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Screening of chemically defined medium for succinic acid fermentation

发酵法生产丁二酸的化学合成培养基的筛选



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ChineseJournalofBioprocessEngineering
Vol.11No.5
Sep.2013
doi:10.3969/j.issn.1672-3678.2013.05.009
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ActinobacilussuccinogenesNJ113
xœ{“”

*ž“”hiœªhŸ ì²z¾3
Ae’!¥¦§

=Ÿ z¾x89p
(Glu)、
c9p
(Met)
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G¥‘^3Ae’!¥¦§¦F
(g/L):CH3COONa136,NaCl10,MgCl202,CaCl202,
Na2HPO4031,NaH2PO416,KH2PO43,NH4HCO3157,Glu087,Met011,VH0010,VPP0025。X3L{|§Ú
¨©,50g/L
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Screeningofchemicalydefinedmediumforsuccinicacidfermentation
MAJiangfeng,XIYonglan,XURong,DAIWenyu,
CHENKequan,JIANGMin,WEIPing
(StateKeyLaboratoryofMaterialsOrientedChemicalEngineering,ColegeofBiotechnologyand
PharmaceuticalEngineering,NanjingUniversityofTechnology,Nanjing211800,China)
Abstract:AccordingtothegrowthcharacteristicsofActinobacilussuccinogenesNJ113,achemicaly
definedmedium(CDM)wasdesigned,whichcanmaintainthestraintogrowthon.Thekeyfactorsin
theCDMwereGlu,Met,VH andVPP.Withphosphatebufersaltsintheinitialmedium,thefinal
compositionsoftheCDMwereasfolows(g/L):CH3COONa136,NaCl10,MgCl202,CaCl202,
Na2HPO4031,NaH2PO416,KH2PO43,NH4HCO3157,Glu087,Met011,VH0010,VPP0025.
Anaerobicbatchfermentationwascariedoutina3LfermentorintheCDM:Theinitialconcentrationof
glucosewas50g/L,withasuccinicacidconcentrationof452g/Landayieldof904% after70h.
Comparedwiththeinitialmedium,theyieldofsuccinicacidwasincreasedby252%,andthe
byproductwasremarkablydecreased.
Keywords:Actinobacilussuccinogenes;succinicacid;chemicalydefinedmedium;keyfactor;fermentation
  
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3H2O155,pH70。
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(g/L):CH3COONa136,NaCl1,
MgCl202,CaCl202,Na2HPO4031,NaH2PO416,
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ßÓ{}CXW

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Table1 Concentrationofaminoacids
µ8xQ² ρ/(g·L-1) µ8xQ² ρ/(g·L-1)
LAsp 021 LLeu 053
LThr 022 LTyr 011
LSer 024 LPhe 028
LGlu 087 LHis 008
Gly 015 LTyp 009
LAla 043 LLys 021
LVal 032 LArg 022
LMet 011 LPro 011
LIle 028 LCys 020
_
2 
MÞa¬v­®
Table2 Concentrationofvitamins
¥&¦Q² ρ/(mg·L-1) ¥&¦Q² ρ/(mg·L-1)


(VH)
10
ƒx
(VPP)
25
ªx
(FA) 10

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(DVB5)
25
Áx„…ª
(VB6)
50
¥&¦
B12
(VB12)
05
À†¦
(VB1)
25 ´µ8þx(PABA) 25
bǦ
(VB2)
25 À‡x(LA) 25
_
3 
&N”¬v­®
Table3 Concentrationofsalts
9)ÁQ² ρ/(mg·L-1) 9)ÁQ² ρ/(mg·L-1)
µ8B©x
1500 CuSO4·5H2O 10
MgSO4·7H2O 3000 AlK(SO4)2·12H2O 10
MnSO4·H2O 500 H3BO3 10
FeSO4·7H2O 100 Na2MoO4 25
CaCl2·2H2O 100 NiCl2·6H2O 25
CoCl2·6H2O 100 Na2WO4·2H2O 25
ZnCl2 13 NaSeO3 10
14 
DXPQ
141 
’“À…T‹Œ
t
1mL
¾¿ÕÖ{µ¶ÔU‡ð7™&È
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1mL
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25mL
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mL
T
5%
‡
HCl
½
MgCO3,Œ0,™8ÇS”
â⅁Â
660nm
¸‹ŒöâD
(OD660)。
143 
XWx1ÛT‹Œ
ҙ×Ø{ݔ•Õ
(HPLC),
”•”3–

