CN103289907A - A candida utilis protein product and applications thereof in breeding industry - Google Patents
A candida utilis protein product and applications thereof in breeding industry Download PDFInfo
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- CN103289907A CN103289907A CN2013100878863A CN201310087886A CN103289907A CN 103289907 A CN103289907 A CN 103289907A CN 2013100878863 A CN2013100878863 A CN 2013100878863A CN 201310087886 A CN201310087886 A CN 201310087886A CN 103289907 A CN103289907 A CN 103289907A
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Abstract
The present invention relates to a solid fermentation preparation method for Candida utilis protein feed and applications of the feed in breeding industry. The Candida utilis is adopted as a fermentation strain, after slant culture and seed culture, the Candida utilis is inoculated in an optimized solid medium for culture at 25-28 DEG C, and large-scale fermentation is carried out after three-level amplification culture. The production process is simple and stable, the cell density and the protein content are high, and the fermentation product needs not separation and purification, so the Candida utilis protein feed is easy for popularization for use. Through adding the fermentation product of the Candida utilis into pig feed, good effects such as preventing gastrointestinal diseases, promoting growth, gaining weight and increasing feed conversion can be achieved in experiments, so the Candida utilis protein feed can be used as an animal feed additive to be applied in breeding industry.
Description
Technical field
The present invention relates to the microbial fermentation engineering field, specifically a kind of solid medium multiparity protein candiyeast engineering bacterium fermentation prepares the method for Candida utilis protein fodder and the application in aquaculture thereof.
Background technology
Candida utilis (Candida utilis) is named again and produces protein torula or edible torula, mycocandida Candida utilis kind, and cell is rounded, oval and cylindrical, and size is 3.5～4.5 μ m * 7～13 μ m.Liquid culture does not have the mould of pouncing on, and produces bacterial sediment and the thin epistasis of ring-type, can ferment.Bacterium colony on the malt extract medium is oyster white, and is level and smooth, glossy or matt, neat in edge or be mycelioid.On the Corn Meal Agar substratum with cover plate, only can generate some original pseudohyphas or underdeveloped pseudohypha, or not have mycelia.Suitable growth temperature is 25～28 ℃.It to the effect that animal produces is: can provide animal required part nutritive substance, and can regulate the animal intestinal microecological balance, improve feed digestibility, strengthen animal body immunizing power.The content of its protein and vitamins B is all than cereuisiae fermentum height, and it can be with urea and nitric acid as nitrogenous source, and not needing to add any somatomedin at substratum can grow.It can utilize five-carbon sugar and hexose, can utilize the sulfite waste liquor of paper industry, can also utilize molasses, wood saccharification liquid etc. to produce edible protein.
Candida utilis (Candida utilis) rich in proteins, carbohydrate and vitamin B group, protein content accounts for 45 ~ 55% of cell dry-matter, and lipid content is low, also contains multiple enzyme, gsh and other inorganic salt etc., and fermentation period is short, the zymotechnique maturation.The certification label that Candida utilis gives as a kind of GRAS(U.S. food medicine responsible institution) safe biologic, authenticated to can be used as the yeast of foodstuff additive by U.S. FDA (food and FAD), and can apply to food and pharmaceutical industry, be used for the production of products such as food protein and gsh.Some countries and regions such as Germany, the U.S. are used for feedstuff industry with Candida utilis widely, as good feed protein matter; Perhaps be used for the fermentation of materials such as stalk, rapeseed cake, be used for production or the detoxification of feed protein.It is the ability of organonitrogen that Candida utilis has stronger conversion inorganic nitrogen, and can produce great deal of bioactive substances in its fermenting process.Therefore, utilize Candida utilis production additive agent for feeding or starter to have a good application prospect.
Summary of the invention
The object of the present invention is to provide a kind of Candida utilis protein fodder fermentation method for producing.
Another object of the present invention is to provide this Candida utilis protein fodder as the application of additive in aquaculture.
The present invention is to use the Candida utilis bacterium of Guangdong Microbes Inst DSMZ as fermented bacterium, culture presevation numbering GIM2.148.By the Candida utilis protein fodder being carried out the research of culture medium prescription, pass through one-factor experiment, being defined as with Semen Maydis powder and wheat-flour is basestocks (mass ratio is 3:1), adds 2% glucose, and the suitableeest inorganic salt and somatomedin are respectively 0.5% K2HPO4 and 0.05% vitamin H; On this basis glucose, K2HPO4 and vitamin H have been carried out 3 factors, 3 horizontal quadratures tests, filtered out and optimize substratum and be: be basestocks (mass ratio is 3:1) with Semen Maydis powder and wheat-flour, glucose 2%, K2HPO4 1.0%, vitamin H 0.01%.
, add an amount of carbon source, nitrogenous source, inorganic salt and somatomedin and carry out the Candida utilis solid fermentation as basestocks with Semen Maydis powder and wheat-flour, reduce culture medium cost, improve Candida utilis nectar degree, for its application in aquaculture provides the basis.Candida utilis protein fodder through this method fermentative production contains materials such as rich in protein, vitamin B group, amino acid, is widely used as the protein supplements of animal-feed.It can promote growing of animal, shortens feeding period, increases meat amount and egg amount, and improvement meat and raising lean ratio improve the glossiness of fur, and can strengthen the resistance against diseases of cub fowl.
Embodiment
Embodiment 1 solid fermentation is produced the technical process of Candida utilis protein fodder
1. actication of culture:
The present invention adopt Candida utilis (
Candida utilis) as fermented bacterium, purchase in Guangdong Microbes Inst DSMZ, culture presevation numbering GIM2.148, with the Candida utilis bacterial classification of-80 ℃ of preservations at YPD solid (peptone 10g, yeast extract 5g, glucose 10g, agar powder 15g, add water to 1 liter, autoclave sterilization is standby) rule on the inclined-plane, cultivated 16 ~ 24 hours for 28 ℃; It and cereuisiae fermentum, kluyveromyces are the yeast as foodstuff additive by the U.S. FDA authentication, can apply to food, pharmacy and feed industry, comprise meat product industry.As Germany, the U.S. and Brazil, Candida utilis all is used as the feed of livestock in many countries.
2. preparation first order seed:
Candida utilis bacterial classification with activation, be inoculated in 200 mL YPD liquid (peptone 10g with transfering loop picking one ring, yeast extract 5g, glucose 10g adds water to 1 liter, autoclave sterilization is standby) in the substratum, triangular flask is placed shaking table, 28 ℃, 250rpm, after the shaking culture 20 hours, make primary seed solution.
3. prepare the solid fermentation substratum:
The solid butt: be basestocks (mass ratio is 3:1) with Semen Maydis powder and wheat-flour, glucose 2%, K
2HPO
41.0%, vitamin H 0.01%.It is the sterilized water of 1:1 ~ 2 that the solid butt is added mass ratio, and adjust pH to 5.5 stirs.Solid medium through 121 ℃, 20min autoclave sterilization, namely be can be used as the solid fermentation substratum after cooling and uses.
4. solid fermentation:
To insert the first order seed bacterium according to 8% inoculum size through the solid fermentation substratum of sterilising treatment, and stir, keeping humidity is 50 ~ 70%, and temperature is 25 ~ 28 ℃, cultivates about 16 hours, makes secondary seed.Secondary seed according to carrying out enlarged culturing in 8% the inoculum size adding sterilization solid fermentation substratum, is stirred, and keeping humidity is 50 ~ 70%, and temperature is 25 ~ 28 ℃, cultivates about 16 hours, makes three grades of seeds.Can be according to concrete cultivation consumption, further with three grades of seeds according to the further enlarged culturing of 8% inoculum size.The Candida utilis culture is carried out enumeration, and viable count reaches 1.5 * 10
9Cfu/g is bright bent.
5. the Candida utilis culture with solid fermentation carries out bake drying, pulverizes and packing, makes the additive of protein feed for livestock and poultry cultivation.
Embodiment 2 Candida utilis protein fodder products promote the application of child care pig growth
For the influence of checking Candida utilis culture to child care piglet growth performance and immunizing power, carried out culture experiment in certain child care workshop.
2.1 experimental design
Choose the weanling pig of 46 ages in days, totally 334, be divided into 2 processing at random according to body weight phase approximately principle, (wherein purebred piglet is 59 for 169 repetitions of control group, 110 of the assorted piglets of binary), 165 repetitions of experimental group (wherein purebred piglet is 57,108 of the assorted piglets of binary), every piglet is weighed separately, as a repetition.Handle (1) control group: do not add Candida utilis albumen in the basal diet, initial counterpoise is 9.11kg; (2) experimental group is added Candida utilis albumen 1.5% in the basal diet, initial counterpoise is 9.44 kg.Trial period is 35d, is divided into preliminary trial period 4d and is just trying phase 31d.Experiment lasts 35 days (branchs 4 days preliminary trial periods, just trying 31 days phases), and experimental data is initial with 50d, and 81d finishes, and adds the time of Candida utilis protein fodder from 50d to 81d, amounts to interpolation 31 days.
2.2 experiment daily ration
Corn-soybean meal diet is adopted in experiment, for preparing burden certainly at the testing ground.
2.3 feeding and management
Experiment is carried out on the pig farm, Hebei, and feeding and management is carried out according to former feeding and management rules.
2.4 detection method and index
Weigh and ramming material the next day in each experiment day, calculate average daily gain (average daily weight, ADG), average daily ingestion amount (average daily feed intake, ADFI) and feedstuff-meat ratio (Feed:Gain).
Average daily gain (Kg/ days)=total augment weight sum (Kg)/experiment fate
Average daily ingestion amount (the Kg/ previous day)=total feed consumption rate (Kg)/all survival pig's heads are counted sum (previous day)
Consume the gain in weight of forage volume/contemporaneity body weight feedstuff-meat ratio=a certain period
2.5 statistical study
Be infrastest unit with every pig, indexs such as average initial weight, average end weight, average weight gain, average daily gain adopt the SPSS softwares, compare difference (p＜0.01) between two samples carrying out the t-check analysis under the oneway ANOVA.
2.6 result and discussion
As can be seen from Table 2, experimental group is compared dead survival rate and has been improved 1.76 percentage points with control group, and incubation rate has improved 7.06 percentage points, shows that the health level of swinery and resistance against diseases are better than control group; Average weight gain has improved 3.92kg(p＜0.01); Average daily gain has significantly improved 126.3g(p＜0.01); Feedstuff-meat ratio has descended 0.27, weighs the above pig's head of 15 kg and 20kg from the end and counts proportion as can be seen, and the ratio of experimental group is obviously greater than control group.
The analysis of table 2 production performance
In the table after the data * represent t-check analysis result,
pUnder＜0.01 level, * * represents that difference is extremely remarkable
In addition, feeding and management every day is observed, and the swinery fur was ruddy, energetic after experimental group was fed 7 days, the food consumption height; Hair color after 20 days, vigor effect are more obvious; It is slender that experiment in 31 days finishes a back pig build, and skeleton elongates.Above result shows: Candida utilis albumen can improve the production performance of child care pig effectively, can improve day weight gain, surviving rate and the feed conversion rate of child care piglet, reduces feedstuff-meat ratio, realizes the nonreactive green cultivation.
Claims (7)
1. the preparation method of a Candida utilis protein formulation may further comprise the steps:
(1) actication of culture:
The present invention adopts Candida utilis (Candida utilis) as fermented bacterium, with the Candida utilis bacterial classification of-80 ℃ of preservations at YPD solid (peptone 10g, yeast extract 5g, glucose 10g, agar powder 15g, add water to 1 liter, autoclave sterilization is standby) rule on the inclined-plane, cultivated 16 ~ 24 hours for 28 ℃;
(2) preparation first order seed:
Candida utilis bacterial classification with activation, be inoculated in 200 mL YPD liquid nutrient medium (peptone 10g with transfering loop picking one ring, yeast extract 5g, glucose 10g adds water to 1 liter, autoclave sterilization is standby) in, triangular flask is placed shaking table, 28 ℃, 250rpm, after the shaking culture 16 ~ 24 hours, make primary seed solution;
(3) preparation solid fermentation substratum: solid butt: be basestocks (mass ratio is 3:1) with Semen Maydis powder and wheat-flour, glucose 2%, K2HPO4 1.0%, vitamin H 0.01%; It is the sterilized water of 1:1 ~ 2 that the solid butt is added mass ratio, and adjust pH to 5.5 stirs; Solid medium through 121 ℃, 20min autoclave sterilization, namely be can be used as the solid fermentation substratum after cooling and uses;
(4) solid fermentation: will insert the first order seed bacterium according to 8 ~ 10% inoculum size through the solid fermentation substratum of sterilising treatment, and stir, keeping humidity is 50 ~ 70%, and temperature is 25 ~ 28 ℃, cultivates about 16 hours, makes secondary seed; Secondary seed according to carrying out enlarged culturing in 8 ~ 10% the inoculum size adding sterilization solid fermentation substratum, is stirred, and keeping humidity is 50 ~ 70%, and temperature is 25 ~ 28 ℃, cultivates about 16 hours, makes three grades of seeds; Can be according to concrete cultivation consumption, further with three grades of seeds according to the further enlarged culturing of 8 ~ 10% inoculum size.
2. Candida utilis egg white solid fermentative production according to claim 1 is characterized in that described solid butt: be basestocks (mass ratio is 3:1) with Semen Maydis powder and wheat-flour, and glucose 2%, K2HPO4 1.0%, vitamin H 0.01%.
3. Candida utilis egg white solid fermentation according to claim 1, it is characterized in that: culture temperature is 25 ~ 28 ℃, humidity is 50 ~ 70%.
4. Candida utilis egg white solid according to claim 1 fermentation is characterized in that: at different levels kinds of daughter bacteria inoculum sizes are 8 ~ 10% inoculum size.
5. Candida utilis egg white solid fermentation according to claim 1, it is characterized in that: used bacterial classification is the Candida utilis kind of mycocandida.
6. Candida utilis egg white solid fermentation according to claim 1, it is characterized in that: the addition as fodder additives is 0.5 ~ 2%.
7. Candida utilis egg white solid fermentation according to claim 1, it is characterized in that: the Application Areas as fodder additives is additive for farm animal feed.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104206704A (en) * | 2014-05-27 | 2014-12-17 | 浙江科峰生物技术有限公司 | Preparation method of yeast culture |
| CN104522417A (en) * | 2014-12-30 | 2015-04-22 | 青岛农业大学 | Micro-granular feed for larva fish of bostrichthys sinensis and preparation method of micro-granular feed |
| CN105441344A (en) * | 2015-12-16 | 2016-03-30 | 山东宝来利来生物工程股份有限公司 | Candida utilis having antioxidant function and application thereof |
| CN107641602A (en) * | 2017-07-13 | 2018-01-30 | 常州市萤火虫生物科技有限公司 | One plant of candida utili and its fermentation lay eggs it is white in application |
| CN112175846A (en) * | 2020-10-22 | 2021-01-05 | 河南省科学院生物研究所有限责任公司 | Candida utilis strain UCY-11 and application thereof in preparation of fermented hybrid broussonetia papyrifera feed |
| CN114836331A (en) * | 2022-04-26 | 2022-08-02 | 内蒙古工业大学 | Candida utilis and application thereof |
| CN115011491A (en) * | 2022-07-06 | 2022-09-06 | 厦门大昌生物技术服务有限公司 | Culture process of candida utilis |
| CN116121085A (en) * | 2022-09-14 | 2023-05-16 | 大连理工大学 | Cold-resistant yeast suitable for low-temperature aquaculture and biocontrol application thereof |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104206704A (en) * | 2014-05-27 | 2014-12-17 | 浙江科峰生物技术有限公司 | Preparation method of yeast culture |
| CN104522417A (en) * | 2014-12-30 | 2015-04-22 | 青岛农业大学 | Micro-granular feed for larva fish of bostrichthys sinensis and preparation method of micro-granular feed |
| CN105441344A (en) * | 2015-12-16 | 2016-03-30 | 山东宝来利来生物工程股份有限公司 | Candida utilis having antioxidant function and application thereof |
| CN105441344B (en) * | 2015-12-16 | 2018-11-09 | 山东宝来利来生物工程股份有限公司 | One plant of candida utili and its application with anti-oxidation function |
| CN107641602A (en) * | 2017-07-13 | 2018-01-30 | 常州市萤火虫生物科技有限公司 | One plant of candida utili and its fermentation lay eggs it is white in application |
| CN107641602B (en) * | 2017-07-13 | 2020-09-29 | 江苏黑钻生物工程有限公司 | Candida utilis and application thereof in protein production through fermentation |
| CN112175846A (en) * | 2020-10-22 | 2021-01-05 | 河南省科学院生物研究所有限责任公司 | Candida utilis strain UCY-11 and application thereof in preparation of fermented hybrid broussonetia papyrifera feed |
| CN112175846B (en) * | 2020-10-22 | 2022-06-17 | 河南省科学院生物研究所有限责任公司 | Candida utilis strain UCY-11 and application thereof in preparation of fermented hybrid broussonetia papyrifera feed |
| CN114836331A (en) * | 2022-04-26 | 2022-08-02 | 内蒙古工业大学 | Candida utilis and application thereof |
| CN114836331B (en) * | 2022-04-26 | 2023-04-25 | 内蒙古工业大学 | Candida utilis and application thereof |
| CN115011491A (en) * | 2022-07-06 | 2022-09-06 | 厦门大昌生物技术服务有限公司 | Culture process of candida utilis |
| CN116121085A (en) * | 2022-09-14 | 2023-05-16 | 大连理工大学 | Cold-resistant yeast suitable for low-temperature aquaculture and biocontrol application thereof |
| CN116121085B (en) * | 2022-09-14 | 2024-05-07 | 大连理工大学 | Cold-resistant yeast suitable for low-temperature aquaculture and biocontrol application thereof |
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Application publication date: 20130911 |