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[硫酸锌对米根霉产L-乳酸的影响]

[Effect of ZnSO4 on L-lactic acid production by Rhizopus oryzae].

作者信息

Ge Chunmei, Pan Renrui, Zhang Jie, Cai Jingmin, Yu Zengliang

机构信息

Department of Biology & Environment Engineering, Hefei College, Hefei 230022, China.

出版信息

Wei Sheng Wu Xue Bao. 2013 May 4;53(5):515-20.

Abstract

OBJECTIVE

To improve the yield and quality of L-lactic acid by Rhizopus oryzae, we aim to understand the relationship between inorganic salts utilization and the L-lactic acid metabolism of the strain RLC41-6, through systematic analysis of the effects of zinc ion concentration on the production of L-lactic acid and the Lactic Dehydrogenase (LDH) activity.

METHODS

Rhizopus oryzae was cultured at 36 degrees C for 36h with different quantity of ZnSO4 in fermentation medium. The fermentation products were monitored by reversed-phase high performance liquid chromatography (RP-HPLC), LDH isoenzyme composition in the cell was analysed by non-denatured polyacrylamide gel electrophoresis (PAGE).

RESULTS

Our results showed that the concentration of ZnSO4 in medium could modulate the expression of LDH isoenzyme except LDH1, especially stimulated the expression of LDH4 and LDH5. When initial concentration of ZnSO4 is above 0.02%, the LDH4 and LDH5 reached the highest level. However, the activity of LDH was inhibited by higher concentration zinc ion in extracellular environment. When ZnSO4 concentration is 0.02%, LDH activity reaches its maximum 200U/mL, the HPLC assay showed only L-lactic in the fermentation products (137 g/L), while the conversion rate of glucose to lactic acid is 91%.

CONCLUSION

Zinc ion can regulate the metabolic processes of Rhizopus oryzae and modulate the types of the final fermentation products. An optimal concentration of ZnSO4 can not only facilitate the LDH expression but also prevent pyruvate from transformation into the malic acid and fumaric acid during the metabolism process, thereby enhance the metabolism of glucose to lactic acid of Rhizopus oryzae.

摘要

目的

为提高米根霉生产L-乳酸的产量和质量,通过系统分析锌离子浓度对L-乳酸产量及乳酸脱氢酶(LDH)活性的影响,旨在了解米根霉菌株RLC41-6对无机盐的利用与L-乳酸代谢之间的关系。

方法

在发酵培养基中添加不同量的ZnSO4,于36℃培养米根霉36小时。采用反相高效液相色谱(RP-HPLC)监测发酵产物,通过非变性聚丙烯酰胺凝胶电泳(PAGE)分析细胞内LDH同工酶组成。

结果

结果表明,培养基中ZnSO4浓度可调节除LDH1外的LDH同工酶表达,尤其能刺激LDH4和LDH5的表达。当ZnSO4初始浓度高于0.02%时,LDH4和LDH5达到最高水平。然而,细胞外环境中较高浓度的锌离子会抑制LDH活性。当ZnSO4浓度为0.02%时,LDH活性达到最大值200U/mL,HPLC分析显示发酵产物中仅含L-乳酸(137g/L),此时葡萄糖转化为乳酸的转化率为91%。

结论

锌离子可调节米根霉的代谢过程并影响最终发酵产物的类型。最佳浓度的ZnSO4不仅能促进LDH表达,还可在代谢过程中阻止丙酮酸转化为苹果酸和富马酸,从而增强米根霉将葡萄糖代谢为乳酸的能力。

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