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Fermentation regulation of DHA production by Schizochytrium sp.based on osmotic pressure

渗透压调控对裂殖壶菌发酵产DHA的影响



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2012
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ChineseJournalofBioprocessEngineering
Vol.10No.6
Nov.2012
doi:10.3969/j.issn.1672-3678.2012.06.005
STUM
:2011-12-30
OGV!

ÿbdó)*MoûimTãäèéêë
(NCET090157);
fPèÿbçèéêë
(123014);
ìíîgï6gN_°êë
(JH0930);
ìíîÆhg ûiêë
(2008D63)
WX.Y

p
 
b
(1977—),
q

NrÓÅû

üý£¤A

£¤"ö

ÓA–5-

Ï
 
/

– û
),
ÿ!
,Email:biotech@njut.edu.cn
PQRSBžTUVz^f
DHA
‰Ÿ 
m
 
n
1,
o
 
p
1,
qrr
2,
stu
1,
v
 
w

(1
*+1-./ 78>PQ12/3 x=0/12YF@ABCD

*+
210009;
2
y*z{/3

y*
232001)
7
 
i

±²Ì³´µ¶·¸
(0、10、20、30
S
40g/LNaCl)
Ö¹º»¬
HX 308
«¦&
DHA
Ѽ½®¾‰ÇØ
Ù

ÚÛܛ
:20g/LNaCl
û(‡ð¹º»¬k¿S
DHA
ÀÁ

kÂî

ü½®Äî
、DHA
&îÑ
DHA
ÅkÂ
îÇL‹þÿª
73g/L、107g/L、50g/L
S
68mg/g,

DHA
âü½®rÆÅÇþLûK

ª
452%。
Ȃ

âAµ¶É
(10g/LNaCl)
./Ê


40mmol/L
ÌÍ®ÎÏÐ
,DHA
&î­ÑËÒLbKÌ
2821%;
âKµ
¶É
(40g/LNaCl)
./ÊË
40mmol/L
ÓÔ½
,DHA
&îbKÌ
4684%;
ܛËÝîǂBÒÕ¯*íú
(•Ö×8¹º»¬ÀÁ
DHA。
jkl

¹º»¬
;DHA;
µ¶É

ÒÕ¯*í
mnopq
:Q815.1    
6rsFt
:A    
6u?q
:1672-3678(2012)06-0024-05
FermentationregulationofDHAproductionby
Schizochytriumspbasedonosmoticpressure
ZENGYan1,FENGYun1,TONGQianqian2,RENLujing1,HUANGHe1
(1.StateKeyLaboratoryofMaterialsOrientedChemicalEngineering,ColegeofBiotechnologyandPharmaceuticalEngineering,
NanjingUniversityofTechnology,Nanjing210009,China;2.HuainanNormalUniversity,Huainan232001,China)
Abstract:Theefectsofdiferentosmoticpressures(0,10,20,30and40g/LNaCl)onDHAaccumula
tionandfatyacidscompositionofSchizochytriumspHX308werestudied.20g/Lsalinitywasproved
tobebeneficialforcelgrowthandDHAproduction.Celdryweight(DCW),totalcontentoffaty
acids,DHAyield,andDHAconcentrationonDCW werethehighest,around73g/L,107g/L,
50g/L,and68mg/g,respectively.TheDHApercentageoftotalfatyacidsalsoreachedthemaxi
mumof452%.Inaddition,undertheconditionsof40mmol/Lglycinebetaineadditionunderlowsa
linity(10g/LNaCl)and40mmol/Ltrehaloseadditionunderhighsalinity(40g/LNaCl)DHAyield
couldincreaseby2821% and4684%,respectively,thusdemonstratingthepositiveefectofcom
patiblesoluteadditiontobeefectiveforDHAproductionbySchizochytriumsp
Keywords:Schizochytriumsp;docosahexaenoicacid;osmoticpressure;compatiblesolutes
   
u ¶ u ‹ Æ d «
(docosahexenoicacid,
22∶6Δ4,7,10,13,16,19,

DHA),


tÏç
”,
3ω 3
ݧ€u/ày«

ñO®vg×k

Lw­N


x/EyzA{Ö"ê[:žŸÚ
[1]。
opqB
voj

5-:#u

§€u/ày«Jg

–X4
‚gNA6
DHA
ÊÆYA6B-.
[2]。
ijknÓ
A–YA‹/A‚[:žŸrs

S/tu3X.Œ
J|¦Yijl˖Hxl`®¯Yijð/}k

/µZ^8WXYijð`N

oÁ;†Pù‹O&
ñ:A–-U
[3]。
oGijl˖VË4

ͦG
”,
ë75^YͦG
Á

½«

]
Pro)、
~GN3–

]€À»

