl
25 
Šß¨£‚Ssß`
  
‡”F}ª]q`vAJîÿq`(ÓªN

Çj$kO€q`V

3

é8[

m†f‡ˆ}
kº:ª]q`*7}ñ•e.F%&

o3

I
ã
:SBA Cl
‡ˆ}k!

:ª]q`*

ü˜I6
–
121U/g
7ñ}•

k
SBA 15
‡ˆ}!–
="[
41U/g
7ñ}•

‡”FZ‹}7ŒF•
vâãª]:œ7‚Ðk=l

V–•L&W»
†f»»±7¼“p }~Â))$¸²57~
Â)%`e¹à

첺:7½q

~w¡§

q
M}~Â÷T,SBA Cl
7»b½
AJ
SBA 15
7»bNn

ïNf”[}~ÂWì
T<Ž

s6–[½A?à7}•e

f
6 
•¦"M
SBA 15
‰`%W‰`op
Š‹Œü;£–£k‚ƒ„
Fig.6 Efectsofreusecyclesontheactivityofthe
immobilizedPPLonunmodifiedSBA15
andmodifiedSBACl
3 
y
 
)
  
`
CPTES
&ƒ„…v˜

A
SBA 15
ÎGƒ„
…v
,Cl
p1T9{õų3
SBA 15
7ƒ„

ƒ
„…v
SBA Cl
†fñƒ„@?à

&
512m2/g,
»7å“@$
06282m3/g,
»b&
371nm。
ƒ
„…v*»b

»T1>O”³]7}n

;$ñ
ƒ„@‚æ

}•M3[[à

kák
pH
7Àqv
wQ%²”vñ„ïñë…v7
SBA 15
ƒ]N
D

jgh7‡ˆ‰ŠZ‹}>?à7}•ew?
à7î¡IQ%v

+àñ}•k‡ˆIB
pH
&
80
‡ˆ
3h
ç

&
384U/g。
…v†f‡ˆ}k
60

IBV>iD7§²”v

k
6h
*ü˜>
132
U/g
7ñ}•

kª]q`vñ„
,5
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7ñ}•

L˜6j

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