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Artificial synthetic material with biological activity and agricultural application thereof

A synthetic and artificial technology, applied in the fields of chemicals for biological control, agriculture, application, etc., can solve the problems of using hydrophilic protein and/or hemoglobin, etc., to achieve precise and efficient fertilization, directional binding and accurate root surface Efficient, wide-ranging effects

Pending Publication Date: 2022-01-07
北京绿氮生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In the prior art, there are only reports of PHA used as a seed coating alone, but there is no report on the use of hydrophilic proteins and / or hemoglobin

Method used

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  • Artificial synthetic material with biological activity and agricultural application thereof
  • Artificial synthetic material with biological activity and agricultural application thereof
  • Artificial synthetic material with biological activity and agricultural application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Induced Expression and Purification of Artificial Hydrophilic Protein in Escherichia coli

[0049] experiment procedure:

[0050] (1) IPTG induces recombinant protein expression

[0051] (1) Streak activation of the successfully constructed recombinant strain BL-DosHM. Pick a single colony and place it in 3 mL of fresh LB medium (containing kanamycin 50 mg / mL), and cultivate overnight at 37° C. and 220 rpm with shaking.

[0052] (2) The next day, the seed solution was transferred to 20mL LB medium containing Km (50mg / mL) at a ratio of 1:100, and cultured with shaking at 37°C and 220rmp, with an OD600 of about 0.6-0.8.

[0053] (3) Adding IPTG at a final concentration of 0.5 mM and overnight at 16° C. (at the same time, no IPTG was added as a blank control) to induce Escherichia coli to perform exogenous expression.

[0054] (4) Take the induced bacterial liquid, centrifuge at 5,000rmp for 5min, and collect the bacterial cells. The collected cells were shak...

Embodiment 2

[0065] Example 2 Induced expression and purification of artificial hemoglobin in Escherichia coli

[0066] experiment procedure:

[0067] (1) IPTG induces recombinant protein expression

[0068] (1) Streak activation of the successfully constructed recombinant strain BL-SYMB. Pick a single colony and place it in 3 mL of fresh LB medium (containing kanamycin 50 mg / mL), and cultivate overnight at 37° C. and 220 rpm with shaking.

[0069] (2) The next day, the seed solution was transferred to 20mL LB medium containing Km (50mg / mL) at a ratio of 1:100, and cultured with shaking at 37°C and 220rmp, with an OD600 of about 0.6-0.8.

[0070] (3) Adding IPTG with a final concentration of 0.5 mM and culturing at 20° C. for 10 hours to induce E. coli to perform exogenous expression.

[0071] (4) The induced bacterial solution was centrifuged at 5,000 rpm for 5 minutes to collect the bacterial cells. The collected cells were shaken and suspended with NTA-0 buffer (1 / 20 culture volume)...

Embodiment 3

[0082] Embodiment 3 has the preparation of the synthetic material of biological activity

[0083] experiment procedure:

[0084] (1) The composition of raw materials is as follows:

[0085] Short-chain PHA is a commercial product, purchased from Shenzhen Ecoman Biotechnology Co., Ltd.; artificial hydrophilic protein was dissolved in 50mM Tris-HCl buffer at a concentration of 0.5g / ml; artificial hemoglobin was dissolved in 50mM Tris-HCl buffer In, the concentration is 0.5g / ml.

[0086] (2) According to the above-mentioned raw materials, it is prepared by the following steps ( image 3 ):

[0087] (1) Sonicate an appropriate amount of short-chain PHA aqueous solution with an ultrasonic instrument between 20kHz-100kHz for 5min to form the first solution;

[0088] (2) Mix artificial hydrophilic protein and artificial hemoglobin dissolved in 50mM Tris-HCl buffer at a ratio of 1:1, and stir for 15min to form a second solution;

[0089] (3) The first solution after sonication is...

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Abstract

The invention provides an artificial synthetic material with biological activity and is applied to agriculture. The artificial synthetic material contains artificial hydrophilic protein, artificial hemoglobin and polyhydroxyalkanoate (PHA) which are designed and synthesized in the invention. The material can be used for preparing micron-sized active artificial micro-capsules, wrapping agricultural microbial excellent strains with the characteristics of nitrogen fixation, phosphorus and potassium solubilization, disease and pest resistance, growth promotion and the like to form a novel bacterial fertilizer, and can also be used in the agricultural fields of preparation of novel crop seed active coating agents and the like. The material has the unique properties of keeping thallus activity, directionally combining with root surface, accurately and efficiently fertilizing and the like, and is suitable for modern agricultural production modes such as mechanical seeding, water and fertilizer integrated drip irrigation, soilless culture and the like.

Description

Technical field: [0001] The invention relates to an artificial synthetic material with biological activity and its application in agriculture. Background technique: [0002] Ideal agricultural microbial products (preparations) should have excellent characteristics such as plasticity, degradability, water retention, root-surface affinity, and oxygen and carbon supply, so as to achieve nitrogen fixation, phosphorus and potassium decomposition, and resistance to diseases and insect pests to promote plant growth. . [0003] At present, microbial fertilizer products have problems such as poor strain viability, short shelf life caused by low activity, single dosage form, difficulty in long-distance transportation, complicated application methods, and unstable effects. In addition, products such as nitrogen-fixing microbial agents are greatly affected by stress factors such as rhizosphere ammonia nitrogen and adversity, resulting in low nitrogen fixation efficiency in the field, u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/805C07K14/195C12N15/12C12N15/31A01N47/44A01P21/00C05F11/08C05G3/00C05G3/60C05G3/80
CPCC07K14/805C07K14/195A01N47/44C05F11/08C05G3/00C05G3/60C05G3/80A01P21/00
Inventor 林敏柯秀彬燕永亮王劲战嵛华周正富
Owner 北京绿氮生物科技有限公司
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