Difference between revisions of "Part:BBa K4515006"

 
 
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crt-opt
 
crt-opt
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== Profile ==
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Name: crt-opt
 +
 +
Base Pairs: 786 bp
 +
 +
Origin: Clostridium acetylbutyrate ATCC824, genome
 +
 +
Properties: 3-Hydroxybutyryl-CoA dehydratase
 +
== Usage and Biology ==
 +
BBa_K4515006 is an enzyme named crt, which encodes 3-hydroxybutyrate CoA dehydrase in host cells, and plays a key role in the metabolic pathway of N-butanol synthesis [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 crt gene and inserted the optimized DNA fragment into the pLY15 plasmid under the regulation of the Pcrt promoter (Figure 1), and transformed it into L. brevis ATCC367 competent cells to form the N-butanol metabolic pathway [6].
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[[File:T--East China--BBa K4515006-Figure1.png|500px|thumb|center|Figure 1. the plasmid map of the composite part BBa_K4515006.]]
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== Profile ==
 +
Name: Pcrt promoter
 +
 +
Base Pairs: 187 bp
 +
 +
Origin: Clostridium acetylbutyrate ATCC824, genome
 +
 +
Properties: promoter of the crt gene
 +
== Usage and Biology ==
 +
Pcrt promoter is a constitutive promoter amplified from the ATCC824 genome which is upstream of the gene crt and it regulates the expression of the downstream genes.
 +
 +
 +
==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.
  
  

Latest revision as of 07:34, 26 September 2022


crt-opt

crt-opt

Profile

Name: crt-opt

Base Pairs: 786 bp

Origin: Clostridium acetylbutyrate ATCC824, genome

Properties: 3-Hydroxybutyryl-CoA dehydratase

Usage and Biology

BBa_K4515006 is an enzyme named crt, which encodes 3-hydroxybutyrate CoA dehydrase in host cells, and plays a key role in the metabolic pathway of N-butanol synthesis [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 crt gene and inserted the optimized DNA fragment into the pLY15 plasmid under the regulation of the Pcrt promoter (Figure 1), and transformed it into L. brevis ATCC367 competent cells to form the N-butanol metabolic pathway [6].

Figure 1. the plasmid map of the composite part BBa_K4515006.

Profile

Name: Pcrt promoter

Base Pairs: 187 bp

Origin: Clostridium acetylbutyrate ATCC824, genome

Properties: promoter of the crt gene

Usage and Biology

Pcrt promoter is a constitutive promoter amplified from the ATCC824 genome which is upstream of the gene crt and it regulates the expression of the downstream genes.


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]