摘要:
A GMP adapted to provide the PAL gene for the treatment of PKU when administered orally. The GMP of the present invention may include a probiotic, a PAL gene to be expressed using the probiotic, wherein the PAL gene is functionally attached to a promoter and a ribosome binding site, and may be codon-optimized for expression in a certain host organism. A method of treating the metabolic disease of PKU by oral administration and ingestion of a GMP is also provided.
摘要:
The subject technology generally relates to biosynthesis of styrene. Certain embodiments of the subject technology is based, in part, on the recognition that phenylalanine can be converted to styrene by a two-step pathway of deamination and decarboxylation, with trans-cinnamic acid (tCA) as the intermediate. Two types of enzymes are directly involved in this process, phenylalanine ammonia lyase (PAL), which converts phenylalanine to tCA, and cinnamic acid decarboxylase, which coverts tCA to styrene. Host cells expressing these two types of enzymes can be cultured in bioreactor to produce styrene from renewable substrates such as glucose.
摘要:
The subject technology generally relates to biosynthesis of styrene. Certain embodiments of the subject technology is based, in part, on the recognition that phenylalanine can be converted to styrene by a two-step pathway of deamination and de-carboxylation, with trans-cinnamic acid (tCA) as the intermediate. Two types of enzymes are directly involved in this process, phenylalanine ammonia lyase (PAL), which converts phenylalanine to tCA, and cinnamic acid decarboxylase, which coverts tCA to styrene. Host cells expressing these two types of enzymes can be cultured in bioreactor to produce styrene from renewable substrates such as glucose.
摘要:
The present invention provides a method for purifying an Anabaena variabilis phenylalanine ammonia-lyase (AvPAL) variant with minimal aggregation. The method comprises the steps of (a) lysing bacterial cells containing the AvPAL variant by homogenization to generate a cell lysate; (b) heating the cell lysate to 65°C for 30 to 120 minutes; (c) centrifuging the heated cell lysate, wherein a supernatant comprising the AvPAL variant is retained; (d) filtering the supernatant to remove precipitates; and (e) separating the AvPAL variant from contaminating proteins by sequential chromatography over an anion exchange (AIEX) column followed by a hydrophobic interaction (HIC) column, wherein the eluate from the HIC column comprises the AvPAL variant, wherein the cysteine residues at positions 503 and 565 of said AvPAL variant have been substituted by serine residues (SEQ ID NO:11).
摘要:
A GMP adapted to provide the PAL gene for the treatment of PKU when administered orally. The GMP of the present invention may include a probiotic, a PAL gene to be expressed using the probiotic, wherein the PAL gene is functionally attached to a promoter and a ribosome binding site, and may be codon-optimized for expression in a certain host organism. A method of treating the metabolic disease of PKU by oral administration and ingestion of a GMP is also provided.
摘要:
It is disclosed a non-naturally occurring microbial organism comprising at least one exogenous nucleic acid encoding an enzyme, or a portion thereof, selected from the group of ammonia lyase. Preferably, the enzyme is PAL3, and the at least one exogenous nucleic acid is obtained from Arabidopsis thaliana . The non-naturally occurring microbial organism has an increased resistance to biochemical stress compared to the starting microbial organism, as induced for instance by oxidative stress or organic acid stress. Preferably, the non-naturally occurring microbial organism is a yeast and it may be used for fermenting a carbon source obtained from a ligno-cellulosic feedstock.
摘要:
Provided herein are methods for treating a subject suffering from phenylketonuria by administering a phenylalanine hydroxylase ('PAH') and/or a phenylalanine ammonia lyase ('PAL') to the subject under conditions effective to deliver the phenylalanine 4-hydroxylase and/or PAL to the subject's small intestine. Also provided are methods for increasing the therapeutic activity of a phenylalanine 4-hydroxylase by thiolating the phenylalanine 4-hydroxylase. In addition, provided are oral dosage forms that include a phenylalanine 4-hydroxylase and/or a PAL and an enteric coating.