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Method of using surfactant to increase yield of Hb polypeptide prepared through degradation of hemoglobin by aspergillus niger

A surfactant and hemoglobin technology, applied in the direction of hemoglobin/myoglobin, microbial-based methods, biochemical equipment and methods, etc., to achieve cost saving, inhibit microbial absorption of polypeptide products, and simple operation

Inactive Publication Date: 2016-08-17
SERICULTURE & AGRI FOOD RES INST GUANGDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of Aspergillus niger to degrade hemoglobin (Hb) can save a lot of cost and enzyme costs, and increase the yield of Hb polypeptides, which is of great significance for the production of functional polypeptides, but no one has yet conducted research on this aspect

Method used

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  • Method of using surfactant to increase yield of Hb polypeptide prepared through degradation of hemoglobin by aspergillus niger
  • Method of using surfactant to increase yield of Hb polypeptide prepared through degradation of hemoglobin by aspergillus niger
  • Method of using surfactant to increase yield of Hb polypeptide prepared through degradation of hemoglobin by aspergillus niger

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Such as figure 1 As shown in, the method that utilizes tensio-active agent to improve Aspergillus niger to degrade hemoglobin to prepare Hb polypeptide yield provided by this example comprises the following steps:

[0041] (1) Preparation of Hb powder: add sodium citrate 5g / L (for anticoagulation treatment) to the storage tank receiving fresh chicken blood, centrifuge at 4°C and 12000r / min for 15min after collection, discard the supernatant to obtain the lower layer The precipitate was washed twice with an equal volume of normal saline, and then an equal volume of ultrapure water was added to swell and break the wall for 10 minutes in an ultrasonic instrument (power 3000W), and placed for 10 minutes, then centrifuged at 4°C and 6000r / min for 15 minutes, and the precipitate was discarded. The Hb supernatant was vacuum freeze-dried to obtain Hb powder;

[0042] (2) Preparation of Aspergillus niger seed solution: select Aspergillus niger (available from China Ordinary Mi...

Embodiment 2

[0050] Such as figure 1 As shown in, this example provides the method that utilizes food-grade surfactant to improve Aspergillus niger degradation hemoglobin to prepare the Hb polypeptide yield, comprising the following steps:

[0051] (1) Preparation of Hb powder: add sodium citrate 5g / L to the storage tank receiving fresh chicken blood, centrifuge at 4°C and 12000r / min for 15min after collection, discard the supernatant, and obtain the lower sediment plus an equal volume of physiological Wash with salt water twice, add an equal volume of ultrapure water, swell and break the wall in an ultrasonic instrument for 10 minutes, and place it for 10 minutes. Then centrifuge at 4°C and 6000r / min for 15min, discard the precipitate to obtain the Hb supernatant, and vacuum freeze-dry to obtain the Hb powder;

[0052] (2) Preparation of Aspergillus niger seed solution: select Aspergillus niger (available from China Ordinary Microorganism Collection Center, Guangdong Provincial Microbio...

Embodiment 3

[0060] Such as figure 1 Shown in, the method that utilizes food-grade surfactant provided by this example to improve Aspergillus niger degradation hemoglobin to prepare Hb polypeptide yield, comprises the following steps:

[0061] Steps (1)-(2) are the same as in Example 1.

[0062] (3) Addition of food-grade surfactant: Add the prepared seed liquid to the above fermentation medium at an inoculum amount of 9% (v / v) to ferment for 36 hours, and put it on a ultra-clean bench for ultraviolet sterilization with a pipette gun. Add 50% dodecyl glucoside for 30 minutes to the fermentation medium with a final concentration of 8g / L, shake and mix to continue fermentation for 16 hours, and cultivate at a shaking speed of 150rpm and a temperature of 30°C.

[0063] The slant medium, seed medium and fermentation medium are the same as in Example 1.

[0064] (4) Detection of Hb polypeptide yield: After the fermentation is completed, the fermentation supernatant is obtained by filtration ...

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Abstract

The invention discloses a method of using surfactant to increase yield of Hb polypeptide prepared through degradation of hemoglobin by aspergillus niger. The method comprises the following steps: (1) preparation of Hb powder; (2) preparation of an aspergillus niger seed liquid; (3) preparation of hemoglobin Hb polypeptide. Specifically, the aspergillus niger seed liquid is inoculated into fermentation medium containing Hb powder for fermentation; surfactant is added in the process of fermentation; after even mixing, fermentation continues until completion; subsequent treatment is required to obtain Hb polypeptide. The method is characterized in that chicken blood hemoglobin (Hb) is used as fermentation substrate, and food grade surfactant is added to increase the yield of Hb polypeptide prepared through the degradation of hemoglobin by aspergillus niger. Therefore, chicken blood is recovered; environmental protection is ensured; the utilization rate of aspergillus niger and the yield of Hb polypeptide are increased; the cost is reduced.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for improving the yield of Hb polypeptide prepared by aspergillus niger by degrading hemoglobin by using a surfactant. Background technique [0002] According to the statistics of the United States Department of Agriculture, the global chicken production in 2014 was about 85.292 million tons. The annual output of chicken meat in China ranks second in the world, with an annual output of 12.7 million tons. When a large number of live chickens are slaughtered, a large number of by-products - chicken blood will be produced. Due to the characteristics of chicken blood, such as strong smell, bright color, poor palatability, indigestibility, etc., coupled with the difficulty of collecting and storing chicken blood, it leads to Most of them are discharged as waste, which not only wastes precious resources, but also pollutes the environment. Chicken b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C12R1/685
CPCC12P21/06C07K14/805
Inventor 刘学铭张炫程镜蓉陈智毅张友胜张业辉
Owner SERICULTURE & AGRI FOOD RES INST GUANGDONG ACAD OF AGRI SCI
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