Part:BBa_K4515004
thl-opt
thl-opt
Profile
Name: thl-opt
Base Pairs: 1179 bp
Origin: Clostridium acetylbutyrate ATCC824, genome
Properties: acetyl-CoA acetyltransferase
Usage and Biology
BBa_K4515004 is an enzyme named thl, which encodes acetyl-CoA acetyltransferase and converts Acetyl-CoA into Acetoacetyl-CoA in the N-butanol biosynthesis pathway [1-2]. It has been found that N-butanol can be synthesized naturally in Clostridium, but the tolerance of Clostridium bacteria is not good enough for large-scale production [3-5]. Recently, Lactobacillus Brevis with better N-butanol tolerance has been isolated by researchers. In order to improve the yield of N-butanol, we optimized the codon of the thl gene and inserted the optimized thl DNA fragment downstream of the Pthl promoter (Figure 1), and transformed it into L. brevis ATCC367 competent cells to complete the N-butanol metabolic pathway [6].
Profile
Name: Pthl promoter
Base Pairs: 375 bp
Origin: Clostridium acetylbutyrate ATCC824, genome
Properties: promoter of the thl gene
Usage and Biology
Pthl promoter is a constitutive promoter amplified from the ATCC824 genome which is upstream of the gene thl and it regulates the expression of the downstream genes. In this project, we employed this promoter to regulate the transcription of the thl gene.
Reference
1.Li, J., Zhao, J. B., Zhao, M., Yang, Y. L., Jiang, W. H., & Yang, S. (2010). Screening and characterization of butanol-tolerant micro-organisms. Letters in applied microbiology, 50(4), 373–379. https://doi.org/10.1111/j.1472-765X.2010.02808.x
2.Berezina, O. V., Zakharova, N. V., Brandt, A., Yarotsky, S. V., Schwarz, W. H., & Zverlov, V. V. (2010). Reconstructing the clostridial n-butanol metabolic pathway in Lactobacillus brevis. Applied microbiology and biotechnology, 87(2), 635–646. https://doi.org/10.1007/s00253-010-2480-z
3.Inui M, Suda M, Kimura S, Yasuda K, Suzuki H, Toda H, Yamamoto S, Okino S, Suzuki N, Yukawa H (2008) Expression of Clostridium acetobutylicum butanol synthetic genes in Escherichia coli. Appl Microbiol Biot 77:1305–1316. https://doi.org/10.1007/s00253-007-1257-5
4.Mitchell WJ (1998) Physiology of carbohydrate to solvent conversion by Clostridia. In: Poole RK (ed) Advances in Microbial Physiology, vol 39. pp 31–130
5.Bowles LK, Ellefson WL (1985) Effects of butanol on Clostridium-acetobutylicum. Appl Environ Microb 50:1165–1170
6.Biswas I, Jha JK, Fromm N. (2008) Shuttle expression plasmids for genetic studies in Streptococcus mutans. Microbiology (Reading). Aug;154(Pt 8):2275-2282. doi: 10.1099/mic.0.2008/019265-0.
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]
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