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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2019, Vol. 39 Issue (8): 25-31    DOI: 10.13523/j.cb.20190804
Orginal Article     
Enhanced Cyclic Adenosine Monophosphate Production by Coupling Addition of Low-Polyphosphate and Hypoxanthine
CHEN Bao-feng1,2,LI Zhi-gang1,2,ZHANG Zhong-hua2,CHANG Jing-ling1,2,**()
1 School of Life Science and Technology, Henan Institute of Science and Technology, Xinxiang 453003, China
2 Collaborative Innovation Center of Modem Biological Breeding of Henan Province, Xinxiang 453003, China
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Abstract  

In live cells, cyclic adenosine monophosphate (cAMP) was produced from ATP catalyzed by adenylate cyclase and continuous supply of energy and precursor substance was necessary for ATP biosynthesis. The addition of hypoxanthine improved cAMP productivity by 39.1% due to the activated purine salvage pathway when compared with control. However, fermentation ceased at 51h with low cAMP yield and cell concentrations caused by insufficient energy supply. When hypoxanthine and 2g/L-broth (NaPO3)6 were added collaboratively, cAMP concentration (7.24g/L) was improved by 125.5% and 93.5% respectively, compared with those with hypoxanthine and (NaPO3)6 addition individually. The fermentation process with hypoxanthine and (NaPO3)6 coupling addition combined the advantage of salvage pathway and low-polyphosphate which accelerated cAMP biosynthesis and accumulation.

Key words： Cyclic adenosine monophosphate      Hypoxanthine      Low-polyphosphate      Salvage pathway     
Received: 16 November 2018      Published: 18 September 2019
ZTFLH:  Q819  
Corresponding Authors: Jing-ling CHANG     E-mail: changjl001@126.com
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Bao-feng CHEN
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Cite this article:

CHEN Bao-feng,LI Zhi-gang,ZHANG Zhong-hua,CHANG Jing-ling. Enhanced Cyclic Adenosine Monophosphate Production by Coupling Addition of Low-Polyphosphate and Hypoxanthine. China Biotechnology, 2019, 39(8): 25-31.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20190804     OR     https://manu60.magtech.com.cn/biotech/Y2019/V39/I8/25

低聚磷酸盐 添加量(g/L-broth) 添加时间(h)
0 18 30 42
Na5P3O10(3P) 1 1.72±0.03 1.87±0.05 1.95±0.06 1.79±0.05
2 1.80±0.03 2.21±0.06 2.32±0.06 2.04±0.06
3 1.88±0.04 2.10±0.03 2.37±0.03 2.02±0.04
(NaPO3)6(6P) 1 1.80±0.05 1.98±0.03 2.43±0.06 1.96±0.03
2 1.92±0.02 2.57±0.04 2.80±0.02 2.40±0.02
3 1.85±0.03 2.05±0.05 2.52±0.03 2.42±0.03
对照 0 1.73±0.02 — — —
Table 1 The effects of low-polyphosphates on cAMP production with different addition occasions and amounts
Fig.1 The effect of hypoxanthine addition amounts on cAMP biosynthesis
Fig.2 The time courses of cell, glucose hypoxanthine concentrations and specific rate during batch fermentations with or without hypoxanthine addition (a)■/□: OD600;●/○:Glucose concentrations (b) ▲/△:cAMP concentration;◆/◇:Hypoxanthine concentration Solid symbols: Control; Blank symbols: With 1g/L-broth hypoxanthine added at 26h and 37h, respectively (c),(d)——:Control;- - -:With 1g/L-broth hypoxanthine added at 26h and 37h, respectively
Fig.3 The time courses of cell, glucose hypoxanthine concentrations and specific rate during batch fermentations with (NaPO3)6 addition (a)■/□:OD600;●/○:Glucose concentrations ( b)▲/△:cAMP concentration;◆/◇:Hypoxanthine concentration Solid symbols:With 2g/L-broth (NaPO3)6 added at 24h; Blank symbols:With 2g/L-broth (NaPO3)6 added at 24h and 1g/L-broth hypoxanthine added at 26h, 37h and 46h, respectively ( c),(d)——: With 2g/L-broth (NaPO3)6 added at 24h; - - -:With 2g/L-broth (NaPO3)6 added at 24h and 1g/L-broth hypoxanthine added at 26h, 37h and 46h, respectively
发酵批次(#) 发酵时间(h) 最大OD600 葡萄糖消耗量(g/L) cAMP(g/L) 发酵强度[g/(L·h)]
对照 72 1.37 56 3.32 0.046
HX 51 1.23 46 3.21 0.063
24h 6p 60 1.62 55 3.74 0.062
24h 6p+HX 72 1.18 54 7.24 0.101
0h 6p 56 1.54 58 3.58 0.064
0h 6p+HX 67 1.34 53 6.32 0.094
Table 2 Performance comparison for different cAMP fermentation modes
Fig.4 The schematic diagram for cAMP biosynthesis with hypoxanthine and (NaPO3)6 coupling addition PRPP: Phosphoribosyl pyrophosphate; IMP: Inosinate; SAMP: Adenylosuccinic acid
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