不同培养方式下dCO<sub>2</sub>对黑曲霉发酵产糖化酶的影响

doi:10.13523/j.cb.20170105

中国生物工程杂志

2017, Vol. 37

Issue (1): 27-37 DOI: 10.13523/j.cb.20170105

研究报告

不同培养方式下dCO₂对黑曲霉发酵产糖化酶的影响

王利群, 鲁洪中, 储炬, 王永红

华东理工大学生物反应器工程国家重点实验室上海 200237

Dissolved Carbon Dioxide Effects on Glucoamylase Synthesis of Aspergillus niger in Batch and Chemostat Cultures

WANG Li-qun, LU Hong-zhong, CHU Ju, WANG Yong-hong

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China

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摘要：

通过在进气中混入部分CO₂气体，在分批培养和恒化培养两种培养方式下研究了不同dCO₂水平对黑曲霉产糖化酶的影响。在分批培养方式下，较高的dCO₂对细胞的生长具有抑制作用，并且随着dCO₂水平的增加，抑制程度加强，但是较高浓度的dCO₂对糖化酶的合成有利。而恒化培养时，低稀释率D₁=0.05/h下，较高的dCO₂对细胞的生长并没有明显的抑制作用，但有利于糖化酶合成。高稀释率D₂=0.08/h下，较高的dCO₂对细胞的生长有明显的抑制作用，并且抑制程度随着dCO₂水平的增加而加大。以上实验结果表明，dCO₂对细胞生长和糖化酶合成的影响不仅和dCO₂水平有关，也和培养方式及细胞所处的特定代谢状态有关。这对于黑曲霉产糖化酶工业放大过程中补料分批发酵中比生长速率的设计具有很好的指导作用。

关键词： 黑曲霉; 糖化酶; 生长抑制; CO₂胁迫

Abstract:

The effect of different levels of elevated dissolved CO₂ availability(CO₂ stress) was investigated in batch and chemostat cultures of Aspergillus niger by gassing with CO₂-enriched air. Under batch cultures, higher elevated dissolved CO₂ availability in culture medium resulted in a more significant decrease in specific growth rate, whereas it appeared to have a positive effect on glucoamylase synthesis. Under chemostat cultures with low dilution rate (D₁=0.05/h), high levels of dissolved CO₂ led to no reduction of specific growth rate, but an increase of glucoamylase production. Whereas at high dilution rate (D₂=0.08/h), both substrate uptake and cell grow were more severely inhibited by more elevated dissolved CO₂. The effect of elevated dissolved CO₂ availability on the fermentation process depends on CO₂ levels as well as dilution rate. Therefore, the influence of carbon dioxide on cell growth and glucoamylase production dependent on CO₂ levels, cultivation mode and specific metabolic activity, which would be helpful for the design of specific growth rate in scale-up of glucoamylase fermentation by Aspergillus niger.

Key words: Aspergillus niger Glucoamylase Growth repression CO₂ stress

收稿日期: 2016-10-25 出版日期: 2017-01-25

ZTFLH:

Q815

基金资助:

国家“973”计划（2013CB733600），国家国际科技合作专项（2013DFG32630）资助项目

通讯作者: 王利群 E-mail: lqwang2013@126.com

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引用本文:

王利群, 鲁洪中, 储炬, 王永红. 不同培养方式下dCO₂对黑曲霉发酵产糖化酶的影响[J]. 中国生物工程杂志, 2017, 37(1): 27-37.

WANG Li-qun, LU Hong-zhong, CHU Ju, WANG Yong-hong. Dissolved Carbon Dioxide Effects on Glucoamylase Synthesis of Aspergillus niger in Batch and Chemostat Cultures. China Biotechnology, 2017, 37(1): 27-37.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170105 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I1/27

[1] 郭艳梅,郑平,孙继宾. 黑曲霉作为细胞工厂——知识准备与技术基础. 生物工程学报,2010, 26(10):1410-1418. Guo Y M, Zheng P, Sun J B. Aspergillus niger as a potential cellular factory:prior knowledge and key technology. Chinese Journal of Biotechnology, 2010, 26(10):1410-1418.
[2] Melzer G, Dalpiaz A, Grote A, et al. Metabolic flux analysis using stoichiometric models for Aspergillus niger:comparison under glucoamylase-producing and non-producing conditions. Journal of Biotechnology, 2007, 132(4):405-417.
[3] Marin N J, Polaina J. Glucoamylases:structural and biotechnological aspects. Applied Microbiology and Biotechnology, 2011, 89(5):1267-1273.
[4] Mhairi M, Brian M. Effects of elevated dissolved CO₂ levels on batch and continuous cultures of Aspergillus niger A60:an evaluation of experimental methods. Aplied and Environmental Microbiology,1997, 63(11):4171-4177.
[5] Song H, Lee J W, Choi S, et al. Effects of dissolved CO₂ levels on the growth of Mannheimia succiniciproducens and succinic acid production. Biotechnology and Bioengineering, 2007, 98(6):1296-1304.
[6] Gros J.B., Dussap C.G., Catté M. Estimation of O2 and CO₂ solubility in microbial culture madia. Biotechnology Progress, 1999, 15(5):923-927.
[7] 赵凯,徐鹏举,谷光烨. 3,5-二硝基水杨酸比色法测定还原糖含量的研究. 食品科学, 2008, 29(8):3-7. Zhao K, Xu. P. J, Gu G Y. Study on determination of reducing sugar content using 3,5-dinitrosalicylic acid method. Food Science, 2008, 29(8):534-536.
[8] Youngquist J T, Lennen R M, Ranatunga D R, et al. Kinetic modeling of free fatty acid production in Escherichia coli based on continuous cultivation of a plasmid free strain. Biotechnology and Bioengineering, 2012, 109(6):1518-1527.
[9] Mcintyre M, Mcneil B. Dissolved carbon dioxide effects on morphology, growth, and citrate production in Aspergillus niger A60. Enzyme and Microbial Technology, 1997, 20(2):135-142.
[10] Mcintyre M, Mcneil B. Effect of carbon dioxide on morphology and product synthesis in chemostat cultures of Aspergillus niger A60. Enzyme and Microbial Technology, 1997, 21(7):479-483.
[11] El-Sabbagh N, Mcneil B, Harvey L M. Dissolved carbon dioxide effects on growth, nutrient consumption, penicillin synthesis and morphology in batch cultures of Penicillium chrysogenum. Enzyme and Microbial Technology, 2006, 39(2):185-190.
[12] Lu S, Eiteman M A, Altman E. Effect of CO₂ on succinate production in dual-phase Escherichia coli fermentations. Journal of Biotechnology, 2009, 143(3):213-223.
[13] Richard L, Guillouet S E, Uribelarrea J L. Quantification of the transient and long-term response of Saccharomyces cerevisiae to carbon dioxide stresses of various intensities. Process Biochemistry, 2014, 49(11):1808-1818.
[14] Aguilera J, Petit T, De Winde J H, et al. Physiological and genome-wide transcriptional responses of Saccharomyces cerevisiae to high carbon dioxide concentrations. FEMS Yeast Research, 2005, 5(6-7):579-593.
[15] Yusaku F, Hiroshi M. Improved glucoamylase production by Rhizo-pus sp.A11 using metal-ion supplemented liquid medium. Journal of Fermentation and Bioengineering, 1996, 82(6):554-557.

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