Carrier-free double-immobilized biocatalyst for displaying biological enzyme on surface as well as preparation method and application of carrier-free double-immobilized biocatalyst

Abstract

The invention discloses a carrier-free double-immobilized efficient biocatalyst for displaying biological enzyme on the surface as well as a preparation method and application of the carrier-free double-immobilized efficient biocatalyst, and belongs to the technical field of biochemical engineering and fermentation engineering. The enzyme is fixed on the surface of the host cell by utilizing a cell surface display technology, and cell clustering is realized by utilizing cell expression flocculation protein, so that the enzyme is fixed in a reactor, flocculation cell particles are settled by dead weight without centrifugal operation, product separation and continuous recycling of a catalyst can be realized, the production link is simplified, the production intensity is increased, and the production cost is reduced. The production cost is reduced; in addition, the immobilized cells are high in density, a cell population effect is formed, the environmental stress tolerance is further improved, the product concentration inhibition pressure is reduced, and the method is widely applied to large-scale green biological manufacturing of various high-added-value chemicals. The preparation method of the biocatalyst disclosed by the invention lays a good foundation for developing biocatalysts with completely independent intellectual property rights and green biological manufacturing of chemicals in China.

Description

technical field [0001] The invention belongs to the technical field of biochemical engineering and fermentation engineering, in particular to the development and optimization of a carrier-free double immobilization biocatalyst and its application in the process of biocatalytic transformation and product separation and purification. Background technique [0002] As the world's largest manufacturing country, the chemical industry is an important basic industry for my country's national economy and national defense industry. However, the current economic and social development faces many challenges, such as increasing energy demand, increasingly depleted land, water and fossil resources, and increasing global warming. Therefore, developing a green and sustainable manufacturing model is an inevitable trend in the development of my country's chemical industry. [0003] In recent years, the continuous progress of bio-manufacturing technology has provided green power for the trans...

AI Technical Summary

Problems solved by technology

Among them, enzyme preparations have high catalytic activity, but there are disadvantages such as many enzyme preparation steps, high difficulty, easy inactivation and difficult recycling, difficult separation and purification of downstream products, and high production costs.

In industrial production processes, enzyme immobilization is generally used to improve enzyme utilization, but traditional immobilization methods also have the problem of mass transfer resistance that affects catalytic efficiency (patents CN104480075A and CN105274174A)

The whole-cell catalyst containing biological enzymes effectively overcomes the disadvantages of using enzyme preparations, but the product is easily degraded by intracellular enzymes, the transmembrane resistance affects production efficiency, and the separation and purification process requires the use of large centrifugal equipment and huge energy consumption. Problems that cannot be ignored (patent CN201611254213.2)

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