Difference between revisions of "Part:BBa K2739008"

Latest revision as of 16:14, 14 October 2018

Bktb

BtkB is paralogous to the phaA protein, which is involved in the bioplastic Polyhydroxyalkanoate (PHA) synthetic pathway. The PHA pathway is usually conducted through the PHA operon, which involves 3 enzymatic reactions. PhaA catalyses the first step of the pathway and allow the formation of acetoacetyl-CoA through condensation of two molecules of acetyl-CoA. BktB was isolated from R. eutropha H16, and instead of forming acetoacetyl-CoA, it allows the formation of 3-ketovaleryl-CoA and contribute to the formation of PHBV as the end product. The sequence of present Bktb is isolated from Ralstonia Eutropha and codon optimised to E.coli.

Usage and Biology

BktB's main function is to form the 3-ketovaleryl-CoA through condensation of two molecules of acetyl-CoA. BktB was isolated from R. eutrophaH16 as the most important paralogous gene for PHBV production since it showed higher substrate specificity to the C5 monomer and used 3-ketovaleryl-CoA more efficiently (Mifune et al., 2010). The kinetic properties analysis of phaA and BktB revealed that the specific activity of PhaA is 150-fold lower than BktB if 3-ketovaleryl-CoA was used as substrates, and the ability of PhaA and BktB to incorporate 3HV fraction during the production of PHBV differed when growing in M9 minimal medium with 1% glucose and different amount propionate (Slater et al., 1998).This result was further supported by analysing and comparing bktB mutant strain with wild type R. eutropha, which indicated that BktB was responsible for generating 3HV in PHBV.

This part is not submitted but the sequence is used to develop the construct BBa K2739009 (Hybrid promoter-phaCAB-Bktb) and BBa K27390010 (Hybrid promoter-phaCB-Bktb), which allow the further investigation of Bktb's function.

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
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 831

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2739008 short</partinfo>
-BtkB is paralogous to the phaA protein, which is involved in the bioplastic Polyhydroxyalkanoate (PHA) synthetic pathway. The PHA pathway is usually conducted through the PHA operon, which involve 3 enzymatic reactions. PhaA catalyses the first step of the pathway and allow the formation of acetoacetyl-CoA through condensation of two molecules of acetyl-CoA. The bktB was isolated from R. eutropha H16, and instead of forming acetoacetyl-CoA, it allows the formation of 3-ketovaleryl-CoA and contribute to the formation of PHBV end product.
+BtkB is paralogous to the phaA protein, which is involved in the bioplastic Polyhydroxyalkanoate (PHA) synthetic pathway. The PHA pathway is usually conducted through the PHA operon, which involves 3 enzymatic reactions. PhaA catalyses the first step of the pathway and allow the formation of acetoacetyl-CoA through condensation of two molecules of acetyl-CoA. BktB was isolated from R. eutropha H16, and instead of forming acetoacetyl-CoA, it allows the formation of 3-ketovaleryl-CoA and contribute to the formation of PHBV as the end product. The sequence of present Bktb is isolated from Ralstonia Eutropha and codon optimised to E.coli.
@@ Line 9: / Line 9: @@
 ===Usage and Biology===
-BktB's main function is to form the 3-ketovaleryl-CoA through condensation of two moleculesof acetyl-CoA. BktB was isolatedfrom R. eutrophaH16 as the most importantparalogous genefor PHBV production since itshowed higher substrate specificity to the C5 monomer and used 3-ketovaleryl-CoA more efficiently (Mifune et al 2010). The kinetic properties analysis of phaA and BktB revealed that the specific activity of PhaA is 150 fold lowerthan BktBif 3-ketovaleryl-CoA was used as substrates, and the abilityof PhaA and BktB to incorporate 3HV fraction in produced PHBV differed when growing in M9 minimal medium with 1% glucose and different amount propionate(Slater et al., 1998).This result was further supported by themselves analysing and comparing bktBmutant strainwith wild type R. eutropha, which indicated BktB was responsible for generating 3HV in PHBV(ibid).
+BktB's main function is to form the 3-ketovaleryl-CoA through condensation of two molecules of acetyl-CoA. BktB was isolated from R. eutrophaH16 as the most important paralogous gene for PHBV production since it showed higher substrate specificity to the C5 monomer and used 3-ketovaleryl-CoA more efficiently (Mifune et al., 2010). The kinetic properties analysis of phaA and BktB revealed that the specific activity of PhaA is 150-fold lower than BktB if 3-ketovaleryl-CoA was used as substrates, and the ability of PhaA and BktB to incorporate 3HV fraction during the production of PHBV differed when growing in M9 minimal medium with 1% glucose and different amount propionate (Slater et al., 1998).This result was further supported by analysing and comparing bktB mutant strain with wild type R. eutropha, which indicated that BktB was responsible for generating 3HV in PHBV.
+This part is not submitted but the sequence is used to develop the construct BBa K2739009 (Hybrid promoter-phaCAB-Bktb) and BBa K27390010 (Hybrid promoter-phaCB-Bktb), which allow the further investigation of Bktb's function.