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Effect of biotin and cerulenin addition on DHA production by Schizochytrium sp.

生物素和浅蓝菌素添加对裂殖壶菌发酵产DHA的影响



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ChineseJournalofBioprocessEngineering
Vol.10No.1
Jan.2012
doi:10.3969/j.issn.1672-3678.2012.01.008
OPQI
:2011-06-09
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.
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:1672-3678(2012)01-0042-04
EffectofbiotinandceruleninadditiononDHAproductionby
Schizochytriumsp
RENLujing,WEIPing,FENGYun,JIXiaojun,HUANGHe
(ColegeofBiotechnologyandPharmaceuticalEngineering,NanjingUniversityofTechnology,Nanjing210009,China)
Abstract:ToimprovetheDHAproductivity,efectsofbiotinandceruleninadditiononDHAproduction
wereinvestigatedbasedonDHAsynthesispathwayofSchizochytriumsp.Theresultsshowedthatthebio
tinadditioncouldenhancethelipidaccumulation,whiletheceruleninadditioncouldincreaseDHAand
polyunsaturatedfatyacidcontent.ThepercentageofDHAinceldryweightreached1126% at
03mg/Lbiotinaddition,thusitwas13% higherthanthatofthecontrol.When01mg/Lcerulenin
wasadded,DHAcontentreached122%.
Keywords:biotin;cerulenin;Schizochytriumsp.;DHA
  
;PC¹3{ê‹

°ÇÕeëeìæí‹
(C22∶6,n-3,DHA)
Õÿ!;¼{rº7îTE^
¦6o

Íγ´iô
DHA
“½ì<=‡W

2
]ïŠ÷6tQšì…ÚØÙþÍ+ìPý;
[1],
ËÙÚ*PÜðñòÒ¦îÒñl

T&

FP
}óµ€m‡ˆ¹º
DHA
Ëúí³´EÞv

7r
„…†Z÷7]€;=]ŸE^¼Ÿ=

Do
CAíÕûö4l-*PTŠE
DHA
€^Z
[2]。
DHA
EŽúâpí

ôƒ÷

!oƒ÷

ÕÕd
eDx¿Dˆ‚

gô‹ìa
(TCA)、
üý‹„‚v
é€^ƒõ
CoA

NADPH[3];
!eƒ÷Ճõ
CoA

NADPH
x¿I;í3véî
PKS
véŽú;
P¹3{ê‹
[4]。
7qgEŽúvéBC
DHA
‡
ˆ¿Àà÷Ú¹

µÎ)Ú
DHA
Žúöà

R"

@AÕ
DHA
ŽúEtËv鴇

x¿
~ƒõKh

ô-hEôQe²€m\)Ú
DHA
ŽúE!oƒ÷

x¿~{ꋎúhED
¹ ‚ƒZ\)Ú!eƒ÷

÷#ÕtËvéE
1Çé)Ú
DHA
‡ˆEÍ.

í2]„…†ZE
¼{Ìٙ÷6¼{r
DHA
Eõ™2GfB

Èí
{ê‹tËX¹E³´ŒµQW

1 
vw%xy
11 
@6
„…†Z
(Schizochytriumsp)HX 308[5],
@A
ғC3ãJù\VÏ<

8ruYscåmïí
ršïí
(CCTCCM209059)。
12 
mnK
¬­cåQ
(g/L):MgCl23、CaCl2·2H2O1、
KH2PO44、ˆœÐ 2、¡¢‹÷ 10、KCl2、NaCl15、
FeCl301、MgSO4·7H2O5、deD 40;121℃、30
min
klÀçZ

©÷²Ìp?í
02%
E™Îcå
QrpÇ~‹€B1、B6、B12。
‡ˆcåQ
(g/L):MgCl23、(NH4)2SO46、
KH2PO44、ˆœÐ 2、¡¢‹÷ 10、KCl2、NaCl15、
FeCl301、MgSO4·7H2O5、deD 100;121℃、
30min
klÀçZ

