Part:BBa_K1432001
A Synthetic microcystin-degrading Mlr A gene
A Synthetic microcystin-degrading Mlr A gene from Sphingomonas sp. ACM-3962 which codon were optimized using Lactococcus lactis abundant codon.
MlrA is the only microcystin-degrading gene, which cut the Microcystin LR cycle structure to a line.
MlrA is the most important enzyme for the microcystin metabolism mechanism because the cyclic structure promotes microcystin stability against other protease and chemical factors. As the mlrA gene has a very rare sequence and there are almost no homologous genes in the DNA database, we could not predict its origin or family. Since there are no other reports of an mlrA homologous gene, we have very limited genetic information on the biodegradation of microcystin.
In 1994, Jones et.al. isolated a microcystin-degrading bacterium, MJPV,from Australian water bodies for the ¢rst time. The strain was identi¢ed as Sphingomonas sp. based on 16S rRNA gene sequence. Bourne et al. performed cloning and gene library screening of the Sphingomonas sp. strain and detected the microcystin-degrading gene cluster, mlrA,B, C and D. The enzyme encoded by the mlrA gene can cleave the ADDA^Arg peptide bond in microcystin LR and open the cyclic structure. After opening the cyclic structure, linear microcystin LR is degraded by the peptidases encoded by mlrB and mlrC, and divided into each amino acid. The mlrD encodes the transporter protein that allows the uptake of microcystins into the cell.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Sequence of Synthetic mlr A: ATGCGAGAATTTGTTCGACAACGACCATTATTATGTTTTTATGTTTTAGCTATTTTAATTGCTTTAGCTGCTCATGCTTTACGAGCTATGTCACCAACTCCATTAGATCCAATGTTTAAAATGTTACAAGAAACTCATGCTCATTTAAATATTATTACTGCTGTTCGATCAACTTTTGAATATCCAGGAGCTTATACTTTATTATTATTTCCAGCTGCTCCAATGTTTGCTGCTTTAATTGCTACTGGAATTGGATATGGACAAGCTGGATTTCGAGAATTATTATCACGATGTGCTCCATGGCGATCACCAGTTTCATGGCGACAAGGAGTTACTGTTATTGCTGTTTGTTTTTTAGCTTTTTTTGCTTTAACTGGAATTATGTGGGTTCAAACTTATTTATATGCTCCACCAGGAACTTTAGATCGAACTTTTTTACGATATGGATCAGATCCAGTTGCTATTTATGTTATGTTAGCTGCTTCATTATTATTATCACCAGGACCATTATTAGAAGAATTAGGATGGCGAGGATTTGCTTTACCACAATTATTAAAAAAATTTGATCCATTAACTGCTGCTGTTATTTTAGGAATTATGTGGTGGGCTTGGCATTTACCACGAGATTTACCAACTTTATTTTCAGGAGCTCCAGGAGCTGCTTGGTCAGTTATTGTTAAACAATTAGTTATTACTCCAGGATTTATTGCTTCAACTATTATTGCTGTTTTTGTTTGTAATAAATTAGGAGGATCAATGTGGGGAGGAGTTTTAACTCATGCTATTCATAATGAATTAGGAGTTAATGTTACTGCTGAATGGGCTCCAACTGTTGCTGGATTAGGATGGCGACCATGGGATTTAATTGAATTTGCTGTTGCTATTGGATTAGTTTTAATTTGTGGACGATCATTAGGAGCTGCTTCACCAGATAATGCTCGATTAGCTTGGGGAAATGTTCCACCAAAATTACCAGGAGGAGTTGGAGATAAATCAGGAGCTAATGCTTAA
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