9
:
DionexUltimate3000


”•”ù—
:Altech
Prevail
XWx—

˜™Ý
:25mmol/LKH2PO4,pH
25(
™
H3PO4³±);˜ˆ:1mL/min;)¨2:
HPLC
Kg×Ø{ݔ•–
(WatersHPLC2010

2
);
š‹a

8ǚ‹a

8ǚ‹Úi
:215nm;
µ5Û
:20μL;—„:25℃。
144 
n7xcÙT
n7xcÙ
=m(
n7x
)/m(
ªÀ
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PQ`Órš›lsY
Fig.1 Efectsofmissingfactoronsuccinic
acidfermentation
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2 
óô`Órš›lsY
Fig.2 Efectsofaddingfactoron
succinicacidfermentation
Fµ8xjPF¼ïCú=ç¼ð£ò\V¡T
¨™

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ÏÐ#*j
ÕÖ8¼{}üšµ8xßÓ{¼PQµ8xT
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7

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X’Û’“Àå­

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rX’
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ÂۃT&iCúWXV¡
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F\óϒV>($¾¿TTRµ¶

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žn¾¿Øô

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4。
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10g/L,
~ÕÖ
12h,
è
645¼’“Àçèn·
,OD660zY£23*+;
ó`!
VH7¾¿ØôÝ´ß1,’“ÀHXå­,
OD660Y£12*+,û£º245TPv;` !VPP
7

X
27g/L
’“Àå­
,OD660£132。S,VHá
VPPT`!VQ$ÍA.succinogenesNJ113T&i
n7xT&¢

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FàÓ¢ÕÖ8¼T

Q\óϒ”o£
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VHáVPP。ÈÓu¾¿ÕÖ8¼T”xyzÁ¢”,
®+Á£TFàÓ¢ÕÖ87D
(g/L):CH3COONa
136,NaCl1,MgCl202,CaCl202,Na2HPO4031,
NaH2PO416,KH2PO43,NH4HCO3157,Glu087,Met
011,VH0010,VPP0025。
C
3 
PQRS¡,%“rš›lsY
Fig.3 Efectsofmissingonekindofamino
acidsonsuccinicacidfermentation
C
4 
PQRS¡MÞarš›lsY
Fig.4 Efectsofmissingonekindofvitamins
onsuccinicacidfermentation
23 
u?@`Ól±Ç¢£%rš›²Kx“l
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A.succinogenesNJ113
&i½¯ÑT\ó&iϒ

™FàÕÖ8¨£¾¿ÕÖ8

jªä’“À
50g/L
Ž
,3L
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A.succinogenes
NJ113
j1XFàÓ¢ÕÖ8¼T¾¿fõ

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ۃ&iàˆ
,9h
7±äŽÅ

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44h
7

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17g/L
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7

n7xT§Û…¥_j
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g/L,
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n7xcÙY£
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÷…Tüš

C
5 
T
A.succinogenesNJ113
Uu¢£%l
3L
š›VBÞ²Kx“
Fig.5 SuccinicacidproductionbyA.succinogenes
NJ113usingCDM in3Lfermentor
3 
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8¼T

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º¼\óT¥&¦£
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D
(g/L):CH3COONa136,NaCl1,MgCl202,
CaCl202,Na2HPO4031,NaH2PO416,KH2PO43,
NH4HCO3157,Glu087,Met011,VH0010,VPP
0025。
j
3L
¾¿~,µ¶<¦ö<
,50g/L
ªä
’“À¾¿
70h,
n7xT§Û…£
452g/L,
n
7xcÙY£
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¿>^n7xÝÞ

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GH

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