/E§R8Ž§
R4

]N*

N8

ç‚8/O4
)。Jakobsen
A
[4]
£¤5^€À»#opqBBh
S8(ATCC
20889)
Y+,ͦG
  
¤ñOÿŸ˜7#Y£¤+,ÅUOBÁ
ab
[5]、
<Ÿlm
[6-7]
/¿v¦–HƒL
[8-9]
A"
ê

¡)U

¤+,”ijkT

ª9o
pqB
HX 308
O€tij¼½¾
DHA
ƒ„/à
y«¨X­©

DO˜þ9ƒ„tu—á^¸Í
¦GHX 308
ng

•ij¼
½YµZGEnÌ5-GYrs

1 
vw%xy
11 
vw
111 
Bh
  
opqB
(Schizochytriumsp)HX 308(CCTCC
M209059),
¤ñO_£(y+öaNò

YÉO
UãˆÝ„…–YÉUó
(CCTCC)。
112 
„…
  
Á¦„…˜X1Ï)2
[8],
5-„…±
Tˆ‰4HJä³
120g/L,
çÁJ
10%,
„…CD
4
25℃、160r/min。
113 
¢=
  
ù{N½«€À»/ùú4

Ög…
),
†‡N
Ö:ŀ

12 
^Ws@…
  
®¯*HJ

àH¡¢/ày«eLN1Ï)
2
[8],
šÍA«
(GC)
vw
DHA
>ày«JD
[¶ˆ"«eL
(C19∶0
eL

%4QŒJ¯Œ
DHA
YJ
[10]。
2 
‹Œ%)
21 
PQXYžTUVz
HX 308
^f
DHA
£Z‰Ÿ 
  
opqB
HX 308
5-6
DHA
YG

ÏÐ
Iày«
(TFAs)
J
、DHA
6J
、DHA
>A–J

(mg/g)。
^

4€tij¼½nopqB
HX 308
5-6
DHA
Yrs

$^

S_

ij
¼½nopqB5-6
DHA
YGrs~h

ÌU
20g/LNaCl
3:f&®¯A‚/
DHA
ƒ
„

˜§A–J

Iày«J
、DHA
6J*l˜
3h%

v¥#
73、107
/
50g/L。
ÐãàïS
y6
,DHA
>BWA–JY²ãŽ(h9¢ï¾
}Y+ð

3g4
68mg/g。
opqB6
DHA
¾
,²~µUYijkWX
,^
8ijkug


r€f&opqB5-6
DHA。
n
1 
§¨PQXYžTUVz
HX 308
^f
DHA
‰Ÿ 
Fig.1 EfectsofosmoticpressureonDHAaccumulation
ofSchizochytriumspHX308
22 
PQXYžTUVz
HX 308
[\¢]‰
Ÿ 
  
nÓA–q‰

5-uâUijkY`Nn
ày«YvÔ#ÃF],Y
[11]。
€tij¼½
¾
,SchizochytriumspHX 308
5-6–U+,à
y«YvÔ­©B
1。
$B

S/y6

u/à

C14∶0
/
C16∶0
YJ ¡ijkY¢gq
¢g
;C20∶5n 3
J€ ijk`Nq`N


¡YT€`

Þ>
18%
;<
;C22∶5n 6
J 
¡ijkY9¢q¾}

q
DHA
J ijkY
¢g(^h9¢ï¾}Y+ð

”
0g/LNaCl
º
³Y
411%
9¢Ã
20g/LNaCl
º³¾Y
452%,
åï¾}Ã
40g/LNaCl
º³¾Y
391%,
ؘ
20
g/LNaCl
3µ&opqBƒ„
DHA,
S/O˜C
D¾Nò3MYày«˜X

”_£‘œxy

ày«YvÔ­© ¡ijkY`Nq‹OP
h

ijk5g5:f&u/ày«Yƒ„

q5
€f&‚l§€u/ày«Yƒ„

n&opq
Bxw

ijk(•Ž(gr€f&
DHA
OIà
y«UJY¡g

52 
!

