阅读说明：本技术 一种椰酵假单胞菌抑菌组合物及应用 (Coco ferment pseudomonas antibacterial composition and application thereof ) 是由 黄琼叶 杨亚琼 张亚宜 于 2021-07-13 设计创作，主要内容包括：本发明提供了一种椰酵假单胞菌抑菌组合物及应用,所述组合物包含(1)葡萄糖氧化酶；(2)葡萄糖或产葡萄糖组合物；所述产葡萄糖组合物是指能反应产生葡萄糖的组合物。本发明提供的椰酵假单胞菌抑菌组合物对于椰酵假单胞菌具有良好的抑制效果,将其用于食品加工中,对椰酵假单胞菌有较强抑菌作用,也可抑制椰酵假单胞菌产米酵菌酸毒素,且本发明组合物的成分用于食品加工能与食物原味有良好的适配性,长时间食用也不会给身体带来副作用。为防止食品椰酵假单胞菌污染提供了一种新型、天然、安全的方案。(The invention provides a pseudomonas cocofermentum bacteriostatic composition and application, wherein the composition comprises (1) glucose oxidase; (2) glucose or a glucose-producing composition; by glucose-producing composition is meant a composition that reacts to produce glucose. The pseudomonas cocofermentum antibacterial composition provided by the invention has a good inhibition effect on pseudomonas cocofermentum, has a strong antibacterial effect on pseudomonas cocofermentum when being used for food processing, and can also inhibit the pseudomonas cocofermentum from producing zymophyte toxin, and the components of the composition can have good adaptability to the original taste of food when being used for food processing, and can not bring side effects to the body after being eaten for a long time. Provides a novel, natural and safe scheme for preventing the food from being polluted by the food of the coconut fermentation pseudomonas.)

1. The pseudomonas natans bacteriostatic composition is characterized by comprising the following components (1) and (2);

(1) a glucose oxidase;

(2) a glucose and/or glucose-producing composition;

by glucose-producing composition is meant a composition that reacts to produce glucose.

2. The pseudomonas cocofermentum bacteriostatic composition according to claim 1, wherein the glucose-producing composition is selected from one or more of a starch-amylase composition, a cellulose-cellulase composition, a sucrose-sucrase composition or a maltose-maltase composition.

3. The pseudomonas cocofermentum bacteriostatic composition according to claim 1, wherein the mass ratio of the glucose oxidase to the glucose is 1: 0.1 to 20.

4. A preparation for inhibiting production of zymogenic acid by pseudomonas cocofermentum, which is characterized by comprising the pseudomonas cocofermentum bacteriostatic composition as described in any one of claims 1-3.

5. The use of the pseudomonas cocofermentum bacteriostatic composition as defined in any one of claims 1-3 in preparation of a pseudomonas cocofermentum bacteriostatic preparation.

6. The use of the pseudomonas cocofermentum bacteriostatic composition as defined in any one of claims 1-3 in inhibiting production of zymotic acid by pseudomonas cocofermentum.

7. Use according to claim 5 or 6, characterized in that it is a use in the food sector.

8. The use according to claim 7, wherein the pseudomonas cocoi bacteriostasis composition according to any one of claims 1 to 3 is mixed with a food raw material and processed to inhibit pseudomonas cocoi and/or inhibit production of zymogenic acid by pseudomonas cocoi in the food.

9. The use of claim 8, wherein the pseudomonas cocofermentum bacteriostatic composition is added into the food in an amount of 0.1-0.5 wt%.

10. Use according to claim 9, wherein the food comprises cereals, potatoes, agaric.

Technical Field

The invention belongs to the technical field of natural bacteriostats, and particularly relates to a pseudomonas cocoanut bacteriostasis composition and application thereof.

Background

The pseudomonas cocofermentum (pseudomonads cocovenenans) is a gram-negative bacillus brevis with two sections of blunt circles, no tooth spores and flagella, is widely distributed in the nature, and is facultative anaerobic and easy to grow on the surface of food. The fermentation starter Acid (Bongkrekic Acid) is a toxin which is produced by pseudomonas cocoanulata and can cause food poisoning, the fermentation starter Acid is unstable to Acid, oxidant and sunlight, but is stable to heat, and cannot be destroyed under the conditions of boiling at 100 ℃ and high pressure (121 ℃), so that cooked food polluted by the fermentation starter Acid can cause poisoning of people or animals, serious people can die, and common symptoms occur in stomach discomfort, nausea and vomiting, abdominal distension, abdominal pain and the like at the earliest. The vomiting was initially food or yellow-green watery, some coffee-like. After the symptoms of digestive tract, toxic hepatitis such as hepatomegaly and abnormal liver function may appear in clinical manifestations, and severe patients may have hepatic coma or even die.

