8 results about "Resting Cell" patented technology

The invention discloses a vidarabine monophosphate biosynthesis method based on an ATP (adenosine triphosphate) regeneration system, and belongs to the technical field of biomedical engineering. According to the method, riboside arabinoside and ATP serve as reaction substrates, the target product riboside arabinoside monophosphate is synthesized through double-enzyme catalysis, double enzymes are deoxynucleoside kinase and polyphosphate kinase, the deoxynucleoside kinase is mainly used for catalyzing riboside arabinoside to synthesize riboside arabinoside monophosphate, and the polyphosphate kinase is used for catalyzing riboside arabinoside to synthesize riboside arabinoside monophosphate. Polyphosphokinase is mainly used for catalyzing ADP (adenosine diphosphate) and polyphosphoric acid or polyphosphate to regenerate to obtain ATP (adenosine triphosphate). The method can be used for a pure enzyme reaction system, a crude enzyme liquid reaction system or a resting cell catalysis system. The method disclosed by the invention is environment-friendly, realizes low-cost efficient conversion of a substrate through coupled ATP regeneration, plays a significant role in promoting biosynthesis of arabinoside monophosphate series nucleoside drugs, and has application potential and economic value.

The invention discloses a method for synthesizing 11 alpha hydroxyprogesterone by using cytochrome P450 enzyme recombinant bacteria, and belongs to the technical field of biology. The method comprises the following steps: firstly, constructing CYP219A1 gene and reduction chaperone Pdx / Pdr gene expression vectors from neosphingosinearomaphilus, and transferring the CYP219A1 gene and reduction chaperone Pdx / Pdr gene expression vectors into escherichia coli to obtain whole-cell transformed recombinant bacteria; and after culture, feeding a substrate progesterone by using a resting cell method for biotransformation to obtain a 11 alpha-hydroxyprogesterone product. According to the CYP219A1 mutant strain provided by the invention, the conversion rate of progesterone is as high as 99%, and the 11 alpha site selectivity is 74%.

The invention relates to the technical field of biological pharmacy, and discloses a mutant strain for efficient biotransformation synthesis of miglitol, a screening method and application. A compound mutation strategy of ARTP mutagenesis and NTG mutagenesis is utilized, a rapid screening method based on carbonyl color development is combined, a mutant strain capable of efficiently converting and synthesizing a miglittol key intermediate is obtained, the mutant strain is named as Gluconobacter oxydans G1036, and the enzyme activity of the mutant strain is 61.7 U / mL and is improved by about 48% compared with that of an original strain. A process for producing 6NSL by catalyzing NEHG through resting cells of the mutant strain is researched, 73.53 g / L of 6NSL is produced by catalyzing 80 g / L of a substrate within 48 h, and compared with 80.37% of an original strain, the 6NSL is improved by 19.63%. According to the present invention, the research on the process for repeatedly and catalytically producing the 6NSL by using the resting cells of the mutant strain in multiple batches shows the good catalytic performance, and 3 batches can be efficiently utilized to accumulatively produce 220.59 g / L of the 6NSL within 144 h;

This invention discloses a bio-preparation method for a key chiral intermediate of erlotinib ethyl ester. The method utilizes resting cells of *Saccharomyces cerevisiae* ZJPH1807 as a catalyst to prepare (R)-1-[4-chloro-2-(3-methyl-1H-pyrazol-1-yl)phenyl]-2,2,2-trifluoroethanol via biocatalysis of 1-[4-chloro-2-(3-methyl-1H-pyrazol-1-yl)phenyl]-2,2,2-trifluoroethanol. The product obtained using this strain exhibits high optical purity (ee value > 99.9%) and a reaction yield of 86.4%. The process is simple, environmentally friendly, and uses microbial cells as the biocatalyst, resulting in low cost.

The invention discloses fermented vegetable oil and a production method thereof. The invention belongs to a whole-cell enzymatic hydrolysis fermentation process of resting cells, which comprises the following steps: firstly, carrying out high-density fermentation to obtain a saccharomycetes liquid, then separating and washing to obtain clean high-activity saccharomycetes, and mixing the high-activity saccharomycetes, water and vegetable oil in proportion; vegetable oil is fermented by using resting yeast thalli through a whole-cell enzymolysis fermentation process under the conditions of specific temperature, oxygen supply and pH, and after fermentation is finished, secondary centrifugal filtration is performed to obtain the fermented vegetable oil. And compared with the fermented vegetable oil obtained by the conventional microbial fermentation process. The obtained fermented oil has the advantages of fewer impurities, lighter color, cleaner oil, low ash content, stronger oxidation stability, longer shelf life and the like, is rich in free fatty acids, flavonoids, polysaccharides and natural glycolipids compared with natural vegetable oil crude oil, has better skin moisturizing, repairing, soothing and anti-oxidation effects and the like, and is suitable for being used as a skin care product. Good economic values and application prospects are realized in the fields of beauty, skin care and washing.

This invention discloses a Valenciane oxidase mutant and its application in the preparation of naringin. The invention constructs a gene library of the HPO mutant using site-directed mutagenesis. In vitro catalytic experiments using resting cells show that, under the same conditions, the optimal HPO mutant (HPO_G302G-V480A-V482A-A484A, whose amino acid sequence is shown in SEQ ID No. 4 and nucleotide sequence in SEQ ID No. 5) exhibits a catalytic efficiency 2.54 times that of the wild type, thereby increasing substrate utilization efficiency and improving the yield of the target compound, naringin. Furthermore, the bio-preparation method for naringin described in this invention is simple to operate, environmentally friendly, and uses mild reaction conditions, showing great application potential. In addition, this invention is the first report of successfully constructing a beneficial HPO mutant for high-yield naringin production.

Disclosed herein are in vitro methods for detecting the presence of metabolically altered cells. Also disclosed herein are in vitro methods for detecting the presence of stationary cells. Disclosed herein are in vitro methods for detecting and predicting the presence of cancer. Disclosed herein are in vitro methods for monitoring response to anti-cancer therapy. The present disclosure analyzes the expression of at least one biomarker of pluripotent stem cells for detecting or predicting or monitoring cancer. Disclosed herein are related uses of at least one biomarker of pluripotent stem cell markers and methods of treatment including in vitro detection or prediction methods.

The invention discloses a 4-chlorobiphenyl degradation method based on a pseudomonas putida B6-2 resting cell microbial inoculum, and relates to the field of microbial degradation, the method comprises the following steps: mixing 4-chlorobiphenyl with the pseudomonas putida B6-2 resting cell microbial inoculum, and placing under an oscillation condition for reaction; the preparation method of the pseudomonas putida B6-2 resting cell microbial inoculum comprises the following steps: inoculating a strain pseudomonas putida B6-2 into an LB culture medium, and culturing for 12 hours in a shaking table at the temperature of 30 DEG C and the speed of 200 rpm to obtain a seed culture solution; centrifuging the seed culture solution for 10 minutes at the temperature of 4 DEG C and the speed of 4000 rpm, collecting thalli, resuspending the thalli by using a sterile MSM culture medium, repeating the steps of centrifuging and resuspending for three times, adding the MSM culture medium to adjust the OD600 of the system to be 15, and finally performing starvation treatment on the resuspended thalli to obtain the resting cell microbial inoculum. The resting cell microbial inoculum of the pseudomonas putida B6-2 can degrade more than 90% of 4-chlorobiphenyl within 35 hours, the preparation and application processes are simple, and an efficient, economical and environment-friendly effective measure is provided for degradation of the 4-chlorobiphenyl.