# p
 
bA

ijklmnopqB5-6
DHA
Yrs
¡1 
§¨PQXY±TUVz
HX 308
‰[\¢]
Table1 FatyacidcompositionofSchizochytriumspHX308culturedunderdiferentsalinity(n=3)
ρ(NaCl)/(g·L-1)
w(
ày«
)/%
C14∶0 C16∶0 C20∶5n 3 C22∶5n 6 C22∶6n 3
Ìf
0 66±020 228±053 18±016 134±023 411±108 143±021
10 81±011 238±040 19±021 123±019 440±076 99±009
20 83±015 239±039 17±018 122±020 452±074 87±012
30 91±009 246±042 17±022 122±025 427±055 97±019
40 102±013 259±049 20±012 119±026 391±062 109±016
23 
^_0­Ö`£»ÃžTUVz^f
DHA
£Z‰Ÿ 
231 
—á^¸N½«€À»nopqB5-6
DHA
GYrs
  
O•i
(10g/LNaCl)
5-„…U—á€tä
³YN½«€À»
(0、10、40、70
/
100mmol/L),
ª
9ÌnopqB
HX 308
YijYZœ/EnÌ
5-6
DHA
GYrs

‘œ^
2。
$^

S/y
6

O•i
10g/LNaCl
§

N½«€À»Y—ánA
–Jl:Cžrs

jIày«J
,DHA
6J/E
DHA
>A–JY²%r ¡—áY^¸N½«€À
»Yáá(^h¢gï}•Y+ð

Î¥#—á
40
mmol/L
N½«€À»§
,DHA
6J/
DHA
>A–
JY²rl˜3h%

v¥#
50g/L
/
74mg/g),
²
‘—á§v¥¡g™
2821%
/
2131%。
B


tijCD¾
,^
¸N½«€À»Y—ánopqB
ày«¨XYrs

$B

S_

u/ày«
C14∶
0、C16∶0
J ¡N½«€À»—áJY¢gq}
•

q
DHA
OIày«Uñ>Šv²(^h¢gï
}•Y+ð

O—á
40mmol/L
N½«€À»§l˜
3h%
(498%)。
©9

—á
40mmol/L^
¸N½«
€À»3µ3opqBA‚/
DHA
ƒ„

˜^

°.}ª9™Ogi
(40g/LNaCl)
5-
„…U

—á€tä³YN½«€À»
(0、10、20
/
40mmol/L)
nopqB
HX 308
5-6
DHA
G
Yrs

â•iCD¾‘œÍ‹

giCD¾N
½«€À»Y—ánA–Jl:Cžrs
,DHA
6
J/
DHA
>A–JY²%r(^h¢gï}•Y


£—á
20mmol/L
N½«€À»§
,DHA
6
J/
DHA
>A–JY²v¥#
47g/L
/
69mg/g,
²‘—á§v¥¡g™
2368%
/
2545%;

DHA
OIày«Uñ>Šv²¡g
˜
422%。
9Œ‘œBC

O•i/giCD¾

—
áµJͦG(40
/
20mmol/L)
:f&éÓA–F|€µYWX

”q:f&ëQ

DHA
Yƒ„

n
2 
§¨PQRÀÁ±

0­ab¢cde‰^_žTUVz
HX 308
‰Ÿ 
Fig.2 Efectofexogenousglycinebetaineadditionontheperformanceof
SchizochytriumspHX308under10g/Land40g/Lsalinity(n=3)
62
A
 
–
 
á
 
Ö
 
u
 
â
  
!
10
"
 
¡2 
§¨PQRÀÁ±

0­ab¢cde‰^_žTUVz[\¢]‰Ÿ 
Table2 Fatyacidcomposition(% TFAs)ofSchizochytriumspHX308culturedatdiferentexogenous
glycinebetaineconcentrationsat10g/Land40g/Lsalinity
ρ(NaCl)/
(g·L-1)
c(
N½«€À»
)/
(mmol·L-1)
w(
ày«
)/%
C14∶0 C16∶0 C20∶5n 3 C22∶5n 6 C22∶6n 3
Ìf
10
0 85±023 240±057 15±005 119±023 441±087 100±007
10 71±017 215±068 12±010 145±027 478±056 79±021
40 51±014 199±039 16±009 162±019 498±048 74±013
70 51±020 195±024 20±013 160±022 490±066 84±006
100 52±011 190±033 26±007 158±017 481±059 93±011
40
0 109±018 259±053 18±012 126±013 397±077 91±005
10 99±012 249±067 18±009 134±015 413±068 87±009
20 80±017 253±058 19±013 137±017 422±071 89±012
40 82±025 277±043 16±008 124±016 391±069 110±023
 
232 
—á^¸ùú4nopqB5-6
DHA
G
Yrs
  
ùú4#Ê^.Á^¸Í¦G,^
¸—á˜
„…UéÓA–F|€µYA‚WX


i
(10g/LNaCl)
5-„…U—á€tä³Yùú
4
(0、10、40、80、160
/
200mmol/L),
ª9Ìnop
qB
HX 308
5-6
DHA
GYrs