©÷²Ìp?í
02%
E™Îc
åQrpÇ~‹€B1、B6、B12。
13 
mnxy
¬­cå

Y¯¼Úrï+E¬­RÊùö¬
­cåQEßø$Eg1ý`r½ñ:-

à²Ì
p?í
5%
ER¬™(+cå

t

cåë9
20~
24h,
ݍ
25℃,
f
150r/min。
ý`‡ˆcå

à
10%
ER¬™Y¬­cå•
RʇˆcåQr

ýÎcåóD‰ù

cåݍ
25℃,
f
150r/min。
14 
n¯xy
<=T’™¼{23Í.>çDE
[6];
{
ê‹[)¹º6ºò^¯pq>çDE
[7]。
2 
‚ƒ%„)
21 
¤Å0üž
DHA
Y‰¡U
211 
€m\@<=€;

¼{™
DHA
õ™E
F)
“{ê‹EŽúvér

ƒõKh

ô-hÕ
`¼E{ꋎúh
[8],
!/Y-ƒõKh

f-
íqe‹ãõ
CoA,
Õ(½Ê{ê‹EŽú


*úÕ{ꋎúE“Dû

>Õ{ꋎúE
ó,ғ

€m\’™£@
DHA
‡ˆfrEF)Æi

Òr

8i

âï

<=â÷“‚€m\~E
klÀ0ð€;

nô€m\©¹Õ„…†Z€;
E½‡R­

7“~
03mg/L
€m\V

¼{™
8
2257g/L
45žó
(2887±087)g/L。DHA
š<=T’™Ep?j¿
(1126±032)%,
Ã@
çž6
13%,
êë
DHA
^™8
6g/L
452]ó
(800±022)g/L,
nô~?£E€m\öF
Ü
DHA
EŽú

-¬Â½ìPš€m\ՃõK
h
A、
ô-hEôQe²
[9]
öc<óø

š
BaezSal
dana
þ
[10]
E³´/æoC

®
1 
0ü¤Åþž&%ýþ

ÿ!b
DHA
þ‰¡U
Table1 EfectsofbiotinconcentrationsonparametersinDHAfermentationSchizochytriumsp.HX308
ρ(€m\)/
(mg·L-1)
ρ(<=T’™)/
(g·L-1)
ρ(J¼{)/
(g·L-1) m(DHA)(<=T’™)
-1/%
ρ(DHA)/
(g·L1)

6000±181 2257±067 1000±030 600±015
01 6332±189 2426±073 1027±020 650±012
02 6666±182 2645±080 1044±015 696±020
03 7101±213 2887±087 1126±032 800±022
04 6533±195 2481±074 909±027 594±020
06 6200±185 2260±067 717±021 450±015
212 
€m\@{ê‹pÛEF)
€m\E~BC<={ê‹pۇ€6ôýE
X-

/æ‹Ö
1。


âï

0~
03mg/L
€m


=Ñüý‹
(C14∶0)
¼‹
(C18∶1)
‰ÂÀ,8
9

pÇ8
1194%

043%

807%

013%。
7
eëìþí‹
(EPA)

DHA
õ™8
13%

456%
p
34 
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# yËÍþ

~€m\‚ƒZ\@„…†Z‡ˆ^
DHA
EF)
Ç7]¿
18%

50%,
iô€m\@¹3{ê‹
(PUFAs)
EÌÙöù0E½ìP

m
1 
0ü¤Åþž!ªq‰¡U
Fig.1 Efectsofbiotinconcentrationsonfatyacid
compositionSchizochytriumspHX308
213 
€m\@„…†Z‡ˆ^1ÿíEF)
~€m\@„…†Z‡ˆ^1ÿíEF)/
æ‹Ö
2。


âï

?’™£E€m\~™
(01mg/L),
©o2q1ÿíõ™ý;EX-


™£ž¿
03mg/L
ë

þ€T<=rõ1ÿí
057mg,
òÃ@ç2]6
30%,
1ÿí^™8@ç
E
2668mg/L
2]¿
4098mg/L。
m
2 
0ü¤Åþž«¬­®þ‰¡U
Fig.2 Efectsofbiotinadditiononsqualenecontent
andyieldbySchizochytriumspHX308
22 
¤Å¥¦@ž
DHA
Y‰¡U
221 
‚ƒZ\@<=€;

