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寡糖诱导子对青霉素 G 产生菌Penicillium chrysogenum 作用方式的定量 PCR 研究。

Quantitative PCR study on the mode of action of oligosaccharide elicitors on penicillin G production by Penicillium chrysogenum.

机构信息

Applied Biotechnology Research Group, Department of Molecular and Applied Biosciences, University of Westminster, London, UK.

出版信息

J Appl Microbiol. 2009 Oct;107(4):1131-9. doi: 10.1111/j.1365-2672.2009.04293.x. Epub 2009 Apr 9.

Abstract

AIM

To investigate the effects of single and multiple additions of the oligosaccharide elicitors, obtained from alginate and locust bean gum, on penicillin G production and the transcript level of penicillin G biosynthetic genes.

METHODS AND RESULTS

The transcript copy numbers and penicillin G concentration in liquid cultures of Penicillium chrysogenum grown under control and elicited conditions were compared using quantitative PCR and HPLC assay respectively. An increase in the penicillin G production rate and transcript copy numbers of the three major penicillin G biosynthetic genes pcbAB, pcbC and penDE was observed in the elicited cultures compared to control cultures. The effects were observed to be higher in multiple elicitor added cultures compared to single elicitor supplemented and control cultures.

CONCLUSIONS

The results show, for the first time in bioreactor cultures, the enhancement of penicillin G transcript copy number of the penicillin biosynthetic genes using qPCR with a corresponding increase in the penicillin G production upon multiple elicitor addition of two different types of elicitors.

SIGNIFICANCE AND IMPACT OF THE STUDY

Establishment of the effect of multiple elicitor addition on penicillin G production and investigating the role of oligosaccharide elicitors as transcriptional activators has wide spread impact for antibiotic industry.

摘要

目的

研究从褐藻酸钠和槐豆胶中获得的寡糖诱导剂的单次和多次添加对青霉素 G 生产和青霉素 G 生物合成基因转录水平的影响。

方法和结果

分别使用定量 PCR 和 HPLC 测定青霉素 G 生物合成的三个主要基因 pcbAB、pcbC 和 penDE 的转录拷贝数和青霉素 G 浓度在控制和诱导条件下培养的产黄青霉液体培养物中的青霉素 G 产量。与对照培养物相比,在诱导培养物中观察到青霉素 G 生产速率和三个主要青霉素 G 生物合成基因 pcbAB、pcbC 和 penDE 的转录拷贝数增加。与添加单一诱导剂和对照培养物相比,在添加多种诱导剂的培养物中观察到的效果更高。

结论

这些结果首次在生物反应器培养物中表明,使用 qPCR 增强青霉素生物合成基因的青霉素 G 转录拷贝数,并在添加两种不同类型的诱导剂时相应增加青霉素 G 的产量。

意义和研究的影响

确定多次添加诱导剂对青霉素 G 生产的影响,并研究寡糖诱导剂作为转录激活剂的作用,这对抗生素行业具有广泛的影响。

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