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Immobilization reaction device and reaction method using immobilization technology

A reaction device and reaction technology, applied in the field of bioengineering, can solve the problems of not providing reaction devices and methods, and achieve the effect of improving contact

Inactive Publication Date: 2016-10-19
GENEHARBOR HONG KONG BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this patent does not provide a reaction device and method for industrial production using this type of immobilized enzyme or immobilized cells

Method used

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  • Immobilization reaction device and reaction method using immobilization technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Embodiment 1: the production of creatine phosphate

[0081] Design PCR primers according to the sequence of Chinese Patent Application Publication No. CN102808006, specifically:

[0082] Upstream primer CPK1:

[0083] 5'-GGGAATTCCATATGCCGTTCGGCAACACCCACAAC-3'

[0084] Downstream primer CPK2:

[0085] 5'-CCGCTCGAGCTTCTGCGCGGGGATCATGTCGTCG-3'

[0086] According to the method described in Chinese Patent Application Publication No. CN102808006, the creatine kinase gene was obtained and connected to pET-21a to obtain pET-21a(+)-CPK, which was transformed into Escherichia coli BL21(DE3).

[0087] Inoculate above-mentioned bacterial strain in 4ml LB medium (containing 100ug / ml ampicillin), cultivate 16 hours as primary seed liquid, inoculate in 100ml LB medium (containing 100ug / ml ampicillin) by 1% quantity ratio after completion . The secondary seeds were cultivated at 37° C. and 200 rpm for 10 hours, and then 1% of the secondary seeds were inoculated into a 100 L fermen...

Embodiment 2

[0093] Example 2: Production of S-adenosylmethionine

[0094] Design PCR primers according to the sequence of Chinese patent CN101134948B, specifically:

[0095] Upstream primer parental S1:

[0096] 5'-AGCCTAGGTTAATTAACTTTAAGAAGGAGATACATATGAGAAACATAATTGTAAA-3'

[0097] Downstream primer parent S2:

[0098] 5'-ATAAGCTCAGCGGCGCGCCTTAGAATGTAGTTACTTTTTCCTTCA-3'

[0099] According to the Chinese patent CN101134948B, M.Jannaschii ATCC43067 (ATCC, USA) DNA was used as a substrate for PCR, and the PCR product was treated with restriction endonucleases PacI and AscI and ligated into pGEMT-EASY (Promega, USA) , to obtain pGEMT-SAM. The recombinant expression vector was transformed into Escherichia coli HB101 to obtain a strain expressing S-adenosylmethionine synthetase.

[0100] The bacterial strain containing pGEMT-SAM was inoculated in 5ml LB medium (containing 50mg / L ampicillin) and cultivated for 16 hours as the primary seed liquid, and was inoculated to 100ml LB medium (conta...

Embodiment 3

[0107] According to the preparation method of Example 1, the creatine kinase gene and E. coli cells expressing the gene were obtained, and fermentation was carried out to prepare creatine kinase immobilized E. coli cells on the carrier. The shape of the carrier was a strip: 22 cm long and 5 cm wide , Thickness 5mm, real weight 45.8g.

[0108] The above prepared carrier bearing immobilized E. coli cells with creatine kinase was installed in an immobilized enzyme / immobilized cell reactor. The reactor is a cylinder made of organic glass with a height of 1.8 cm and a radius of 9 cm. Use a knife to trim about 3 cm from the head and tail of the above-mentioned carrier with a slope of 45°, and roll it into a homogeneous cylinder with a height of 1.5 cm and a radius of 9 cm, weighing 30 g. The cylinder was inserted into the reactor so that it was tight enough to meet the Class 3 standard described in Table 1 and that there was no gap between the side wall and the inner wall of the re...

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Abstract

The invention provides an immobilization reaction device and a reaction method using immobilization technology. The immobilization reaction device comprises a column reactor with an inlet and outlet; the reaction device has a top and a bottom facing each other, and an internal cavity with defined side walls and for connecting the top and the bottom.

Description

technical field [0001] The invention belongs to the field of bioengineering, in particular to an immobilized reaction device and a reaction method using immobilized technology. Background technique [0002] The use of enzymes to produce various chemicals is a common technology today, and its advantages lie in low energy consumption and green production. But the liquid enzyme method has its limitations: the liquid enzyme can only be used once. Therefore, enzymes often occupy a high proportion in the entire production cost, which is the main reason for the high cost of enzymatic production. In addition, the extensive use of liquid enzymes runs counter to the original intention of green industry, and liquid enzymes remain in the reaction solution after the enzymatic reaction, which increases the difficulty of purifying products. The use of immobilized enzymes and immobilized cells can avoid the above disadvantages. [0003] Chinese patent CN1982445B discloses a method and ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/40C12P1/00C12P1/04
CPCC12M21/18C12M25/18C12M29/10C12M41/40C12P13/001C12P19/40C12M1/00C12M1/40C12M25/00C12M29/00C12M41/26C12P1/00C12P1/04C12P13/00
Inventor 潘永强萧游龙陈军明沈冬王骏孙梁
Owner GENEHARBOR HONG KONG BIOTECH LTD
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