Coding

Part:BBa_K2606001

Designed by: Zhengming Zhu   Group: iGEM18_Jiangnan_China   (2018-10-07)


msmK


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 775
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

It is a fragment cloned from Lactococcus lactis NZ9000 gene group, which is verified to be a new and effective anti-acid gene in Lactococcus lactis NZ3900.

Usage and function

The msmK overexpression strain has 213-fold higher survival rate than parent one at pH 4.0 after 3 hours.

T--jiangnan_china--yl--3.png

T--jiangnan_china--yl--4.png

Fig 1 Survival rate at acid stress (pH 4.0)

T--jiangnan_china--wet--9.png

Fig 2 Electron microscopy of L.lactis NZ3900/pNZ8149-msmk-cspD2-gfp and L.lactis NZ3900/pNZ8149 before and after acid stress

Before the acid stress, the cell structure of the control strain and the recombinant strain remained intact. After 3 hours of pH 4.0 stress, the cell membrane thickness became thinner and the surface became rough, and the cell membrane of some control strains ruptured. In comparison, the cell structure of the recombinant strain remains more intact, thereby effectively reducing the damage caused by acid stress on the cells.

Source

msmK is derived from L.lactis NZ9000 and is a genomic DNA gene sequence.

References

1.Nezhad MH, Hussain MA, Britz ML. 2015. Stress responses in probiotic Lactobacillus casei. Crit Rev Food Sci Nutr 55:740-749.

2.Wu CD, Huang J, Zhou RQ. 2014. Progress in engineering acid stress resistance of lactic acid bacteria. Appl Microbiol Biotechnol 98:1055-1063.

3.Zhang J, Caiyin Q, Feng WJ, Zhao XL, Qiao B, Zhao GR, Qiao JJ. 2016. Enhance nisin yield via improving acid-tolerant capability of Lactococcus lactis F44. Sci Rep 6:12.

4.Martani F, Berterame NM, Branduardi P. 2017. Microbial stress: From molecules to systems (Sitges, November 2015). New Biotechnology 35:30-34.

5.Bosma EF, Forster J, Nielsen AT. 2017. Lactobacilli and pediococci as versatile cell factories - Evaluation of strain properties and genetic tools. Biotechnol Adv 35:419-442.

6.Teusink B, Bachmann H, Molenaar D. 2011. Systems biology of lactic acid bacteria: a critical review. Microbial Cell Factories 10:17.

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