Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of engineering enterococcus faecalis in drugs for treating phenylketonuria

A technology for Enterococcus faecalis and Enterococcus, which is applied in the application, drug combination, genetic engineering and other directions, can solve the problems of reducing phenylalanine, taking a long time, and being expensive, and achieves the reduction of phenylalanine content, prevention// Treatment of phenylketonuria, the effect of treatment of phenylketonuria

Pending Publication Date: 2021-12-07
INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the treatment methods are to reduce the content of phenylalanine in breast milk or infant formula, and there is no specific drug or other curative therapy in clinical practice
Moreover, the existing methods require strict restriction on the intake of natural protein, are expensive, take a long time, and can only relieve symptoms but not cure them. The brain function development problems of young patients cannot be solved, and the quality of life of adult patients is difficult to guarantee.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of engineering enterococcus faecalis in drugs for treating phenylketonuria
  • Application of engineering enterococcus faecalis in drugs for treating phenylketonuria
  • Application of engineering enterococcus faecalis in drugs for treating phenylketonuria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] EXAMPLE 1. Recombinant human phenylalanine hydroxylase embodiment constructed plasmid vector

[0033] 1, experimental animals, instruments and reagents drugs

[0034] Restriction endonucleases KpnI and SacI and Gibson assembly premix night purchased from New England Biolabs company, the nucleic acid sequence of the synthetic technology services provided by Sangon Biotech (Shanghai) Co., lactic acid bacteria secreting plasmid pNZ8148 purchased from Hangzhou purple Yun Biological Technology Co., Ltd.

[0035] 2. Experimental Design

[0036] Acquiring human phenylalanine hydroxylase gene sequences from the NCBI database, using the reference software JCAT Enterococcus faecalis (ATCC47077) codon preference codon optimized gene sequence to give SEQ ID NO: 1 ( figure 1Indicates the change in CAI values ​​during the codon optimization, and the specific DNA sequence information is described below. Information comes from Reference 1, this sequence is obtained by synthesis in this exp...

Embodiment 2

[0045] Example 2. Screening of recombinant plasmids for the transformation of the host bacteria and the screening of the engineerus (EPR1901)

[0046] 1. Experimental animals, instruments and drug reagents

[0047] The recombinant plasmid P8NIS-PAH was obtained from the above experiment, and the MRS medium was purchased from Beijing Solabao Technology Co., Ltd., is available from Beijing Solabao Technology Co., Ltd. Sensitive cells were prepared by this laboratory.

[0048] 2. Experimental instrument and analysis method

[0049] Micropulser Electric Converter and Electric Tower Buy from US Bio-Rad.

[0050] 3. Experimental design

[0051] We will submit the above experiments, P8Nis-PAH, for the electrical conversion of the target host bacteria (Swyncus). The conversion method is as follows: The electric rotor is placed on ice, takes 100 μl of prepared sensitized cells, and plasmid DNA is added to 100 μL of sensitive state cells, ice bath 5 min; add plasmid and cell mixture to a pr...

Embodiment 3

[0054] Example 3. Engineering Swyncus (EPR1901) strain activation and vaccination fermentation, phenylalanine hydroxylase protein expression, concentration experiment and inducer NISIN concentration screening experiment

[0055] 1. Experimental strains, instruments and reagents

[0056] Cormenate peptide (NISIN) is purchased from Beijing Solabao Technology Co., Ltd., SDS-PAGE kit is purchased from Beijing Solabao Technology Co., Ltd., Kakamas Bright Blue Dyed Liquid from Beijing Solabao Technology Co., Ltd.

[0057] 2. Experimental instrument and analysis method

[0058] Electrophoretic tanks are purchased from Bio-Rad.

[0059] 3. Experimental design

[0060] Nisin-Controlled Expression (Nice) When the cell is like a bacteric peptide (Nisin), Nisk having a sensitive protein can identify milk pspococcus, so that NisR phosphorylation activates NISA promoters, resulting in Downstream gene expression. It is widely presented in Gram-positive bacteria that the product expressed in the ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of biology, and provides an application of engineering enterococcus faecalis capable of expressing human phenylalanine hydroxylase in drugs for treating phenylketonuria, and plasmids containing the human-derived phenylalanine hydroxylase gene optimized according to enterococcus faecalis codons are introduced into enterococcus faecalis, so that the human phenylalanine hydroxylase is expressed in the intestinal tract. Experimental results show that: the human phenylalanine hydroxylase is successfully expressed in the enterococcus faecalis through a gene engineering technology; the engineering enterococcus faecalis can obviously metabolize phenylalanine in an in-vitro culture medium system; meanwhile, the phenylketonuria can be remarkably treated by orally taking the engineering enterococcus faecalis, and the phenylalanine in the plasma of a suckling mouse with phenylketonuria can be reduced by 46.8% after the engineering enterococcus faecalis is used for treatment for 4 weeks. It is shown that the engineering enterococcus faecalis has an outstanding treatment effect in prevention and / or treatment of phenylketonuria.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, and more particularly relates to and / or application of an engineering faecalis prophylaxis phenylketonuria medicament. Background technique [0002] Phenylketonuria (of PKU) is an autosomal negative congenital hereditary metabolic disorders, mainly due to defects in the body such that the phenylalanine hydroxylase phenyl alanine phenylalanine metabolic pathway can not change generated tyrosyl acid, phenylalanine and phenylpyruvic acid such large accumulation occurs in vivo. The incidence of PKU is an average of 8.84 / 10 5 The main symptoms at 3-6 months of the child, 1 year old symptoms. The main clinical features of children with mental retardation as, depression, hyperactivity, autistic tendencies, eczema / skin scratch levy and EEG abnormalities. If you can get early diagnosis and early intervention, then the clinical manifestations do not occur in children of normal intelligence levels, abn...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/74C12N15/53A61K35/744A61P3/00
CPCC12N15/746C12N9/0071A61K35/744A61P3/00C12Y114/16001C12N2800/22
Inventor 王琰彭冉蒋建东马殊荣潘利斌
Owner INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com