¼{
DHA
õ™E
F)
‚ƒZ\Õo¬ÛZ8€,
!/D¹{ê‹
ŽúvérEβ 1õ ACPŽúhø1ÿíŽ
úvérE
HMGCoA
Žúh

„…†Z=ÑE3
{ꋎú‡¼x¿{ꋎúhvé

(¹3
{ê‹EÌÙòx¿
PKS
véŽú
[4]。
‚ƒZ\@
DHA
‡ˆEF)Æi

Òr

8
i

âï

‚ƒZ\E~@<=T’™¼{õ
™F):£

70’™£í
01mg/L
ë
,DHA
õ
™8
(600±015)g/L
2]ó
731g/L
45


DHA
š<=T’™Ep?í
(1220±036)%,
ò
@Ü@ç2]6
24%。
(0‚ƒZ\’™£½
   
®
2 
¥¦@ž&0ˆ

­ÿ!¶
DHA
®þ‰¡U
Table2 EfectsofdiferentceruleninconcentrationsonparametersinDHAfermentationbySchizochytriumspHX308
ρ(‚ƒZ\)/
(mg·L-1)
ρ(<=T’™)/
(g·L-1)
ρ(J¼{)/
(g·L-1) m(DHA)(<=T’™)
-1/%
ρ(DHA)/
(g·L-1)

6000±181 2257±067 1000±030 600±015
010 5993±180 2197±066 1220±036 731±021
025 5981±189 2198±056 1030±030 616±018
10 5983±179 2058±061 789±023 472±014
25 5948±078 1975±059 672±020 402±012
50 5777±173 1907±057 640±019 370±011
oD7]¿
50mg/L
ë
,DHA
^™,ó
(370±
011)g/L,
J¼{,í
(1907±057)g/L,
iô]£
E‚ƒZ\@¼{ÌÙ
DHA
Žú¹F

222 
‚ƒZ\@{ê‹pÛEF)
‚ƒZ\E~@{ê‹pÛEF)ÆÖ

Ò
r



âï

~?£E‚ƒZ(01mg/L)
nD¹ü!‹
(C14∶0)
]"‹
(C16∶0)
EŽú

Õ
(2]¹3{ê‹Eõ™
,DHA
’™p?È8
46%
2]¿
51%。
70’™£½oD7]ó

mg/L
ë
,DHA
E’™p?,ó
41%。
~
01mg/L
E‚ƒZ\Y
DHA
E’™p?8
46%
2]¿
51%,

Alen
þ
[11-13]
E³´/æòE

nô‚ƒZ\â
÷öŸÂ½¹3{ê‹EŽú

]£E‚ƒZ
\D¹6C¹3{ê‹
(PUFAs)
6
DHA
EŽú

44
€
 
m
 

 
4
 
¿
 
À
  
!
10
"
 
â!ՂƒZ\“D¹{ꋎúhvéEêë>
p„6
PKS
véEŽú

m
3 
¥¦@ž!ªqnt‰¡U
Fig.3 Efectsofceruleninconcentrationsonfatyacid
compositioninSchizochytriumspHX308
223 
‚ƒZ\@1ÿíõ™EF)


âï

)‚ƒZ\~™E7]

1ÿ
íõ™ÌÍ,?

0‚ƒZ\E’™£í
5mg/L
ë

1ÿíõ™^™8@çE
044mg/g

2668
mg/L
pÇÀ,ó
018mg/g
6
10mg/L。
1ÿíõ
™E,?â!š‚ƒZ\D¹6
HX 308
#wŽú
¿ÀrE
HMGCoA
Žúhöó

R"

‚ƒZ\È
â!n,?
HX 308
<=Ñ}”wEõ™

m
4 
¥¦@¤Åþž«¬­®þ‰¡U
Fig.4 Efectsofceruleninadditiononsqualenecontent
andyieldbySchizochytriumspHX308
3 
‚
 
)
%&6ó,hEKh€m\÷6hED¹ ‚
ƒZ\@
Schizochytriumsp.HX 308
‡ˆ;!EF
)

/æýr

~
03mg/L
E€m\ë
,DHA
š<
=T’™$pÃj
1126%,
ò@Ü@ç2]6
13%;
~
01mg/L
‚ƒZ\ë
,DHA
š<=T’
™$pÃâj
122%,
í
DHA
E‡ˆ

)ځõ™E
2]2G6WEpË

7û²EtËX¹óöνo
D³´



D*PE

¬Í.

@
DHA
Žúö
ôýE½ìP

R"

âù0HP!45$€^]
õ™E
DHA


7%@¿€^ú:í%¯

V#
â*+&)ÚpË

чQí}E)ÚÍ.

M¼6q

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54 
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DHA
EF)