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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

Plasmid pDispA was introduced into E. coli Br, and the coenzyme Qs content was quantified in the resulting strain, as referred to E. coli BrDi. Numbers in brackets indicate arabinose mM concentration in cultures. Error bars indicate the standard error of the mean of three independent experiments.
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Figure 5: Plasmid pDispA was introduced into E. coli Br, and the coenzyme Qs content was quantified in the resulting strain, as referred to E. coli BrDi. Numbers in brackets indicate arabinose mM concentration in cultures. Error bars indicate the standard error of the mean of three independent experiments.

Mentions: In the absence of inducer, the expression of ispA under the control of PBADpromoter does not have significant effect on CoQ10production in E. coli BaDi, with negligible change from 629±40 to 636±35 μg/g DCW (Figure 4), while relative increase was observed in E. coli BrDi from 564±28 to 750±28 μg/g DCW (Figure 5). However, once induction by arabinose was applied, relative reduction in CoQ10production was obtained which was proportional to the concentration of arabinose in both strains. In the absence of induction, the CoQ8production demonstrated roughly a twofold reduction in E. coli BrDi (from 246±24 to 117±13 μg/g DCW), while no significant change (from 361±23 to 373±28 μg/g DCW) was observed in E. coli BaDi. Insignificant reduction in CoQ8production was observed in both strains with increased arabinose concentration (Figures 4 and 5). As a whole, the results showed that CoQs production in both ispA-expressing strains is inhibited once the induction is applied.


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)

Plasmid pDispA was introduced into E. coli Br, and the coenzyme Qs content was quantified in the resulting strain, as referred to E. coli BrDi. Numbers in brackets indicate arabinose mM concentration in cultures. 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 5: Plasmid pDispA was introduced into E. coli Br, and the coenzyme Qs content was quantified in the resulting strain, as referred to E. coli BrDi. Numbers in brackets indicate arabinose mM concentration in cultures. Error bars indicate the standard error of the mean of three independent experiments.
Mentions: In the absence of inducer, the expression of ispA under the control of PBADpromoter does not have significant effect on CoQ10production in E. coli BaDi, with negligible change from 629±40 to 636±35 μg/g DCW (Figure 4), while relative increase was observed in E. coli BrDi from 564±28 to 750±28 μg/g DCW (Figure 5). However, once induction by arabinose was applied, relative reduction in CoQ10production was obtained which was proportional to the concentration of arabinose in both strains. In the absence of induction, the CoQ8production demonstrated roughly a twofold reduction in E. coli BrDi (from 246±24 to 117±13 μg/g DCW), while no significant change (from 361±23 to 373±28 μg/g DCW) was observed in E. coli BaDi. Insignificant reduction in CoQ8production was observed in both strains with increased arabinose concentration (Figures 4 and 5). As a whole, the results showed that CoQs production in both ispA-expressing strains is inhibited once the induction is applied.

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