A Modified Method for Determination of Hippuric Acid in Urine by HPLC : İdrarda Hi̇ppuri̇k Asi̇t Tayi̇ni̇nde Kullanilabi̇lecek Bi̇r Modi̇fi̇ye HPLC Yöntemi̇

Duydu, Yalçın; Süzen, Sinan; Vural, Nevın; Erdem, Nurten; Uysal, Handan

doi:10.1501/eczfak_0000000330

Cited by 1 publication

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References 19 publications

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“…The column used was SB-C18 (4.6 mm×150 mm, Agilent, USA). The effluent was monitored at 225 nm and the total assay was carried out at 25°C (Duydu et al, 1999). Hippuric acid was purchased from Sigma.…”

Section: Determination Of Ha Levels In Urine By Hplcmentioning

confidence: 99%

Expression of phenylalanine ammonia lyase as an intracellularly free and extracellularly cell surface-immobilized enzyme on a gut microbe as a live biotherapeutic for phenylketonuria

Sun

Qian

Dong

et al. 2022

Sci. China Life Sci.

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Phenylketonuria (PKU), a disease resulting in the disability to degrade phenylalanine (Phe) is an inborn error with a 1 in 10,000 morbidity rate on average around the world which leads to neurotoxicity. As an potential alternative to a protein-restricted diet, oral intake of engineered probiotics degrading Phe inside the body is a promising treatment, currently at clinical stage II (Isabella, et al., 2018). However, limited transmembrane transport of Phe is a bottleneck to further improvement of the probiotic’s activity. Here, we achieved simultaneous degradation of Phe both intracellularly and extracellularly by expressing genes encoding the Phe-metabolizing enzyme phenylalanine ammonia lyase (PAL) as an intracellularly free and a cell surface-immobilized enzyme in Escherichia coli Nissle 1917 (EcN) which overcomes the transportation problem. The metabolic engineering strategy was also combined with strengthening of Phe transportation, transportation of PAL-catalyzed trans -cinnamic acid and fixation of released ammonia. Administration of our final synthetic strain TYS8500 with PAL both displayed on the cell surface and expressed inside the cell to the Pah F263S PKU mouse model reduced blood Phe concentration by 44.4% compared to the control EcN, independent of dietary protein intake. TYS8500 shows great potential in future applications for PKU therapy. Supporting Information The supporting information is available online at 10.1007/s11427-021-2137-3. The supporting materials are published as submitted, without typesetting or editing. The responsibility for scientific accuracy and content remains entirely with the authors.

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Section: Determination Of Ha Levels In Urine By Hplcmentioning

confidence: 99%

Expression of phenylalanine ammonia lyase as an intracellularly free and extracellularly cell surface-immobilized enzyme on a gut microbe as a live biotherapeutic for phenylketonuria

Sun

Qian

Dong

et al. 2022

Sci. China Life Sci.

View full text Add to dashboard Cite

show abstract

A Modified Method for Determination of Hippuric Acid in Urine by HPLC : İdrarda Hi̇ppuri̇k Asi̇t Tayi̇ni̇nde Kullanilabi̇lecek Bi̇r Modi̇fi̇ye HPLC Yöntemi̇

Abstract: In the present study, a fast and simple HPLC method has been described to determine urinary hippuric acid (HA)

Cited by 1 publication

References 19 publications

Expression of phenylalanine ammonia lyase as an intracellularly free and extracellularly cell surface-immobilized enzyme on a gut microbe as a live biotherapeutic for phenylketonuria

Expression of phenylalanine ammonia lyase as an intracellularly free and extracellularly cell surface-immobilized enzyme on a gut microbe as a live biotherapeutic for phenylketonuria

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