In the prior art, research on a poisoning mechanism of pseudomonas cocoanut and preventive measures thereof is good, 2019 discloses that the pseudomonas cocoanut is distributed in the nature, and zymophyte toxins are easily produced in fermented corn flour products, deteriorated tremella and other deteriorated starch products (glutinous rice, millet, sorghum rice, potato powder and the like), so that the sales of rice flour and products thereof are seriously influenced, and huge loss is brought to manufacturers of the rice flour and the products. Effectively preventing and controlling the pollution of the pseudomonas cocofermentum and the generation of the zymotic acid, and has important significance for guaranteeing the food safety of China and maintaining the economic benefits of China.

In the prior art, the research on the influence of sugar on the toxicity generating capacity of the pseudomonas cocoanut, Wangjing, LiuxiuMei, 7 months in 1996, discloses that the toxicity generating capacity of the pseudomonas cocoanut is greatly improved by adding glucose and glycerol, and the toxicity generating amount is not lower than that of potato glucose agar. The toxicity of the pseudomonas natans is not greatly influenced by the sucrose and the starch, probably because the pseudomonas natans cannot decompose and utilize the sucrose and the starch. It is suggested that the toxic producing conditions of the bacteria should be avoided in the food processing process, and the conditions for inhibiting the toxic production are created, such as high salt (more than 1.5 percent), low pH (less than 5.5) and the like.

At present, the method is particularly important for controlling pseudomonas cocoanut in food and inhibiting the production of the fermentation bacteria acid, no specific antidote aiming at the fermentation bacteria acid is provided in the medical science, and the severity and the cure condition of the disease are only related to the ingested fermentation bacteria acid toxin.

Disclosure of Invention

The invention aims to overcome the defects of control of the pseudomonas cocofermentum in the prior art, and aims to provide the pseudomonas cocofermentum antibacterial composition and application.

And the glucose oxidase and the glucose ingredient are natural and safe, and the glucose oxidase and the glucose ingredient have good adaptability with the original taste of food when being used for food processing, and can not bring side effects to the body even if being eaten for a long time.

The invention aims to provide a pseudomonas natans bacteriostatic composition.

Another object of the present invention is to provide a preparation for inhibiting production of zymotic acid by Pseudomonas cococcensis comprising the above composition.

The invention further aims to provide application of the pseudomonas natans antibacterial composition in preparation of a pseudomonas natans antibacterial preparation.

The invention further aims to provide application of the pseudomonas cocoanut antibacterial composition in inhibiting production of the zymogenic acid by pseudomonas cocoanut.

The above purpose of the invention is realized by the following technical scheme:

the invention provides a pseudomonas cocofermentum bacteriostatic composition which comprises the following components (1) and (2);

(1) a glucose oxidase;

(2) a glucose and/or glucose-producing composition;

by glucose-producing composition is meant a composition that reacts to produce glucose.

In the composition, Glucose Oxidase (GOD) is distributed in animals, plants and microorganisms, is an important industrial enzyme in the food industry, is widely applied to aspects of deoxidation of foods such as wine, beer, fruit juice, milk powder and the like, flour improvement, food browning prevention and the like, and also has wide application in food rapid detection and biosensors. The research of the invention shows that the glucose oxidase and the glucose have good inhibition effect on the pseudomonas cococcensis after being mixed, therefore, the invention requests to protect the composition of the glucose oxidase and the glucose, and in addition, the glucose-based composition can be obtained by reacting various polysaccharides and oligosaccharides commonly used in the food industry with corresponding enzyme preparations thereof, and therefore, the combination of the composition which can react to generate the glucose and the glucose is also in the protection scope of the invention.

Further, the glucose-producing composition is selected from one or more of a starch-amylase composition, a cellulose-cellulase composition, a sucrose-sucrase composition or a maltose-maltase composition.

The components of the composition are natural and high in safety, and the enzyme can hydrolyze corresponding polysaccharide or oligosaccharide to generate glucose under the temperature condition of conventional food processing, and the generated glucose further plays an inhibiting role with glucose oxidase on pseudomonas cocoaccharensis.

Preferably, the mass ratio of the glucose oxidase to the glucose is 0.1-20.

Further preferably, the mass ratio of the glucose oxidase to the glucose is 1: 6-9.

Most preferably, the mass ratio of glucose oxidase to glucose is 1: 9.

based on further research results of the inventor, the composition can inhibit pseudomonas cocoanut and can also effectively inhibit the pseudomonas cocoanut to produce the zymotic acid. Therefore, the invention also claims a pseudomonas natans bacteriostatic preparation containing the pseudomonas natans bacteriostatic composition; and a preparation for inhibiting the production of the zymogenic acid by the pseudomonas cocoanut, which comprises the pseudomonas cocoanut bacteriostatic composition. The preparation comprises the coconut fermentation pseudomonas bacteriostatic composition as an effective component and also comprises acceptable auxiliary materials and/or processing aids in food.

In addition, the application of the pseudomonas cocofermentum antibacterial composition in preparing a pseudomonas cocofermentum antibacterial preparation and the application of the pseudomonas cocofermentum antibacterial composition in inhibiting the production of the zymotic acid by pseudomonas cocofermentum are also within the protection scope of the invention.

The composition of the invention can inhibit the pseudomonas cocofermentum or the product of the fermentation bacterium acid, and mainly controls and controls the bacteria and toxin in the food field. The pseudomonas cocoanut antibacterial composition is mixed with food raw materials for processing, and the pseudomonas cocoanut in the food raw materials can be effectively inhibited in the processing process; in the later storage process of the food, if the food is polluted by the pseudomonas cocofermentum due to environmental pollution or improper storage, the production of the zymotic acid by the pseudomonas cocofermentum in the food can be effectively inhibited.

Preferably, the addition amount of the pseudomonas cocofermentum bacteriostatic composition in the food is 0.1-0.5 wt%.

In general, there are 3 types of food contaminated by microorganisms in the environment that readily produce pseudomonas cocoi: (1) the cereal product is selected from fermented corn flour, glutinous corn dumpling flour, corn starch, fermented glutinous millet, glutinous rice cake, vinegar bean jelly, and rice noodles. (2) Deteriorated Tremella and Auricularia. (3) Potato products such as potato vermicelli, sweet potato noodles, sweet potato starch, etc. Therefore, the bacteriostatic composition can be used for the common food easily polluted by the pseudomonas cococcae.

Compared with the prior art, the invention has the beneficial effects that:

the first research of the invention shows that the pseudomonas cocoanut antibacterial composition has good inhibition effect on pseudomonas cocoanut, can effectively inhibit pseudomonas cocoanut and inhibit the pseudomonas cocoanut from producing zymophyte toxin when being used in food processing, has good adaptability to the original taste of food when being used for food processing, and does not bring side effect to human body after being eaten for a long time. Provides a novel, natural and safe scheme for preventing the food from being polluted by the food of the coconut fermentation pseudomonas.

Detailed Description

The present invention will be further described with reference to specific embodiments, but the present invention is not limited to the examples in any way. The starting reagents employed in the examples of the present invention are, unless otherwise specified, those that are conventionally purchased.

The pseudomonas cocoanulata is purchased from China culture resource bank collection center and is numbered BMZ 135745.

Example 1 Coco fermentation Pseudomonas bacteriostatic composition

This example provides 5 pseudomonas cocofermentum bacteriostatic compositions combined with glucose oxidase and glucose in different mass ratios:

(1) composition 1: the mass ratio of the glucose oxidase to the glucose is 1: 0.1;

(2) composition 2: the mass ratio of the glucose oxidase to the glucose is 1: 1;

(3) composition 3: the mass ratio of the glucose oxidase to the glucose is 1: 6;

(4) composition 4: the mass ratio of the glucose oxidase to the glucose is 1: 9;

(5) composition 5: the mass ratio of the glucose oxidase to the glucose is 1: 20.

example 2 Coco fermentation Pseudomonas bacteriostatic composition bacteriostatic test

1. Experimental methods

The inhibition effect of the pseudomonas cocofermentum antibacterial composition on pseudomonas cocofermentum is detected according to the national standard' determination of antibacterial activity of microbial source antibiotic secondary metabolites (GB/T38483-2020). The concentration of the Pseudomonas cocofermentum suspension in the test plate was set at 1X 10⁷CFU/mL. 5 Coco fermentation pseudomonas bacteria inhibiting compositions 1-5 of example 1 were added to the test plates, and the concentrations of the Coco fermentation pseudomonas bacteria inhibiting compositions in the test plates were adjusted to 3mg/mL, 1.5mg/mL, and 0.6mg/mL, respectively. After the test plate was cultured at 37 ℃ for 24 hours, the size of the zone of inhibition was observed.

The non-coconut fermentation pseudomonas bacteriostatic composition is used as a blank control group.

2. Results of the experiment

TABLE 1 inhibitory Effect of different Pseudomonas cococcharensis bacteriostatic compositions on Pseudomonas cococcharensis at different concentrations

As can be seen from the results in Table 1, compositions 1 to 4 had more significant effects on Pseudomonas cocoi than composition 5 at concentrations of 3mg/mL, 1.5mg/mL, 0.6 mg/mL.

Example 3 inhibition experiment of Pseudomonadaceae Coco bacteria bacteriostatic composition on Pseudomonadaceae Cocois and Miinomycelic acid in river powder

The composition based on glucose oxidase and glucose has obvious inhibition effect on pseudomonas cococcae, so the inventor further researches the inhibition effect of the composition based on glucose oxidase and glucose on pseudomonas cococcae and zymotic acid in starch food.

1. Experimental methods

(1) Grouping tests:

experimental groups: according to the conventional production process of the rice noodles: rice → washing → soaking → grinding of milk → steaming of pulp → cutting of noodles → cooling → finished product of rice noodles. After grinding, 0.3 wt% of composition 4 and 0.1mL/L of Pseudomonas cocoanut suspension (viable bacteria content 10) were added to the rice pulp before steaming⁷/mL), standing the rice milk overnight and then steaming the rice milk;

control group 1: the process of the same experimental group is different in that the composition 4 is not added in the processing process;

control group 2: the process is the same as the experimental group, and the difference is that 0.3 wt% of glucose oxidase is added into the rice pulp before the pulp steaming.

(2) And (3) respectively placing the fresh rice noodles prepared in the experimental group and the control group in a closed manner at the temperature of 0-10 ℃ for 15 days, and testing the contents of the pseudomonas cococcae and the fermentation acid in the rice noodles.

The detection method of the pseudomonas cocoanulata comprises the following steps: test of Pseudomonas cocoanut fermented Rice flour subspecies by national Standard of the people's republic of China (GB/T4789.29-2003).

The method for testing the content of the zymotic acid comprises the following steps: the determination of the food safety national standard of food for the fermentation of rice-ferment acid (GB 5009.189-2016).

2. Results of the experiment

TABLE 2 number of Pseudomonas cococcae in Hefen (CFU/g)

TABLE 3 content of Mimejic acid in the river powder (μ g/g)

As shown in tables 2 and 3, based on the further research results of the inventors, the pseudomonas cocoanut antibacterial composition provided by the invention is added to starch food such as rice noodles, so that the pseudomonas cocoanut can be effectively inhibited from generating zymophyte as well as being effectively inhibited from generating zymophytic acid by the pseudomonas cocoanut, wherein the rice noodles are polluted in the production or storage process. The effect of inhibiting pseudomonas cocoanut in the river powder by independently adding the glucose oxidase is not good, although the fermentation acid is not detected, the growth of the bacteria is very obvious, and the fermentation acid is inevitably generated along with the prolonging of the storage time.

Example 4 inhibition experiment of Pseudomonadaceae Cocois bacteriostatic composition on Pseudomonadaceae Cocois and Miermensic acid in Auricularia

The inventors further investigated the inhibitory effect of the combination of glucose oxidase and glucose on pseudomonas cococcae and mirermentans in agaric.

1. Experimental methods

(1) Grouping tests:

experimental groups: dividing dried northeast black fungus into three parts, respectively placing in three containers, adding 10 times of warm water, and adding Pseudomonas cocoanut suspension (viable bacteria content of 10) at a ratio of 0.1mL/L⁷/mL)。

Adding the composition 4 into a first container, wherein the adding amount is 0.1 wt%, uniformly stirring, and marking as an experimental group;

nothing was added to the second container and it was designated as control 1;

adding glucose oxidase into a third container, wherein the adding amount is 0.1 wt%, uniformly stirring, and marking as a control group 2;

standing and soaking for 3 hours after the operations are finished, fishing out and draining water, and putting into a fresh-keeping bag for sealed storage.

(2) And (3) respectively placing the agarics obtained from the experimental group and the control group in a closed manner at the temperature of 0-10 ℃ for 15 days, and testing the contents of the pseudomonas cocofermentum and the zymotic acid in the agarics.

The detection method of the pseudomonas cocoanulata comprises the following steps: test of Pseudomonas cocoanut fermented Rice flour subspecies by national Standard of the people's republic of China (GB/T4789.29-2003).

The method for testing the content of the zymotic acid comprises the following steps: the determination of the food safety national standard of food for the fermentation of rice-ferment acid (GB 5009.189-2016).

2. Results of the experiment

TABLE 4 Coco fermentation Pseudomonas count in Auricularia (CFU/g)

TABLE 5 content of Miermectinic acid in Auricularia (μ g/g)

As shown in tables 4 and 5, when the pseudomonas cocoanut bacteriostatic composition is added into agaric, not only can the pseudomonas cocoanut be effectively inhibited, but also the production of the zymotic acid by the pseudomonas cocoanut can be effectively inhibited. And the addition of glucose oxidase alone has no effect on the inhibition of the pseudomonas cocoanut in the agaric.

Example 5 inhibition of Pseudomonas cocoi bacteria bacteriostatic composition against Pseudomonas cocoi and Miermensic acid in Potato vermicelli

The inventors further investigated the inhibitory effect of the combination of glucose oxidase and glucose on pseudomonas cococcae and mirostrobin in potato vermicelli.

1. Experimental methods

(1) Grouping tests:

experimental groups: according to the conventional production process of the potato vermicelli: selecting material and extracting powder → mixing with starch → adding vitriol and flour → leaking strip in boiling water→ cold bath drying strip → bundling package. 1kg of potato flour was added to 5kg of water, followed by 0.5% by weight of composition 3 and 1mL of Pseudomonas cocoi suspension (viable bacteria content 10)⁷mL), after 12 hours of soaking, the water was filtered off and the starch was pressed dry for making vermicelli.

Control group 1: the difference from the experimental group was that composition 3 was not added;

control group 2: the difference from the experimental group was that 0.5 wt% glucose oxidase was added.

(2) And (3) respectively placing the potato vermicelli obtained from the experimental group and the potato vermicelli obtained from the control group in a closed manner at the temperature of 0-10 ℃ for 15 days, and testing the contents of the pseudomonas cococcae and the zymotic acid in the potato vermicelli.

The detection method of the pseudomonas cocoanulata comprises the following steps: test of Pseudomonas cocoanut fermented Rice flour subspecies by national Standard of the people's republic of China (GB/T4789.29-2003).

The method for testing the content of the zymotic acid comprises the following steps: the determination of the food safety national standard of food for the fermentation of rice-ferment acid (GB 5009.189-2016).

2. Results of the experiment

TABLE 6 number of Pseudomonas cococcae in Potato chips (CFU/g)

TABLE 7 content of Mimejic acid in Potato vermicelli (μ g/g)

As shown in tables 6 and 7, when the pseudomonas cocoanut bacteriostatic composition is added into the potato vermicelli, not only can the pseudomonas cocoanut be effectively inhibited, but also the pseudomonas cocoanut can be effectively inhibited from producing the zymotic acid. And the effect of inhibiting the pseudomonas cocoanut in the potato vermicelli by independently adding the glucose oxidase is poor.

Therefore, the pseudomonas cocoanut composition is used for common foods which are easily polluted by pseudomonas cocoanut, such as cereal foods, potato foods, agaric and the like, can effectively inhibit pseudomonas cocoanut in the foods, and can also effectively inhibit pseudomonas cocoanut from producing zymotic acid. The composition has good adaptability to the original taste of food when used in food processing, and has no adverse side effect after long-term consumption. Provides a novel, natural and safe scheme for preventing the food from being polluted by the food of the coconut fermentation pseudomonas.

It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.