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Biological material for enhancing electron transfer efficiency as well as preparation method and application of biological material

A biological material and electron transfer technology, applied in the biological field, can solve the problems of waste, pollution of environmental resources, etc., and achieve the effect of not being restricted by geographical areas, having a wide range of sources, and improving the sensitivity and efficiency of the reaction.

Pending Publication Date: 2022-04-29
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] A large amount of plant waste is produced every year on the earth, most of which are directly burned by farmers, which will not only pollute the environment but also cause a waste of resources

Method used

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  • Biological material for enhancing electron transfer efficiency as well as preparation method and application of biological material
  • Biological material for enhancing electron transfer efficiency as well as preparation method and application of biological material
  • Biological material for enhancing electron transfer efficiency as well as preparation method and application of biological material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1: Acquisition of Gene and Construction of Expression Vector

[0063] 1. Gene acquisition

[0064] The anchoring protein gene Lot is derived from Escherichia coli (Chang-ye Hui, Yan Guo, Lisa Liu, et al. Development of a novel bacterial surface display system using truncated OmpTas an anchoring motif. Biotechnol Lett. 2019:41:763–777) , whose nucleotide sequence is shown in SEQ ID NO.1, which was chemically synthesized by Suzhou Jinweizhi Company into the pUC-GW-Kan vector to obtain the pUC-Lot vector.

[0065] The anchor protein gene MipA is derived from Escherichia coli (Mee-Jung Han.Novel bacterial surfacedisplay system based on the Escherichia coli proteinMipA.J.Microbiol.Biotechnol. 2020.30(7):1097-1103), and its nucleotide sequence is as SEQ ID As shown in NO.2, the pUC-MipA vector was obtained by chemical synthesis on the pUC-GW-Kan vector by Suzhou Jinweizhi Company.

[0066] The anchor protein gene Spycather is derived from Streptococcus pyogenes (H.B...

Embodiment 2

[0161] Example 2: NADH standard curve drawing

[0162] Dilute NADH solutions of different concentrations, measure the absorption peak at 340 nm, and draw the NADH standard curve with the absorption peak value at 340 nm to prepare for the enzymatic activity calculation later.

[0163] A under different concentrations of NADH 340 The values ​​are as follows:

[0164] NADH concentration mM 0 0.05 0.1 0.15 0.2 0.25 A 340

[0165] NADH standard curve see Figure 10 .

Embodiment 3

[0166] Example 3: Enzymatic activity assay of surface-displayed whole-cell catalysts

[0167] All the above successfully constructed plasmids were transformed into E. coli BL21 (DE3) competent cells respectively to obtain expression strains, which were stored in a -80°C refrigerator.

[0168] The strains were transferred to 10 mL of LB liquid medium from a glycerol tube stored in a -80°C freezer and activated overnight. The next day, 1 mL of seed solution was transferred to 100 mL of LB liquid medium, and cultured on a shaker at 37°C. When its OD = 0.6, let it cool, add 50 μl IPTG, and incubate overnight at 16°C on a shaker. The next day, 6000rpm, 4°C, centrifuged for 5 min to collect the bacteria, washed twice with 50mM HEPES buffer (pH 7.0) to completely wash off the medium, and finally suspended the bacteria in an appropriate HEPES buffer to make the OD. 600 = 1.0.

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Abstract

The invention provides a biological material for enhancing electron transfer efficiency as well as a preparation method and application of the biological material. The biological material is prepared by mixing a recombinant strain containing a TtGH8 gene, a recombinant strain containing an SxA gene, a recombinant strain containing an FsUA gene, a recombinant strain containing an XylB gene and an XylC gene, and a recombinant strain containing an XylA gene and an XylD gene, and then mixing with a reaction substrate. Exogenous genes related to hemicellulose degradation and xylose degradation are expressed in escherichia coli, different anchoring protein genes are selected at the same time, hemicellulose and xylose degradation gene expression products are anchored to an extracellular membrane, and electron transfer can be achieved without a substrate entering bacteria. The degradation process of the biological material is accompanied by the generation of NADH and electrons, so that the effect of enhancing the electron transfer efficiency is achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a biological material with enhanced electron transfer efficiency and a preparation method and application thereof. Background technique [0002] With the development of society, people's demand for energy is increasing day by day, but resources such as oil and coal are non-renewable resources, and the large-scale combustion of oil and coal has brought environmental problems such as the greenhouse effect. Electric energy has the advantages of being renewable and clean. However, in the traditional power generation method, thermal power generation will burn a large amount of coal, which increases the cost of electricity and pollutes the environment; wind power generation and nuclear power generation are more location-dependent and cannot be deployed on a large scale. The delivery method is particularly important. [0003] Compared with other energy production methods, the ...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12N15/31C12N15/55C12N15/56H01M8/16C12R1/19
CPCC12N15/70C12N9/18C12N9/2402C12N9/2482C07K14/245C07K14/315C07K14/21C12Y301/01072C12Y302/01037C12Y302/01008H01M8/16Y02E50/10
Inventor 梁波杨建明杨京张鑫平
Owner QINGDAO AGRI UNIV
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