Mevalonate kinase from the larva of the fleshfly, Sarcophaga bullata

Goodfellow, Robert D.; Barnes, Francis J.

doi:10.1016/0020-1790(71)90043-6

Cited by 20 publications

(3 citation statements)

References 32 publications

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“…Sarcophaga bullata MK has been partially characterized, showing low affinities for MVA and ATP. 18 MKs with higher tolerance for long-chain isoprenoids and higher MVA and ATP affinities have previously received little attention with regards to lycopene production. Directed evolution has emerged as a ubiquitous technique to enhance the stability, activity, and selectivity of enzymes.…”

Section: Introductionmentioning

confidence: 99%

Directed evolution of mevalonate kinase inEscherichia coliby random mutagenesis for improved lycopene

Chen

Liu

et al. 2018

RSC Adv.

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a Lycopene is a terpenoid pigment that has diverse applications in the fields of food and medicine. Metabolic engineering in microbial hosts has shown that mevalonate kinase (MK, EC2.7.1.366) is one of the ratelimiting enzymes in the lycopene synthetic pathway. In this study, a directed evolution strategy in Escherichia coli was used to optimize the activity of Saccharomyces cerevisiae MK. Using three rounds of error-prone PCR; screening the development of a lycopene-dependent color reaction; and combinatorial site-specific saturation mutagenesis, three activity-enhancing mutations were identified: V13D, S148I, and V301E. V13D was near the MK catalytic center, in the b-sheet that forms a salt-bridge with nearby Arg-248. S148I was located in the a-helix lid and improved the stability of the a-helix. V301E may increase MK folding by influencing its secondary structure. The K m (RS)-mevalonate of purified mutant MK decreased by 74% compared with the K m (RS)-mevalonate of the wild-type MK, and the K cat (RS)-mevalonate was improved by 26% compared with wild type. Fermentation experiments revealed that lycopene production of the mutant MK increased 2.4-fold compared with wild-type MK.

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Section: Introductionmentioning

confidence: 99%

Directed evolution of mevalonate kinase inEscherichia coliby random mutagenesis for improved lycopene

Chen

Liu

et al. 2018

RSC Adv.

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“…aegypti [ 19 – 21 ]; however, the catalytic properties of these enzymes have been poorly characterized. A MVK from Sarcophaga bullata was partially characterized, showing low affinities for MA and ATP [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

Negative Feedbacks by Isoprenoids on a Mevalonate Kinase Expressed in the Corpora Allata of Mosquitoes

2015

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BackgroundJuvenile hormones (JH) regulate development and reproductive maturation in insects. JHs are synthesized through the mevalonate pathway (MVAP), an ancient metabolic pathway present in the three domains of life. Mevalonate kinase (MVK) is a key enzyme in the MVAP. MVK catalyzes the synthesis of phosphomevalonate (PM) by transferring the γ-phosphoryl group from ATP to the C5 hydroxyl oxygen of mevalonic acid (MA). Despite the importance of MVKs, these enzymes have been poorly characterized in insects.ResultsWe functionally characterized an Aedes aegypti MVK (AaMVK) expressed in the corpora allata (CA) of the mosquito. AaMVK displayed its activity in the presence of metal cofactors. Different nucleotides were used by AaMVK as phosphoryl donors. In the presence of Mg2+, the enzyme has higher affinity for MA than ATP. The activity of AaMVK was regulated by feedback inhibition from long-chain isoprenoids, such as geranyl diphosphate (GPP) and farnesyl diphosphate (FPP).Conclusions AaMVK exhibited efficient inhibition by GPP and FPP (Ki less than 1 μM), and none by isopentenyl pyrophosphate (IPP) and dimethyl allyl pyrophosphate (DPPM). These results suggest that GPP and FPP might act as physiological inhibitors in the synthesis of isoprenoids in the CA of mosquitoes. Changing MVK activity can alter the flux of precursors and therefore regulate juvenile hormone biosynthesis.

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“…The enzyme responsible for the first step in the biosynthesis of farnesyl pyrophosphate (2) from 6, mevalonic kinase, has been obtained from a number of organisms,92-96 including the fly, Sarcophaga bullata. 97 Further work with S. bullata98 demonstrated that along with trans, trans-tarnesoi (1) and transnerolidol (11), geranylgeraniol (16) was the most complex metabolite labeled by [2-14C]mevalonate (6). Mevalonic inhibited in vitro by both geranyl pyrophosphate (10) and farnesyl pyrophosphate (2).…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis of sesquiterpenes

Cordell¹

1976

Chem. Rev.

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Mevalonate kinase from the larva of the fleshfly, Sarcophaga bullata

Cited by 20 publications

References 32 publications

Directed evolution of mevalonate kinase inEscherichia coliby random mutagenesis for improved lycopene

Directed evolution of mevalonate kinase inEscherichia coliby random mutagenesis for improved lycopene

Negative Feedbacks by Isoprenoids on a Mevalonate Kinase Expressed in the Corpora Allata of Mosquitoes

Biosynthesis of sesquiterpenes

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