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Inhibition of Coenzyme Qs Accumulation in Engineered Escherichia coli by High Concentration of Farnesyl Diphosphate.

Samoudi M, Omid Yeganeh N, Shahbani Zahiri H, Shariati P, Hajhosseini R - Avicenna J Med Biotechnol (2015 Jul-Sep)

Bottom Line: Over-expression of ispA under the control of stronger trc promoter, however, led to a severe decrease in CoQ 10 production in both recombinant E. coli Ba and E. coli Br strains, as reflected by reductions from 629±40 to 30±13 and 564±28 to 80±14 μg/g Dried Cell Weight (DCW), respectively.The results showed high level of FPP reduces endogenous CoQ 8 production as well and that CoQs are produced in a complimentary manner, as the increase in production of one decreases the production of the other.The reduction in CoQ 10 production can be a result of Dds inhibition by high FPP concentration.

View Article: PubMed Central - PubMed

Affiliation: Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.

ABSTRACT

Background: Coenzyme Q 10 (CoQ 10 ) is an isoprenoid component used widely in nutraceutical industries. Farnesyl diphosphate synthase (FPPS) is a responsible enzyme for biosynthesis of farnesyl diphosphate (FPP), a key precursor for CoQs production. This research involved investigating the effect of FPPS over-expression on CoQs production in engineered CoQ 10 -producing Escherichia coli (E. coli).

Methods: Two CoQ 10 -producing strains, as referred to E. coli Ba and E. coli Br, were transformed by the encoding gene for FPPS (ispA) under the control of either the trc or P BAD promoters.

Results: Over-expression of ispA under the control of P BAD promoter led to a relative increase in CoQ 10 production only in recombinant E. coli Br although induction by arabinose resulted in partial reduction of CoQ 10 production in both recombinant E. coli Ba and E. coli Br strains. Over-expression of ispA under the control of stronger trc promoter, however, led to a severe decrease in CoQ 10 production in both recombinant E. coli Ba and E. coli Br strains, as reflected by reductions from 629±40 to 30±13 and 564±28 to 80±14 μg/g Dried Cell Weight (DCW), respectively. The results showed high level of FPP reduces endogenous CoQ 8 production as well and that CoQs are produced in a complimentary manner, as the increase in production of one decreases the production of the other.

Conclusion: The reduction in CoQ 10 production can be a result of Dds inhibition by high FPP concentration. Therefore, more effort is needed to verify the role of intermediate metabolite concentration and to optimize production of CoQ 10 .

No MeSH data available.


Related in: MedlinePlus

No significant changes were observed in growth associated with ispA over-expression. Error bars indicate the standard error of the mean of three independent experiments.
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Figure 8: No significant changes were observed in growth associated with ispA over-expression. Error bars indicate the standard error of the mean of three independent experiments.

Mentions: In addition, it was observed that the reduction of CoQ10production in both E. coli BaTi and E. coli BrTi occurred simultaneously with an increase in CoQ8production when compared to E. coli Ba and E. coli Br (from 360 to 890 μg/g DCW in E. coli BaTi and 246 to 580 μg/g DCW in E. coli BrTi). However, induction by IPTG (0.05 mM) decreased the CoQ8biosynthesis as well in both E. coli BaTi (to 750 μg/g DCW) and E. coli BrTi (to 310 μg/g DCW) as shown in figures 6 and 7. The growth of recombinant strains was also studied and showed no significance changes by ispA over-expression (Figure 8).


Inhibition of Coenzyme Qs Accumulation in Engineered Escherichia coli by High Concentration of Farnesyl Diphosphate.

Samoudi M, Omid Yeganeh N, Shahbani Zahiri H, Shariati P, Hajhosseini R - Avicenna J Med Biotechnol (2015 Jul-Sep)

No significant changes were observed in growth associated with ispA over-expression. Error bars indicate the standard error of the mean of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4508334&req=5

Figure 8: No significant changes were observed in growth associated with ispA over-expression. Error bars indicate the standard error of the mean of three independent experiments.
Mentions: In addition, it was observed that the reduction of CoQ10production in both E. coli BaTi and E. coli BrTi occurred simultaneously with an increase in CoQ8production when compared to E. coli Ba and E. coli Br (from 360 to 890 μg/g DCW in E. coli BaTi and 246 to 580 μg/g DCW in E. coli BrTi). However, induction by IPTG (0.05 mM) decreased the CoQ8biosynthesis as well in both E. coli BaTi (to 750 μg/g DCW) and E. coli BrTi (to 310 μg/g DCW) as shown in figures 6 and 7. The growth of recombinant strains was also studied and showed no significance changes by ispA over-expression (Figure 8).

Bottom Line: Over-expression of ispA under the control of stronger trc promoter, however, led to a severe decrease in CoQ 10 production in both recombinant E. coli Ba and E. coli Br strains, as reflected by reductions from 629±40 to 30±13 and 564±28 to 80±14 μg/g Dried Cell Weight (DCW), respectively.The results showed high level of FPP reduces endogenous CoQ 8 production as well and that CoQs are produced in a complimentary manner, as the increase in production of one decreases the production of the other.The reduction in CoQ 10 production can be a result of Dds inhibition by high FPP concentration.

View Article: PubMed Central - PubMed

Affiliation: Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.

ABSTRACT

Background: Coenzyme Q 10 (CoQ 10 ) is an isoprenoid component used widely in nutraceutical industries. Farnesyl diphosphate synthase (FPPS) is a responsible enzyme for biosynthesis of farnesyl diphosphate (FPP), a key precursor for CoQs production. This research involved investigating the effect of FPPS over-expression on CoQs production in engineered CoQ 10 -producing Escherichia coli (E. coli).

Methods: Two CoQ 10 -producing strains, as referred to E. coli Ba and E. coli Br, were transformed by the encoding gene for FPPS (ispA) under the control of either the trc or P BAD promoters.

Results: Over-expression of ispA under the control of P BAD promoter led to a relative increase in CoQ 10 production only in recombinant E. coli Br although induction by arabinose resulted in partial reduction of CoQ 10 production in both recombinant E. coli Ba and E. coli Br strains. Over-expression of ispA under the control of stronger trc promoter, however, led to a severe decrease in CoQ 10 production in both recombinant E. coli Ba and E. coli Br strains, as reflected by reductions from 629±40 to 30±13 and 564±28 to 80±14 μg/g Dried Cell Weight (DCW), respectively. The results showed high level of FPP reduces endogenous CoQ 8 production as well and that CoQs are produced in a complimentary manner, as the increase in production of one decreases the production of the other.

Conclusion: The reduction in CoQ 10 production can be a result of Dds inhibition by high FPP concentration. Therefore, more effort is needed to verify the role of intermediate metabolite concentration and to optimize production of CoQ 10 .

No MeSH data available.


Related in: MedlinePlus