Abstract
Objectives
To engineer a small nonconding RNA anti41 to enhance nisin yield by inhibiting the expression of glnR in Lactococcus lactis F44.
Results
We constructed a screening library to determine appropriate artificial sRNAs and obtained a sRNA anti41 that can produce approximately three fold of the inhibitory effect on GlnR. Moreover, the transcription levels of the direct inhibitory targets of GlnR (glnP, glnQ, amtB, and glnK) were dramatically upregulated in the anti41 overexpression strain (F44-anti41), thereby confirming the inhibitory effect of anti41 on GlnR. In addition, anti41 overexpression improved the survival rate of cells by approximately three fold under acid stress, promoted cell growth, and increased nisin yield by 29.83%.
Conclusions
We were able to provide a novel strategy for the construction of robust high-producing industrial strains.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cheigh CI, Pyun YR (2005) Nisin biosynthesis and its properties. Biotechnol Lett 27:1641–1648
Fernandez M, Zuniga M (2006) Amino acid catabolic pathways of lactic acid bacteria. Crit Rev Microbiol 32:155–183. https://doi.org/10.1080/10408410600880643
Gunka K, Commichau FM (2012) Control of glutamate homeostasis in Bacillus subtilis: a complex interplay between ammonium assimilation, glutamate biosynthesis and degradation. Mol Microbiol 85:213–224. https://doi.org/10.1111/j.1365-2958.2012.08105.x
Jian Z, Caiyin Q, Feng W, Zhao X, Qiao B, Zhao G, Qiao J (2016) Enhance nisin yield via improving acid-tolerant capability of Lactococcus lactis F44. Sci Rep 6:27973
Jin Y, Wu J, Li Y, Cai Z, Huang JD (2013) Modification of the RpoS network with a synthetic small RNA. Nucleic Acids Res 41:8332–8340
Komasa M, Fujishima K, Hiraoka K, Shinhara A, Lee BS, Tomita M, Kanai A (2011) A screening system for artificial small RNAs that inhibit the growth of Escherichia coli. J Biochem 150:289–294
Larsen R, Kloosterman TG, Kok J, Kuipers OP (2006) GlnR-mediated regulation of nitrogen metabolism in Lactococcus lactis. J Bacteriol 188:4978–4982. https://doi.org/10.1128/JB.00025-06
Miflin BJ, Habash DZ (2002) Carbon and nitrogen relationships and signalling. The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops. J Exp Bot 53:979
Muro-Pastor MI, Florencio FJ (2003) Regulation of ammonium assimilation in cyanobacteria. Plant Physiol Biochem 41:595–603. https://doi.org/10.1016/s0981-9428(03)00066-4
Qi J, Caiyin Q, Wu H, Tian K, Wang B, Li Y, Qiao J (2017) The novel sRNA s015 improves nisin yield by increasing acid tolerance of Lactococcus lactis F44. Appl Microbiol Biotechnol 101:1–11
Shin JM, Gwak JW, Kamarajan P, Fenno JC, Rickard AH, Kapila YL (2016) Biomedical applications of nisin. J Appl Microbiol 120:1449–1465
Storz G, Vogel J, Wassarman KM (2011) Regulation by small RNAs in bacteria: expanding frontiers. Mol Cell 43:880–891
van der Meulen SB, de Jong A, Kok J (2016) Transcriptome landscape of Lactococcus lactis reveals many novel RNAs including a small regulatory RNA involved in carbon uptake and metabolism. RNA Biol 13:353–366. https://doi.org/10.1080/15476286.2016.1146855
Wang Y et al (2015) Identification of a novel small non-coding RNA modulating the intracellular survival of Brucella melitensis. Front Microbiol 6:164
Wright PR et al (2014) CopraRNA and IntaRNA: predicting small RNA targets, networks and interaction domains. Nucleic Acids Res 42:119–123
Wu H et al (2018) Contribution of YthA, a PspC family transcriptional regulator to Lactococcus lactis F44 acid tolerance and nisin yield: a transcriptomic approach. Appl Environ Microbiol. https://doi.org/10.1128/AEM.02483-17
Acknowledgements
This study was supported by the National Key R&D Program of China (2017YFD0201405), the National Natural Science Foundation of China (31270142, 31770076, and 31570089), and the Funds for Creative Research Groups of China (21621004). Dr. Jianjun Qiao was supported by The New Century Outstanding Talent Support Program, Education Ministry of China.
Supporting information
Table S1—Oligonucleotides used in this work.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Miao, S., Wu, H., Zhao, Y. et al. Enhancing nisin yield by engineering a small noncodding RNA anti41 and inhibiting the expression of glnR in Lactococcus lactis F44. Biotechnol Lett 40, 941–948 (2018). https://doi.org/10.1007/s10529-018-2550-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-018-2550-3