Improved Stability and Half‐Life of Fluorinated Phosphotriesterase Using Rosetta

Yang, Ching-Yao; Renfrew, P. Douglas; Olsen, Anne O.; Zhang, Michelle; Yuvienco, Carlo; Bonneau, Richard; Montclare, Jin Kim

doi:10.1002/cbic.201402062

Cited by 21 publications

(18 citation statements)

References 41 publications

(20 reference statements)

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“…At the optimum pH range of 8.0-9.5 the Co 2+ OPT complex maintains thermostability < 45 o C, above which, the stability rapidly declines until deactivation at 60 o C 38 . Efforts to improve stability and function have focused on engineering OPT via rational design [39][40][41][42] , directed evolution 29 and the incorporation of non-canonical amino acids 37,43 .…”

Section: Introductionmentioning

confidence: 99%

Pollen-mimicking, enzyme-loaded microparticles to reduce organophosphate toxicity in managed pollinators

Chen¹,

Webb²,

Shariati³

et al. 2020

Preprint

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Pollinators support the production of 87 of the leading food crops worldwide and contribute over $15 billion to U.S. farm income. Organophosphates are a heavily used group of insecticides that pollinators can be exposed to, especially during crop pollination. Exposure to lethal or sub-lethal doses can impair fitness of wild and managed bees, risking pollination quality and food security. Here, we report a low-cost, scalable in vivo detoxification strategy for organophosphate insecticides involving encapsulation of phosphotriesterase (OPT) in pollen-mimicking microparticles (PMMs). We developed uniform and consumable PMMs capable of loading OPT at 90% efficiency and protecting OPT from degradation in the pH of a bee gut. Microcolonies of bumble bees (Bombus impatiens) fed malathion-contaminated pollen patties demonstrated 100% survival when fed OPT-PMMs but 0% survival with OPT alone and 0% survival with plain sucrose within 5 and 4 days, respectively. Thus, the detrimental effects of malathion were eliminated when bees consumed OPT-PMMs. This design presents a versatile and scalable treatment for managed pollinators to reduce risk from organophosphate insecticides, which can be integrated into supplemental feeds such as pollen patties or dietary syrup.

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Section: Introductionmentioning

confidence: 99%

Pollen-mimicking, enzyme-loaded microparticles to reduce organophosphate toxicity in managed pollinators

Chen¹,

Webb²,

Shariati³

et al. 2020

Preprint

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“…ncAAs containing long side-chain thiols were incorporated into β-lactamase, forming unnatural disulfide bond and exhibiting significantly enhanced thermostability both in vitro and in vivo [ 96 ]. Genetically incorporation of p ‐fluorophenylalanine (pFF) [ 97 ] or para ‐isothiocyanate phenylalanine (pNCSF) [ 98 ] was also capable of enhancing protein stability at elevated temperature.…”

Section: Increasing Therapeutic Agent Serum Half-livesmentioning

confidence: 99%

Therapeutic applications of genetic code expansion

Huang

Liu

2018

Synthetic and Systems Biotechnology

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In nature, a limited, conservative set of amino acids are utilized to synthesize proteins. Genetic code expansion technique reassigns codons and incorporates noncanonical amino acids (ncAAs) through orthogonal aminoacyl-tRNA synthetase (aaRS)/tRNA pairs. The past decade has witnessed the rapid growth in diversity and scope for therapeutic applications of this technology. Here, we provided an update on the recent progress using genetic code expansion in the following areas: antibody-drug conjugates (ADCs), bispecific antibodies (BsAb), immunotherapies, long-lasting protein therapeutics, biosynthesized peptides, engineered viruses and cells, as well as other therapeutic related applications, where the technique was used to elucidate the mechanisms of biotherapeutics and drug targets.

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“…43 The mutation Y309A was also found to improve expression (Table 3). 46 Introducing additional chemical functionalities, such as disulfide bridges 47 or fluorines, 45,48,49 provides further stability to PTE (see below). Computational design Recently, computational modeling has proven to be a powerful tool in protein engineering.…”

Section: Genetic Engineering Of Pte Outside the Catalytic Domainmentioning

confidence: 99%

Enhancing organophosphate hydrolase efficacy via protein engineering and immobilization strategies

Katyal

Chu

Montclare

2020

Annals of the New York Academy of Sciences

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Organophosphorus compounds (OPs), developed as pesticides and chemical warfare agents, are extremely toxic chemicals that pose a public health risk. Of the different detoxification strategies, organophosphate-hydrolyzing enzymes have attracted much attention, providing a potential route for detoxifying those exposed to OPs. Phosphotriesterase (PTE), also known as organophosphate hydrolase (OPH), is one such enzyme that has been extensively studied as a catalytic bioscavenger. In this review, we will discuss the protein engineering of PTE aimed toward improving the activity and stability of the enzyme. In order to make enzyme utilization in OP detoxification more favorable, enzyme immobilization provides an effective means to increase enzyme activity and stability. Here, we present several such strategies that enhance the storage and operational stability of PTE/OPH.

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Improved Stability and Half‐Life of Fluorinated Phosphotriesterase Using Rosetta

Cited by 21 publications

References 41 publications

Pollen-mimicking, enzyme-loaded microparticles to reduce organophosphate toxicity in managed pollinators

Pollen-mimicking, enzyme-loaded microparticles to reduce organophosphate toxicity in managed pollinators

Therapeutic applications of genetic code expansion

Enhancing organophosphate hydrolase efficacy via protein engineering and immobilization strategies

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