Part:BBa_K4515005

hbd-opt

hbd-opt

Profile

Name: hbd-opt

Base Pairs: 849 bp

Origin: Clostridium acetylbutyrate ATCC824, genome

Properties: β-Hydroxybutyryl-CoA dehydrogenase

Usage and Biology

BBa_K4515005 is an enzyme named hbd, which encodes β-Hydroxybutyryl-CoA dehydrogenase and transfers acetoacetyl-CoA into 3- Hydroxybutyryl-CoA 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 hbd gene and fused the optimized hbd DNA fragment to the crt-ter DNA fragment with another RBS2 site and co-transcripted (Figure 1), and transformed it into L. brevis ATCC367 competent cells to complete the N-butanol metabolic pathway [6].

Figure 1. the plasmid map of the composite part BBa_K4515005..

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]