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Toxic product tolerating cell immobilized method and production process of 1,5-diaminopentane by immobilized cell

A technology for immobilizing cells and cells, applied in the field of microorganisms, can solve the problems of the large impact on the mechanical strength of the immobilized pellets and the inability to meet industrial production, and achieve the effects of easy scale-up of production, product clarification, and less equipment.

Inactive Publication Date: 2016-09-21
NANJING UNIV OF TECH
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, the ratio of the two and the corresponding gelation method have a great influence on the activity of immobilized cells and the mechanical strength of immobilized beads, especially in the decarboxylation similar to the conversion of L-lysine to 1,5-pentanediamine. During the reaction process, a large amount of carbon dioxide released has a more significant impact on the mechanical strength of the immobilized pellets, and the existing methods cannot meet the needs of industrial production

Method used

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  • Toxic product tolerating cell immobilized method and production process of 1,5-diaminopentane by immobilized cell
  • Toxic product tolerating cell immobilized method and production process of 1,5-diaminopentane by immobilized cell
  • Toxic product tolerating cell immobilized method and production process of 1,5-diaminopentane by immobilized cell

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Mixtures of different concentrations of PVA and 2% sodium alginate were used as immobilized materials, and immobilized cells were prepared by boric acid cross-linking method and freezing method, respectively. The specific operation method is: add polyvinyl alcohol into cold water, heat to 98°C to dissolve, add sodium alginate, heat to dissolve in a sterilizer, stir and cool to 40-50°C, add bacterial suspension to a final concentration of 10g (湿菌重) / L, stir well. Add the above mixed liquid into the cross-linking agent solution dropwise with a syringe or a peristaltic pump to obtain small balls with a diameter of 0.3-0.5 cm. The cross-linking agent of the boric acid cross-linking method uses a saturated boric acid solution containing 3% (mass percentage) calcium chloride; the freezing method cross-linking agent is a 3% (mass percentage) calcium chloride solution. For the pellets prepared by the boric acid cross-linking method, after standing at room temperature for 8 hou...

Embodiment 2

[0039] Immobilized cells were prepared by the freezing method in Example 1, wherein the mass fraction of sodium alginate was 2.0%, the mass fraction of polyvinyl alcohol was 2.0%, and the mass-volume ratio (g / 100mL) of the bacterial cell to the gel solution was 1-5. The relative activity of the immobilized pellets, except for the amount of bacteria in the transformation conditions, the other conditions are the same as in Example 1. The result is as figure 2 As shown, the concentration of 1,5-pentanediamine in the transformation solution increases with the increase of bacterial load, but the yield of 1,5-pentanediamine per unit mass of bacteria does not increase significantly when the bacterial load is greater than 3%. , and when the bacterial load was 3%, the 1,5-pentanediamine yield per unit mass of the immobilized cells was 4.41±0.09g / g (湿菌重) / h, compared with the control (4.89±0.46g / g (湿菌重) / h) compared with no significant difference, indicating that 3% of the bacteria l...

Embodiment 3

[0041] The immobilized cells were prepared by the freezing method in Example 1, and the mass fraction of the immobilized carrier sodium alginate (SA), polyvinyl alcohol (PVA), and the forming agent were tested by the central composite design (CCD) test in the response surface method analysis method. Factors such as mass fraction of calcium chloride solution (CaCl2), calcification time and freezing time are optimized. The levels of each factor are shown in Table 1, a total of 32 groups of tests, the conversion reaction conditions are the same as in Example 1. The test results are shown in Table 2. The test results were modeled and analyzed to obtain the regression equation:

[0042] Log10 (loudness vitality) = +1.38-1.81*SA+2.21E-003*PVA-0.26*CaCl 2 +0.039*calcification time-0.028*freezing time+0.074*SA*PVA+0.32*SA*CaCl 2 +0.053*SA*freezing time+0.27*SA 2 -0.014*PVA 2 -0.036*CaCl 2 2 -1.64E-003*calcification time 2 -1.94E-003*freezing time 2 -0.01*SA 2 *PVA-0.046*SA ...

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Abstract

The invention particularly relates a toxic product tolerating cell immobilized method and a continuous production process of 1,5-diaminopentane by immobilized cell through transformation. According to the invention, the ratio of polyvinyl alcohol to sodium alginate and a corresponding gel forming method are developed and optimized, the prepared immobilized cell has the advantages that damage to bacterial cells from 1,5-diaminopentane with high concentration is tolerated, the expansion character is better, the influence of release of a large amount of carbon dioxide gas on a structure in the transformation process is tolerated, the mechanical strength is high, and the service life is long; the activity of the prepared immobilized cell can be kept 98 percent or above after being used for 6 times in batches, the activity of the prepared immobilized cell can be kept 70 percent or above after being continuously used for 12 hours. The continuous production process provided by the invention has the advantages that needed devices are less, the operation is convenient, amplification of production scale is easy, and the degree of automation is high; no neutralizer is added in the production process, and the product is easy to clarify, separate and purify.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a method for immobilizing cells resistant to toxic products and a process for producing 1,5-pentanediamine by immobilized cells. Background technique [0002] 1,5-Pentanediamine (pentamethylenediamine for short), that is, cadaverine, is a nitrogenous base with biological activity widely present in organisms, which is produced when lysine undergoes decarboxylation reaction under the action of decarboxylase when protein decays product. In agricultural production, 1,5-pentanediamine can be used as a plant growth regulator to regulate plant aging process, stimulate plant growth, and promote fruit development; in addition, 1,5-pentanediamine can also improve the cold tolerance of plants; in In medicine, 1,5-pentanediamine is the precursor of deferoxamine B, which is clinically used to treat iron poisoning; and the most valuable application of 1,5-pentanediamine is as a nylon m...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/08C12P13/00
CPCC12N11/08C12N11/10C12P13/001
Inventor 陈可泉马伟超沈金山曹逊何育美欧阳平凯
Owner NANJING UNIV OF TECH
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