Composite

Part:BBa_K4907134

Designed by: Jincheng Lyu   Group: iGEM23_XMU-China   (2023-09-16)
Revision as of 01:36, 11 October 2023 by CHENGTT (Talk | contribs)


I0500-B0034-Mn-SOD-T7 promoter-katGE

Biology

Mn-SOD

The partial reduction of oxygen leads to the formation of various reactive oxygen species (ROS). Superoxide anion radical (O2•−) is usually the first ROS to be generated. The efficiency of enzymatic reactions decreases at low temperature, leading to the accumulation of intracellular peroxides and causing damage, which is one of the mechanisms of bacterial cold injury. SODs are the main antioxidant enzyme families in organisms. They are considered the first defense line against oxidative stresses due to their function of converting O2•− to H2O2 and H2O (1,2). Mn-SOD, a subtype of SOD, utilizes manganese (Mn) as a cofactor, existing in the form of a homotetramer. It is located within the mitochondria of aerobic cells. The expression of the Mn-SOD gene in several insect species increased their survival at low temperature (3).

KatG

KatG is a bifunctional enzyme with both catalase and broad-spectrum peroxidase activity. It can convert H2O2 to H2O and O2, which can promote bacterial growth at low temperature to some extent. Furthermore, it also performs NADH oxidase, INH lyase, and isonicotinoyl-NAD synthase activities (1).

T7 RBS

T7 RBS is a ribosome binding site on the genome of the T7 phage, which is capable of recruiting ribosomes in engineered E. coli.

Usage and design

Through previous experiments, we have proved that Mn-SOD (BBa_K4907023) can improve the low temperature stress resistance of engineered bacteria. Mn-SOD can catalyze active oxygen species in bacteria to generate hydrogen peroxide. Considering that hydrogen peroxide is also harmful to bacteria, we hope to introduce KatG as a catalytic enzyme to degrade hydrogen peroxide catalyzed by Mn-SOD. We use BBa_K4907132, BBa_K4907041, No part name specified with partinfo tag. and BBa_K4907132</b></center>

Reference

1. N. B. Ackerman, F. B. Brinkley, Oxygen tensions in normal and ischemic tissues during hyperbaric therapy. Studies in rabbits. Jama 198, 1280-1283 (1966).
2. J. M. McCord, I. Fridovich, SUPEROXIDE-DISMUTASE - THE 1ST 20 YEARS (1968-1988). Free Radical Biology and Medicine 5, 363-369 (1988).
3. Y. I. Kim et al., Modulation of MnSOD protein in response to different experimental stimulation in Hyphantria cunea. Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology 157, 343-350 (2010).
4. B. L. Triggs-Raine, B. W. Doble, M. R. Mulvey, P. A. Sorby, P. C. Loewen, Nucleotide sequence of katG, encoding catalase HPI of Escherichia coli. Journal of Bacteriology 170, 4415-4419 (1988).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2081
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2081
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2081
    Illegal BglII site found at 2116
    Illegal BglII site found at 3051
    Illegal BglII site found at 3178
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2968
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2081
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2081
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


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