‘œ^
3。
$^

S_

 ¡ùú4䳔

áá˜
200
mmol/L,
A–J”
65g/L
¡g˜
81g/L。10mmol/L
/
40mmol/L
ùú4n
DHA
ƒ„l:žŸrs

j
£Ì䳡g˜
80mmol/L
§
,DHA
6J”
4g/L
¡

75g/L。
˜^

£—á
40mmol/L
ùú4§

DHA
OIày«Uñ>Šv²3h
(474%)。
ؘ
O•iCD¾—á^¸ùú4€û°opqB


k:n¡g™
DHA
6J

  
˜^k°.}ª9™Ogi
(40g/LNaCl)
5
-„…U
,^
¸—á€tä³Yùú4
(0、10、40
/
80mmol/L)
nopqB
HX 308
5-6
DHA
G
Yrs

‘œ]^

ñO

$^

S/y6

—á
ùú4ïA–JŽ:¡g
,DHA
6J/
DHA
>A
–JY²%r(^6h9¢ï¾}Y+ð

£—
á
40mmol/L
ùú4§
,DHA
6J/
DHA
>A–
JY²%*l˜3h%

v¥#
56g/L
/
75
mg/g,
²‘—á§v¥¡g™
4684%
/
3229%。
B

4€tij¼½CD¾
,^
¸ùú4Y—án
opqBày«¨XYrs

$B

S_

—á
40
mmol/L^
¸ùú4#giCD¾3µ3opqB
6
DHA
Y

˜§u/ày«
C14∶0、C16∶0
J~
•
,DHA
OIày«Uñ>Šv²3g
(454%)。
wCOgiWX¾

—áµJY^¸Í¦Gú4ŽtH:f&éÓA–F|€µYWX

”
q:f&ëQ6–
DHA
Yƒ„

n
3 
§¨PQRÀÁ±

0­fge‰^_žTUVz
HX 308
‰Ÿ 
Fig.3 EfectsofexogenoustrehaloseadditiononperformanceofSchizochytriumspHX308
under10g/Land40g/Lsalinity(n=3)
72 
!

# p
 
bA

ijklmnopqB5-6
DHA
Yrs
¡3 
§¨PQRÀÁ±

0­fge‰^_žTUVz[\¢]‰Ÿ 
Table3 Fatyacidcomposition(% TFAs)ofSchizochytriumspHX308culturedatdiferent
exogenoustrehaloseconcentrationsunder10g/Land40g/Lsalinity
ρ(NaCl)/
(g·L-1)
c(
ùú4
)/
(mmol·L-1)
w(
ày«
)/%
C14∶0 C16∶0 C20∶5n 3 C22∶5n 6 C22∶6n 3
Ìf
10
0 99±017 244±059 15±011 119±017 441±089 82±012
10 95±026 223±078 16±009 123±014 448±059 95±016
40 96±013 213±061 18±005 126±009 474±042 73±020
80 107±025 232±072 13±010 122±012 450±053 76±013
160 110±020 225±069 14±008 121±017 432±046 98±015
200 117±019 215±066 14±012 115±010 417±057 122±012
40
0 100±010 247±051 20±017 116±014 390±070 127±009
10 96±022 220±079 17±010 119±016 441±069 101±013
40 98±016 223±057 19±010 129±008 454±045 77±018
80 102±021 235±062 14±005 128±011 442±059 79±016
 
3 
‹
 
)
ª9opqB
HX 308
O€tij¼½CD
¾Y5-­©ï5^
,20g/LNaCl
º³3:f&o
pqBA‚/
DHA
ƒ„

A–J

Iày«J

DHA
6JE
DHA
>A–JY²rl˜3h%

v
¥4
73g/L、107g/L、50g/L
/
68mg/g。
˜^

k5^^¸Í¦G
N½«€À»

ùú4


:néopqB®¯F|€µYijkWX

”ql˜ëQ6–
DHA
gƒ„YëY

]O•
i
(10g/LNaCl)
CD^¸—á
40mmol/L
N½«
€À»
,DHA
6J/
DHA
>A–JY²*²‘—
á§v¥¡g™
2821%
/
2131%;
Ogi
(40g/LNaCl)
CD¾^¸—á
40mmol/L
ùú4
§
,DHA
6J/
DHA
>A–JY²*²‘—á§
v¥¡g™
4684%
/
3229%。
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