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Application of Cellulomonadaceae as industrial biocatalyst

A Cellulomonas, biocatalyst technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as inability to meet industrialization requirements, limitations of enrichment and separation catalytic efficiency, etc.

Inactive Publication Date: 2016-11-23
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Metabolic hydrolysis bacteria or enzymes screened from nature include Moraxella sp.13, Leifsonia shinshuensis14, Enterobacter sp., Streptomyces matensi, Arthrobacter nicotianae, Achromobacterpiecchaudii, and Pseudomonas plecoglossicida16, and Lentinala edodes17, among which the xylosidase from Lentinala edodes catalyzes The highest efficiency, the Km for 10-DAXP is 1.79±0.07mM, and the kcat is 1.92s-1, but this efficiency cannot meet the above-mentioned DAXP industrialization requirements. After the enzyme is recombinantly expressed as an intracellular enzyme, it is difficult to enrich and separate. lead to a further limitation of the catalytic efficiency
The above findings indicate that the selective and efficient hydrolysis of the C-7 xylosidic bond of 10-DAXP is extremely challenging and requires novel catalytic and stable structures to complete

Method used

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  • Application of Cellulomonadaceae as industrial biocatalyst
  • Application of Cellulomonadaceae as industrial biocatalyst

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Preparation of crude enzyme solution

[0027] A medium (pH 7.0 before sterilization) containing 1% starch, 0.2% peptone, 0.2% yeast extract, and 0.2% dipotassium hydrogenphosphate was used as the medium. Cellulomonas flavigena bacteria of Cellulomonadaceae (Cellulomonadaceae) were inoculated into the above-mentioned medium, and cultured with shaking at 150 rpm at 30° C. for 24 hours. After the fermentation culture is finished, after the cells are removed by centrifugation, the supernatant is taken, which is the crude enzyme solution.

Embodiment 2

[0029] Preparation of crude enzyme solution

[0030] A medium (pH 7.0 before sterilization) containing 1% starch, 0.2% peptone, 0.2% yeast extract, and 0.2% dipotassium hydrogenphosphate was used as the medium. Oerskovia enterophila or Oerskovia turbata bacteria of Cellulomonadaceae (Cellulomonadaceae) were inoculated into the above-mentioned culture medium, and cultured with shaking at 150 rpm at 30° C. for 24 hours. After the fermentation culture is finished, after the cells are removed by centrifugation, the supernatant is taken, which is the crude enzyme solution.

Embodiment 3

[0032] Preparation of crude enzyme solution

[0033] A medium (pH 7.0 before sterilization) containing 1% starch, 0.2% peptone, 0.2% yeast extract, and 0.2% dipotassium hydrogenphosphate was used as the medium. Demequina aestuarii bacteria of the family Cellulomonadaceae were inoculated into the above-mentioned culture medium, and cultured with shaking at 150 rpm at 30° C. for 24 hours. After the fermentation culture is finished, after the cells are removed by centrifugation, the supernatant is taken, which is the crude enzyme solution.

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Abstract

The invention provides application of Cellulomonadaceae as an industrial biocatalyst, belonging to the field of biochemical engineering. The Cellulomonadaceae provided by the invention is used for hydrolysis of xyloside and glucoside bonds of glucoside compounds so as to prepare corresponding aglycones or semi-hydrolysates. The Cellulomonadaceae is applicable to the field of biological medicine for preparation of drug intermediates and also applicable to the fields of food, agriculture and food for preparation of monosaccharides or corresponding aglycones. The invention provides application of the Cellulomonadaceae as the industrial biocatalyst; and good effect is obtained when the Cellulomonadaceae is applied to hydrolysis of a mixture of taxane xyloside and astragaloside extracted from plant materials.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and specifically relates to the application of Cellulomonasaceae as an industrial biocatalyst. Background technique [0002] Taxane drugs, representative drugs such as paclitaxel and docetaxel, are currently one of the most important anticancer drugs (Nature Reviews Cancer 4, 253-265 (2004)). However, due to the complex structure and the difficulty of chemical synthesis, it mainly comes from the production process based on yew plants (Current Medicinal Chemistry 20,880-891(2013)). The content of paclitaxel itself in the yew plants is not high (0.01% in bark of Taxus brevifolia), and there was once a serious resource crisis. However, there are a large number of taxane precursor compounds or derivatives in some yew plants. For example, the content of 10-DAXP in the branches and leaves of Taxus yunnanensis and Taxus mairei is at the same level as 10-DAB (~0.1%), so It is also an important poten...

Claims

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Application Information

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IPC IPC(8): C12P33/20C12P17/02C12R1/01
Inventor 杨凌窦同意栾宏伟葛广波王家悦
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI