Microorganism having detoxifizyme activity and with fermentation residues applicable to feed

A technology for detoxifying enzymes and recombining microorganisms, applied in the field of microorganisms, which can solve problems such as waste

Pending Publication Date: 2016-06-29
SHANGHAI RES & DEV CENT OF INDAL BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If these fermentation wastes rich in yeast cell components

Method used

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  • Microorganism having detoxifizyme activity and with fermentation residues applicable to feed
  • Microorganism having detoxifizyme activity and with fermentation residues applicable to feed
  • Microorganism having detoxifizyme activity and with fermentation residues applicable to feed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Construction of a yeast strain derived from aflatoxin detoxification enzyme from Peniophorasp.

[0069] According to the reported detoxification enzyme sequence from Peniophora sp. aflatoxin (International Journal of Food Microbiology 135 (2009) 47-52), the complete ORF sequence (Genbank Y18012) encoding aflatoxin detoxification enzyme was synthesized by the method of whole gene synthesis.

[0070] Using the Gibsonassembly method, the aflatoxin degradation gene was ligated into the pCpA1 / G-XI vector (Diao et al. BMC Biotechnology 2013, 13:110) and replaced with the XI gene in the pCpA1 / G-XI vector, and the obtained plasmid was named pCpA1-HQ1. The sequence was confirmed to be correct by sequencing. The aflatoxin detoxification enzyme gene is located between the TPI1 promoter and the CYC1 terminator.

[0071] Transform pCpA1-HQ1 into Saccharomyces cerevisiae CCTCCM94055 (CCTCC is China Center for Type Culture Collection), so as to obtain dozens of yeast strain transform...

Embodiment 2

[0074] Yeast with aflatoxin detoxifying enzyme activity of Peniophorasp. can ferment (enzyme change) grain into ethanol

[0075] The five transformants HQ11, HQ12, HQ13, HQ14, HQ15 constructed in Example 1 and the control strain CCTCCM94055 were used for ethanol fermentation respectively.

[0076] The raw materials with no aflatoxin detected were prepared according to the literature (Research on the Production of Ethanol by Fermentation of Corn Raw Meal, Food and Fermentation Industry, Issue 02, 2008), and the shake flask test of fermentation to produce ethanol was carried out. Before the start of fermentation, 10 mg / L aflatoxin was added to the medium. After 96 hours of fermentation, the ethanol output of each strain was basically the same, and the utilization rate of starch was between 85% and 90%.

[0077] The content of aflatoxin B1 was detected before and after fermentation. As shown in Table 1, the aflatoxin in the control strain group only decreased by 7.2%, which may...

Embodiment 3

[0083] Aflatoxin detoxification enzyme activity of residual bacteria after fermentation

[0084] Take 10 mg of wet yeast cells obtained by fermenting with moldy raw materials in Example 2, and resuspend in 800 μL of PBS buffer. Take 200μL aflatoxin B1 (500ppm) for reaction.

[0085] The blank control group was 800 μL PBS buffer. Take 200μL aflatoxin B1 PBS solution (500ppm) for reaction.

[0086] After each treatment was reacted at 35 degrees for 5 hours, the concentration of aflatoxin was determined.

[0087] As can be seen from Table 2, after the treatment of each bacterial strain, aflatoxins were all reduced. But the control strain had the smallest reduction, probably only by physical adsorption. When the yeast cell with aflatoxin detoxification enzyme gene was treated with aflatoxin, the reduction of aflatoxin was higher than that of the control strain. It shows that the aflatoxin detoxification enzyme in the bacteria has played a role.

[0088] The effect of table 2 t...

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Abstract

The invention discloses a microorganism having a detoxifizyme activity and with fermentation residues applicable to feed. When a recombinant microorganism disclosed by the invention is fermented by taking mildewed grains or grains containing mycotoxins as fermentation raw materials, a starch utilization rate is at least 75%, and the recombinant microorganism contains exogenous mycotoxin detoxifizyme gene expression cassettes, wherein mycotoxin detoxifizyme is selected from the following detoxifizymes: aflatoxin detoxifizyme, zearalenone detoxifizyme, vomitoxin detoxifizyme, T-2 toxin detoxifizyme or a combination thereof; and in addition, compared with a wild strain or an original strain which is free from the exogenous mycotoxin detoxifizyme gene expression cassettes, the recombinant microorganism can increase a detoxifying activity for at least one mycotoxin by an increased range [delta] T which is more than or equal to 50%. According to the microorganism disclosed by the invention, a substrate using range is widened and cost of raw materials is reduced; an existing fermentation technology is adopted in the whole course of a degrading process and the addition of additional cost is avoided; and in addition, the value of a fermentation product can be also increased.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a microorganism containing detoxification enzyme activity and fermentation residues that can be used for feed. Background technique [0002] Mycotoxins are an important threat to the safety of livestock and poultry feed and human food worldwide. It is estimated that about 25% of the world's grain is contaminated with mycotoxins every year. Mycotoxins are naturally occurring by-products of the growth of molds and are highly toxic. Killing the mold only stops the production of mycotoxins, but does not eliminate the mycotoxins already present. Mycotoxins pose a series of problems that cannot be ignored in the food industry, feed industry and animal husbandry. A large amount of contaminated food is inedible, causing huge waste; after animals eat mildewed feed containing mycotoxins, not only may chronic or acute symptoms of poisoning, increased mortality and reduced feed conversion ra...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N1/19A23K1/14A23K1/16C12R1/865C12R1/145
Inventor 杨晟杨俊杰蒋宇陶荣盛
Owner SHANGHAI RES & DEV CENT OF INDAL BIOTECH
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